pCDH-EF1-Luc2-T2A-tdTomato

pCDH-EF1-Luc2-T2A-tdTomato

载体基本信息

出品公司:	Ybscience
载体名称:	pCDH-EF1-Luc2-T2A-tdTomato
质粒类型:	哺乳动物细胞载体；慢病毒包装载体
高拷贝/低拷贝:	高拷贝
克隆方法:	限制性内切酶，多克隆位点
启动子:	EF1 alpha
载体大小:	9.5 kb
5' 测序引物及序列:	EF1引物：CTCCACGCTTTGCCTGACCCTGCTT
3' 测序引物及序列:	--
载体标签:	C-Luciferase
载体抗性:	氨苄青霉素
筛选标记:	--
克隆菌株:	DH5α 等
宿主细胞（系）:	常规细胞系（293、CV-1、CHO等）
备注:	包装质粒是pLP1、pLP2、pLP/VSVG。
稳定性:	稳表达
组成型/诱导型:	组成型
病毒/非病毒:	慢病毒

 

载体质粒图谱和多克隆位点信息

载体简介

 

载体序列

LOCUS       pCDH-EF1-Luc2-T2A-tdTomato                9565 bp ds-DNA     circular SYN 30-AUG-2016
DEFINITION  pCDH-EF1-Luc2-T2A-tdTomato vector
COMMENT     This file is created by Vector NTI
            http://www.biofeng.com/
COMMENT     ORIGDB|GenBank
COMMENT     VNTAUTHORNAME|biofeng.com|
FEATURES             Location/Qualifiers
     source          1..9565
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(10..26)
                     /note="M13 fwd"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        500..604
                     /gene="bla"
                     /note="AmpR promoter"
     CDS             605..1465
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
     rep_origin      1636..2224
                     /direction=RIGHT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    2512..2533
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding site"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2548..2578
                     /note="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    2586..2602
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="lac operator"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     2610..2626
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      2664..2799
                     /note="SV40 ori"
                     /note="SV40 origin of replication"
     polyA_signal    2818..2939
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     LTR             3011..3244
                     /note="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     misc_feature    3318..3906
                     /note="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             3424..3435
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /note="Factor Xa site"
                     /translation="IEGR"
     CDS             complement(3913..5343)
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /note="tdTomato"
                     /note="mammalian codon-optimized"
     CDS             complement(5353..5406)
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /note="T2A"
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             complement(5419..7068)
                     /codon_start=1
                     /product="firefly luciferase"
                     /note="luciferase"
                     /note="synthetic luc2 version of the luciferase gene"
     LTR             7108..7376
                     /note="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     promoter        complement(7389..7600)
                     /note="EF-1-alpha core promoter"
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     misc_feature    7645..7762
                     /note="cPPT/CTS"
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    8257..8490
                     /note="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    8983..9108
                     /note="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             9152..9332
                     /note="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
ORIGIN
        1 cagcttggca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca
       61 acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg
      121 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggcgcc tgatgcggta
      181 ttttctcctt acgcatctgt gcggtatttc acaccgcata tggtgcactc tcagtacaat
      241 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc
      301 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
      361 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt
      421 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
      481 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
      541 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
      601 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
      661 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
      721 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
      781 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
      841 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
      901 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
      961 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     1021 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     1081 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     1141 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     1201 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     1261 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     1321 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     1381 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     1441 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     1501 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     1561 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     1621 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     1681 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     1741 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta
     1801 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     1861 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     1921 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     1981 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     2041 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     2101 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     2161 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     2221 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     2281 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     2341 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     2401 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
     2461 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
     2521 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
     2581 tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg attacgaatt
     2641 tgggccgtct ctgcaaaagt ctaggcctcc aaaaaagcct cctcactact tctggaatag
     2701 ctcagaggcc gaggcggcct cggcctctgc ataaataaaa aaaattagtc agccatgggg
     2761 cggagaatgg gcggaactgg gcggagttag gggcgggata gctagagcca gacatgataa
     2821 gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt
     2881 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagttc
     2941 ctctcactct ctgatattca tttctttgca agttataaat actgaataat aagatgacat
     3001 gaactactac tgctagagat tttccacact gactaaaagg gtctgaggga tctctagtta
     3061 ccagagtcac acaacagacg ggcacacact acttgaagca ctcaaggcaa gctttattga
     3121 ggcttaagca gtgggttccc tagttagcca gagagctccc aggctcagat ctggtctaac
     3181 cagagagacc cagtacaagc aaaaagcaga tcttattttc gttgggagtg aattagccct
     3241 tccagtcccc ccttttcttt taaaaagtgg ctaagatcta cagctgcctt gtaagtcatt
     3301 ggtcttaaag gtaccaggcg gggaggcggc ccaaagggag atccgactcg tctgagggcg
     3361 aaggcgaaga cgcggaagag gccgcagagc cggcagcagg ccgcgggaag gaaggtccgc
     3421 tggattgagg gccgaaggga cgtagcagaa ggacgtcccg cgcagaatcc aggtggcaac
     3481 acaggcgagc agccaaggaa aggacgatga tttccccgac aacaccacgg aattgtcagt
     3541 gcccaacagc cgagcccctg tccagcagcg ggcaaggcag gcggcgatga gttccgccgt
     3601 ggcaataggg agggggaaag cgaaagtccc ggaaaggagc tgacaggtgg tggcaatgcc
     3661 ccaaccagtg ggggttgcgt cagcaaacac agtgcacacc acgccacgtt gcctgacaac
     3721 gggccacaac tcctcataaa gagacagcaa ccaggattta tacaaggagg agaaaatgaa
     3781 agccatacgg gaagcaatag catgatacaa aggcattaaa gcagcgtatc cacatagcgt
     3841 aaaaggagca acatagttaa gaataccagt caatctttca caaattttgt aatccagagg
     3901 ttgattgtcg acttacttgt acagctcgtc catgccgtac aggaacaggt ggtggcggcc
     3961 ctcggagcgc tcgtactgtt ccacgatggt gtagtcctcg ttgtgggagg tgatgtccag
     4021 cttggtgtcc acgtagtagt agccgggcag ttgcacgggc ttcttggcca tgtagatggt
     4081 cttgaactcc accaggtagt ggccgccgtc cttcagcttc agggcctggt ggatctcgcc
     4141 cttcagcacg ccgtcgcggg ggtacaggcg ctcggtggag gcctcccagc ccatggtctt
     4201 cttctgcatt acggggccgt cgggggggaa gttggtgccg cgcatcttca ccttgtagat
     4261 cagcgtgccg tcctgcaggg aggagtcctg ggtcacggtc accagaccgc cgtcctcgaa
     4321 gttcatcacg cgctcccact tgaagccctc ggggaaggac agcttcttgt aatcggggat
     4381 gtcggcgggg tgcttcacgt acgccttgga gccgtacatg aactgggggg acaggatgtc
     4441 ccaggcgaag ggcagggggc cgcccttggt caccttcagc ttggcggtct gggtgccctc
     4501 gtaggggcgg ccctcgccct cgccctcgat ctcgaactcg tggccgttca tggagccctc
     4561 catgcgcacc ttgaagcgca tgaactcttt gatgacggcc atgttgttgt cctcggagga
     4621 ggcggtgccg gagctgccgc taccggtgct gccggtgcca tgccccagga acaggtggtg
     4681 gcggccctcg gagcgctcgt actgttccac gatggtgtag tcctcgttgt gggaggtgat
     4741 gtccagcttg gtgtccacgt agtagtagcc gggcagttgc acgggcttct tggccatgta
     4801 gatggtcttg aactccacca ggtagtggcc gccgtccttc agcttcaggg cctggtggat
     4861 ctcgcccttc agcacgccgt cgcgggggta caggcgctcg gtggaggcct cccagcccat
     4921 ggtcttcttc tgcattacgg ggccgtcggg ggggaagttg gtgccgcgca tcttcacctt
     4981 gtagatcagc gtgccgtcct gcagggagga gtcctgggtc acggtcacca gaccgccgtc
     5041 ctcgaagttc atcacgcgct cccacttgaa gccctcgggg aaggacagct tcttgtaatc
     5101 ggggatgtcg gcggggtgct tcacgtacgc cttggagccg tacatgaact ggggggacag
     5161 gatgtcccag gcgaagggca gggggccgcc cttggtcacc ttcagcttgg cggtctgggt
     5221 gccctcgtag gggcggccct cgccctcgcc ctcgatctcg aactcgtggc cgttcatgga
     5281 gccctccatg cgcaccttga agcgcatgaa ctctttgatg acctcctcgc ccttgctcac
     5341 catggttccg gaagggccgg gattctcctc cacgtcaccg catgttagaa gacttcctct
     5401 gccctcagcg gccgctaaca cggcgatctt gccgcccttc ttggccttaa tgagaatctc
     5461 gcggatcttg cgggcgtcca acttgccggt cagtccttta ggcacctcgt ccacgaacac
     5521 aacaccaccg cgcagcttct tggcggttgt aacctggctg gccacatagt ccacgatctc
     5581 cttctcggtc atggttttac cgtgttccag cacgacgact gcggcgggca gctcgccggc
     5641 atcgtcgtcg ggcaggccgg cgaccccggc gtcgaagatg ttggggtgtt gcagcaggat
     5701 gctctccagt tcggctgggg ctacctggta gcccttgtat ttgatcaggc tcttcagccg
     5761 gtccacgatg aagaagtgct cgtcctcgtc ccagtaggcg atgtcgccgc tgtgcagcca
     5821 gccgtccttg tcgatgagag cgtttgtagc ctcggggttg ttaacgtagc cgctcatgat
     5881 catggggcca cggacgcaca gctcgccgcg ctggttcaca cccagtgtct taccggtgtc
     5941 caagtccacc accttagcct cgaagaaggg caccaccttg cctactgcgc caggcttgtc
     6001 gtccccttcg ggggtgatca gaatggcgct ggttgtttct gtcaggccgt agccctggcg
     6061 gatgcctggt aggtggaagc gtttggccac ggcctcacct acctccttgc tgagcggcgc
     6121 cccgccgctg gcgatctcgt gcaagttgct taggtcgtac ttgtcgatga gagtgctctt
     6181 agcgaagaag ctaaatagtg tgggcaccag cagggcagat tgaatcttat agtcttgcaa
     6241 gctgcgcaag aatagctcct cctcgaagcg gtacatgagc acgacccgaa agccgcagat
     6301 caagtagccc agcgtggtga acatgccgaa gccgtggtga aatggcacca cgctgaggat
     6361 agcggtgtcg gggatgatct ggttgccgaa gatggggtcg cgggcatgac tgaatcggac
     6421 acaagcggtg cggtgcggta gggctacgcc cttgggcaat ccggtactgc cactactgtt
     6481 catgatcagg gcgatggttt tgtcccggtc gaagctctcg ggcacgaagt cgtactcgtt
     6541 gaagccgggt ggcaaatggg aagtcacgaa ggtgtacatg ctttggaagc cctggtagtc
