pCDH-CMV-MCS-EF1-Puro-FLAG-HIF

基本信息

别名:	NM_001430.4；EPAS1；bHLHe73; ECYT4;
原核抗性:	Amp
筛选标记:	Puro
克隆菌株:	Stbl3
培养条件:	LB/37℃

质粒简介

pCDH-CMV-MCS-EF1-Puro-Flag-HIF2α人源基因质粒

质粒图谱

质粒序列

LOCUS       Exported               10010 bp ds-DNA     circular SYN 06-JUL-2018
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pCDH-CMV-MCS-EF1-Puro-Flag-HIF2-alpha
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10010)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..10010
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     LTR             234..414
                     /note="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    458..583
                     /note="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1076..1309
                     /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    1804..1920
                     /note="cPPT/CTS"
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2010..2213
                     /note="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     regulatory      2290..2299
                     /regulatory_class="other"
                     /note="Kozak sequence"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             2299..2322
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /note="FLAG"
                     /translation="DYKDDDDK"
     misc_feature    2323..4932
                     /note="HIF2-alpha"
     promoter        4980..5191
                     /note="EF-1-alpha core promoter"
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             5204..5472
                     /note="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             5499..6098
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /note="PuroR"
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     misc_feature    6105..6693
                     /note="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     LTR             6767..7000
                     /note="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    7072..7193
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      7212..7347
                     /note="SV40 ori"
                     /note="SV40 origin of replication"
     rep_origin      complement(7787..8375)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(8546..9406)
                     /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(9407..9511)
                     /gene="bla"
                     /note="AmpR promoter"
ORIGIN
        1 acgcgtgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca
       61 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta
      121 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga
      181 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacaata aacgggtctc
      241 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
      301 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
      361 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg
      421 cccgaacagg gacctgaaag cgaaagggaa accagagctc tctcgacgca ggactcggct
      481 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt
      541 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag
      601 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt
      661 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt
      721 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg
      781 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag
      841 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag
      901 taagaccacc gcacagcaag cggccactga tcttcagacc tggaggagga gatatgaggg
      961 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag
     1021 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag
     1081 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcc tcaatgacgc
     1141 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga
     1201 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc
     1261 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg
     1321 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata
     1381 aatctctgga acagattgga atcacacgac ctggatggag tgggacagag aaattaacaa
     1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
     1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
     1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
     1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
     1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcggtt
     1801 aacttttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat
     1861 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaat tcaaaatttt
     1921 atcgatacta gtattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     1981 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
     2041 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
     2101 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
     2161 tgacgcaaat gggcggtagg cgtgtacggt gggaggttta tataagcaga gctcgtttag
     2221 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagattct
     2281 agagctagcg ccaccatgga ttacaaggat gacgacgata agacagctga caaggagaag
     2341 aaaaggagta gctcggagag gaggaaggag aagtcccggg atgctgcgcg gtgccggcgg
     2401 agcaaggaga cggaggtgtt ctatgagctg gcccatgagc tgcctctgcc ccacagtgtg
     2461 agctcccatc tggacaaggc ctccatcatg cgactggcaa tcagcttcct gcgaacacac
     2521 aagctcctct cctcagtttg ctctgaaaac gagtccgaag ccgaagctga ccagcagatg
     2581 gacaacttgt acctgaaagc cttggagggt ttcattgccg tggtgaccca agatggcgac
     2641 atgatctttc tgtcagaaaa catcagcaag ttcatgggac ttacacaggt ggagctaaca
     2701 ggacatagta tctttgactt cactcatccc tgcgaccatg aggagattcg tgagaacctg
     2761 agtctcaaaa atggctctgg ttttgggaaa aaaagcaaag acatgtccac agagcgggac
     2821 ttcttcatga ggatgaagtg cacggtcacc aacagaggcc gtactgtcaa cctcaagtca
     2881 gccacctgga aggtcttgca ctgcacgggc caggtgaaag tctacaacaa ctgccctcct
     2941 cacaatagtc tgtgtggcta caaggagccc ctgctgtcct gcctcatcat catgtgtgaa
     3001 ccaatccagc acccatccca catggacatc cccctggata gcaagacctt cctgagccgc
     3061 cacagcatgg acatgaagtt cacctactgt gatgacagaa tcacagaact gattggttac
     3121 caccctgagg agctgcttgg ccgctcagcc tatgaattct accatgcgct agactccgag
     3181 aacatgacca agagtcacca gaacttgtgc accaagggtc aggtagtaag tggccagtac
     3241 cggatgctcg caaagcatgg gggctacgtg tggctggaga cccaggggac ggtcatctac
     3301 aaccctcgca acctgcagcc ccagtgcatc atgtgtgtca actacgtcct gagtgagatt
     3361 gagaagaatg acgtggtgtt ctccatggac cagactgaat ccctgttcaa gccccacctg
     3421 atggccatga acagcatctt tgatagcagt ggcaaggggg ctgtgtctga gaagagtaac
     3481 ttcctattca ccaagctaaa ggaggagccc gaggagctgg cccagctggc tcccacccca
     3541 ggagacgcca tcatctctct ggatttcggg aatcagaact tcgaggagtc ctcagcctat
     3601 ggcaaggcca tcctgccccc gagccagcca tgggccacgg agttgaggag ccacagcacc
     3661 cagagcgagg ctgggagcct gcctgccttc accgtgcccc aggcagctgc cccgggcagc
     3721 accaccccca gtgccaccag cagcagcagc agctgctcca cgcccaatag ccctgaagac
     3781 tattacacat ctttggataa cgacctgaag attgaagtga ttgagaagct cttcgccatg
     3841 gacacagagg ccaaggacca atgcagtacc cagacggatt tcaatgagct ggacttggag
