pCIG哺乳表达质粒

基本信息

启动子:	CAG promoter
复制子:	Ori，SV40 ori
终止子:	β-globin poly(A) signal
质粒分类:	哺乳细胞质粒；哺乳表达质粒；pCAG系列质粒
质粒大小:	6200bp
原核抗性:	氨苄青霉素Amp
筛选标记:	EGFP
克隆菌株:	DH5α等大肠杆菌
培养条件:	37℃，有氧，LB
表达宿主:	293T等哺乳细胞

质粒简介

         pCIG质粒是一个哺乳细胞过表达载体，CAG启动子驱动目的基因的表达。

质粒图谱

质粒序列

LOCUS       Exported                6200 bp ds-DNA    circular SYN 18-八月-2017
EYWORDS    pCIG
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6200)
FEATURES             Location/Qualifiers
     source          1..6200
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      complement(42..630)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(801..1661)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(1662..1766)
                     /gene="bla"
                     /note="AmpR promoter"
     enhancer        1797..2176
                     /note="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2178..2455
                     /note="chicken beta-actin promoter"
     intron          2456..3468
                     /note="chimeric intron"
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     misc_feature    3594..4172
                     /note="IRES2"
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             4173..4892
                     /codon_start=1
                     /product="enhanced GFP"
                     /note="EGFP"
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     polyA_site      5049..5104
                     /note="beta-globin poly(A) signal"
                     /note="rabbit beta-globin polyadenylation signal"
     promoter        5668..5863
                     /note="SV40 promoter"
                     /note="SV40 early promoter"
     rep_origin      5714..5849
                     /note="SV40 ori"
                     /note="SV40 origin of replication"
     polyA_signal    5869..6003
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
       61 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
      121 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
      181 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
      241 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
      301 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
      361 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
      421 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
      481 agnacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
      541 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
      601 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
      661 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
      721 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
      781 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
      841 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
      901 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
      961 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     1021 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     1081 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     1141 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     1201 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     1261 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     1321 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     1381 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     1441 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     1501 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     1561 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     1621 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     1681 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     1741 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgggtcgaca
     1801 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
     1861 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
     1921 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
     1981 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
     2041 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     2101 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
     2161 catcgctatt accatggtcg aggtgagccc cacgttctgc ttcactctcc ccatctcccc
     2221 cccctcccca cccccaattt tgtatttatt tattttttaa ttattttgtg cagcgatggg
     2281 ggcggggggg gggggggggc gcgcgccagg cggggcgggg cggggcgagg ggcggggcgg
     2341 ggcgaggcgg agaggtgcgg cggcagccaa tcagagcggc gcgctccgaa agtttccttt
     2401 tatggcgagg cggcggcggc ggcggcccta taaaaagcga agcgcgcggc gggcgggagt
     2461 cgctgcgttg ccttcgcccc gtgccccgct ccgcgccgcc tcgcgccgcc cgccccggct
     2521 ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc ctccgggctg
     2581 taattagcgc ttggtttaat gacggctcgt ttcttttctg tggctgcgtg aaagccttaa
     2641 agggctccgg gagggccctt tgtgcggggg ggagcggctc ggggggtgcg tgcgtgtgtg
     2701 tgtgcgtggg gagcgccgcg tgcggcccgc gctgcccggc ggctgtgagc gctgcgggcg
     2761 cggcgcgggg ctttgtgcgc tccgcgtgtg cgcgagggga gcgcggccgg gggcggtgcc
     2821 ccgcggtgcg ggggggctgc gaggggaaca aaggctgcgt gcggggtgtg tgcgtggggg
     2881 ggtgagcagg gggtgtgggc gcggcggtcg ggctgtaacc cccccctgca cccccctccc
     2941 cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg tggcgcgggg
     3001 ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc ggggccgcct
     3061 cgggccgggg agggctcggg ggaggggcgc ggcggccccc ggagcgccgg cggctgtcga
     3121 ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc gcagggactt
     3181 cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac cccctctagc
     3241 gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga gggccttcgt
     3301 gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc gcggggggac
     3361 ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt gaccggcggc
     3421 tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct cctgggcaac
     3481 gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tgctcgaggt cgatcgacgg
     3541 tatcgataag cttgatatcg aattcctgca gcccggccgc tacgtaaatt ccgccccccc
     3601 cccccctaac gttactggcc gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat
     3661 gttattttcc accatattgc cgtcttttgg caatgtgagg gcccggaaac ctggccctgt
     3721 cttcttgacg agcattccta ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt
     3781 gaatgtcgtg aaggaagcag ttcctctgga agcttcttga agacaaacaa cgtctgtagc
     3841 gaccctttgc aggcagcgga accccccacc tggcgacagg tgcctctgcg gccaaaagcc
     3901 acgtgtataa gatacacctg caaaggcggc acaaccccag tgccacgttg tgagttggat
     3961 agttgtggaa agagtcaaat ggctctcctc aagcgtattc aacaaggggc tgaaggatgc
     4021 ccagaaggta ccccattgta tgggatctga tctggggcct cggtacacat gctttacatg
     4081 tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac cacggggacg tggttttcct
     4141 ttgaaaaaca cgatgataat atggccacaa ccatggtgag caagggcgag gagctgttca
     4201 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
     4261 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca
     4321 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc
     4381 agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
     4441 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
     4501 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
     4561 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca
     4621 acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc
     4681 acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     4741 gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca
     4801 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
     4861 tcactctcgg catggacgag ctgtacaagt aaagcggccg caattcactc ctcaggtgca
     4921 ggctgcctat cagaaggtgg tggctggtgt ggccaatgcc ctggctcaca aataccactg
     4981 agatcttttt ccctctgcca aaaattatgg ggacatcatg aagccccttg agcatctgac
     5041 ttctggctaa taaaggaaat ttattttcat tgcaatagtg tgttggaatt ttttgtgtct
     5101 atcactcgga aggacatatg ggagggcaaa tcatttaaaa catcagaatg agtatttggt
     5161 ttagagtttg gcaacatatg cccatatgct ggctgccatg aacaaaggtt ggctataaag
     5221 aggtcatcag tatatgaaac agccccctgc tgtccattcc ttattccata gaaaagcctt
     5281 gacttgaggt tagatttttt ttatattttg ttttgtgtta tttttttctt taacatccct
     5341 aaaattttcc ttacatgttt tactagccag atttttcctc ctctcctgac tactcccagt
     5401 catagctgtc cctcttctct tatggagatc cctcgacctg caccgtcgac cagctggtcg
     5461 acggtgcacc gtcgaccagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     5521 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     5581 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     5641 cgggaaacct gtcgtgccag cggatccgca tctcaattag tcagcaacca tagtcccgcc
     5701 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     5761 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     5821 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctaa cttgtttatt
     5881 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     5941 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg
     6001 atccgctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     6061 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     6121 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     6181 acatgtgagc aaaaggccag