     6601 ggtcttgcta tccatgatga tgatcttttg tatgatcggt agcttctttt gcacgttgag
     6661 gatcttttgc agccctttct tgctcacgaa tacgacggtg ggctggctga tgcccatgct
     6721 gttcagcagc tcgcgctcgt tgtagatgtc gttagctggg gccacagcca caccgatgaa
     6781 cagggcaccc aacacgggca tgaagaactg caagctattc tcgctgcaca ccacgatccg
     6841 atggtttgta ttcagcccat agcgcttcat agcttctgcc agccgaacgc tcatctcgaa
     6901 gtactcggcg taggtaatgt ccacctcgat atgtgcgtcg gtaaaggcga tggtgccggg
     6961 caccagggcg tagcgcttca tggctttgtg cagctgctcg ccggcggtcc cgtcttcgag
     7021 tgggtagaat ggcgctgggc ccttcttaat gtttttggca tcttccatga attcgctagc
     7081 tctagagtag gcgccggtca cagcttggat ctgtaacggc gcagaacaga aaacgaaaca
     7141 aagacgtaga gttgagcaag cagggtcagg caaagcgtgg agagccggct gagtctaggt
     7201 aggctccaag ggagcgccgg acaaaggccc ggtctcgacc tgagctttaa acttacctag
     7261 acggcggacg cagttcagga ggcaccacag gcgggaggcg gcagaacgcg actcaaccgg
     7321 cgtggatggc ggcctcaggt agggcggcgg gcgcgtgaag gagagatgcg agcccctcga
     7381 agcttcagct gtgttctggc ggcaaacccg ttgcgaaaaa gaacgttcac ggcgactact
     7441 gcacttatat acggttctcc cccaccctcg ggaaaaaggc ggagccagta cacgacatca
     7501 ctttcccagt ttaccccgcg ccaccttctc taggcacccg ttcaattgcc gacccctccc
     7561 cccaacttct cggggactgt gggcgatgtg cgctctgccc actgacgggc accggagcga
     7621 tcgcagatcc ttactagtat cgataaaatt ttgaattttt gtaatttgtt tttgtaattc
     7681 tttagtttgt atgtctgttg ctattatgtc tactattctt tcccctgcac tgtacccccc
     7741 aatcccccct tttcttttaa aagttaaccg ataccgtcga gatccgttca ctaatcgaat
     7801 ggatctgtct ctgtctctct ctccaccttc ttcttctatt ccttcgggcc tgtcgggtcc
     7861 cctcggggtt gggaggtggg tctgaaacga taatggtgaa tatccctgcc taactctatt
     7921 cactatagaa agtacagcaa aaactattct taaacctacc aagcctccta ctatcattat
     7981 gaataatttt atataccaca gccaatttgt tatgttaaac caattccaca aacttgccca
     8041 tttatctaat tccaataatt cttgttcatt cttttcttgc tggttttgcg attcttcaat
     8101 taaggagtgt attaagcttg tgtaattgtt aatttctctg tcccactcca tccaggtcgt
     8161 gtgattccaa tctgttccag agatttatta ctccaactag cattccaagg cacagcagtg
     8221 gtgcaaatga gttttccaga gcaaccccaa atccccagga gctgttgatc ctttaggtat
     8281 ctttccacag ccaggattct tgcctggagc tgcttgatgc cccagactgt gagttgcaac
     8341 agatgctgtt gcgcctcaat agccctcagc aaattgttct gctgctgcac tataccagac
     8401 aataattgtc tggcctgtac cgtcagcgtc attgaggctg cgcccatagt gcttcctgct
     8461 gctcccaaga acccaaggaa caaagctcct attcccactg ctcttttttc tctctgcacc
     8521 actcttctct ttgccttggt gggtgctact cctaatggtt caatttttac tactttatat
     8581 ttatataatt cacttctcca attgtccctc atatctcctc ctccaggtct gaagatcagt
     8641 ggccgcttgc tgtgcggtgg tcttactttt gttttgctct tcctctatct tgtctaaagc
     8701 ttccttggtg tcttttatct ctatcctttg atgcacacaa tagagggttg ctactgtatt
     8761 atataatgat ctaagttctt ctgatcctgt ctgaagggat ggttgtagct gtcccagtat
     8821 ttgtctacag ccttctgatg tttctaacag gccaggatta actgcgaatc gttctagctc
     8881 cctgcttgcc catactatat gttttaattt atattttttc tttccccctg gccttaaccg
     8941 aattttttcc catcgcgatc taattctccc ccgcttaata ctgacgctct cgcacccatc
     9001 tctctccttc tagcctccgc tagtcaaaat ttttggcgta ctcaccagtc gccgcccctc
     9061 gcctcttgcc gtgcgcgctt cagcaagccg agtcctgcgt cgagagagct ctggtttccc
     9121 tttcgctttc aggtccctgt tcgggcgcca ctgctagaga ttttccacac tgactaaaag
     9181 ggtctgaggg atctctagtt accagagtca cacaacagac gggcacacac tacttgaagc
     9241 actcaaggca agctttattg aggcttaagc agtgggttcc ctagttagcc agagagctcc
     9301 caggctcaga tctggtctaa ccagagagac ccgtttattg tatcgagcta ggcacttaaa
     9361 tacaatatct ctgcaatgcg gcaattcagt ggttcgtcca atccatgtca gacccgtctg
     9421 ttgccttcct aataaggcac gatcgtacca ccttacttcc accaatcggc atgcacggtg
     9481 ctttttctct ccttgtaagg catgttgcta actcatcgtt accatgttgc aagactacaa
     9541 gagtattgca taagactaca cgcgt