     3901 acactggcac cctatatccc catggacggg gaagacttcc agctaagccc catctgcccc
     3961 gaggagcggc tcttggcgga gaacccacag tccacccccc agcactgctt cagtgccatg
     4021 acaaacatct tccagccact ggcccctgta gccccgcaca gtcccttcct cctggacaag
     4081 tttcagcagc agctggagag caagaagaca gagcccgagc accggcccat gtcctccatc
     4141 ttctttgatg ccggaagcaa agcatccctg ccaccgtgct gtggccaggc cagcacccct
     4201 ctctcttcca tggggggcag atccaatacc cagtggcccc cagatccacc attacatttt
     4261 gggcccacaa agtgggccgt cggggatcag cgcacagagt tcttgggagc agcgccgttg
     4321 gggccccctg tctctccacc ccatgtctcc accttcaaga caaggtctgc aaagggtttt
     4381 ggggctcgag gcccagacgt gctgagtccg gccatggtag ccctctccaa caagctgaag
     4441 ctgaagcgac agctggagta tgaagagcaa gccttccagg acctgagcgg gggggaccca
     4501 cctggtggca gcacctcaca tttgatgtgg aaacggatga agaacctcag gggtgggagc
     4561 tgccctttga tgccggacaa gccactgagc gcaaatgtac ccaatgataa gttcacccaa
     4621 aaccccatga ggggcctggg ccatcccctg agacatctgc cgctgccaca gcctccatct
     4681 gccatcagtc ccggggagaa cagcaagagc aggttccccc cacagtgcta cgccacccag
     4741 taccaggact acagcctgtc gtcagcccac aaggtgtcag gcatggcaag ccggctgctc
     4801 gggccctcat ttgagtccta cctgctgccc gaactgacca gatatgactg tgaggtgaac
     4861 gtgcccgtgc tgggaagctc cacgctcctg caaggagggg acctcctcag agccctggac
     4921 caggccacct gaggatccgc ggccgcgaag gatctgcgat cgctccggtg cccgtcagtg
     4981 ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg gcaattgaac
     5041 gggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg
     5101 cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg tgaacgttct
     5161 ttttcgcaac gggtttgccg ccagaacaca gctgaagctt cgaggggctc gcatctctcc
     5221 ttcacgcgcc cgccgcccta cctgaggccg ccatccacgc cggttgagtc gcgttctgcc
     5281 gcctcccgcc tgtggtgcct cctgaactgc gtccgccgtc taggtaagtt taaagctcag
     5341 gtcgagaccg ggcctttgtc cggcgctccc ttggagccta cctagactca gccggctctc
     5401 cacgctttgc ctgaccctgc ttgctcaact ctacgtcttt gtttcgtttt ctgttctgcg
     5461 ccgttacaga tccaagctgt gaccggcgcc tacgctagat gaccgagtac aagcccacgg
     5521 tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc gccgcgttcg
     5581 ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag cgggtcaccg
     5641 agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg
     5701 acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt
     5761 tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac
     5821 agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg
     5881 tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag
     5941 tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc
     6001 tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac
     6061 cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgagt cgacaatcaa cctctggatt
     6121 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     6181 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
     6241 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc
     6301 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca
     6361 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac
     6421 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
     6481 ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct
     6541 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc
     6601 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga
     6661 cgagtcggat ctccctttgg gccgcctccc cgcctggtac ctttaagacc aatgacttac
     6721 aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt
     6781 cactcccaac gaaaataaga tctgcttttt gcttgtactg ggtctctctg gttagaccag
     6841 atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc
     6901 ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga
     6961 tccctcagac ccttttagtc agtgtggaaa atctctagca gtagtagttc atgtcatctt
     7021 attattcagt atttataact tgcaaagaaa tgaatatcag agagtgagag gaacttgttt
     7081 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     7141 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     7201 tggctctagc tatcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     7261 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     7321 agtagtgagg aggctttttt ggaggcctag acttttgcag agacggccca aattcgtaat
     7381 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     7441 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     7501 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     7561 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     7621 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     7681 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     7741 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     7801 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     7861 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     7921 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     7981 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     8041 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     8101 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     8161 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     8221 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     8281 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     8341 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     8401 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     8461 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     8521 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     8581 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     8641 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     8701 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     8761 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     8821 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     8881 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     8941 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     9001 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     9061 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     9121 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     9181 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     9241 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     9301 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     9361 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     9421 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     9481 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     9541 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
     9601 ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga
     9661 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
     9721 cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga
     9781 gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca
     9841 ggcgccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt
     9901 cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc
     9961 cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgccaagctg