LOCUS       V042760                 7872 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V042760
VERSION     V042760
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     promoter        123..1301
                     /note="strong constitutive promoter for human elongation
                     factor EF-1α"
                     /label="EF-1α promoter"
     CDS             1352..3829
                     /label="HIF1A(NM_001530)"
                     /note="HIF1A(NM_001530)"
                     /gene="HIF1A"
     CDS             3872..4591
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /transl_table=1
                     /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
                     TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG
                     IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV
                     LLEFVTAAGITLGMDELYK*"
                     /label="EGFP"
     polyA_signal    4713..4834
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      complement(4841..5296)
                     /direction=LEFT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5323..5427
                     /gene="bla"
                     /label="AmpR promoter"
     promoter        5429..5786
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5821..6615
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin®)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /transl_table=1
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    6847..6894
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      7223..7811
                     /direction=RIGHT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
ORIGIN
        1 cgcgttgaca ttgattattg actaggcttt tgcaaaaagc tttgcaaaga tggataaagt
       61 tttaaacaga gaggaatctt tgcagctaat ggaccttcta ggtcttgaaa ggagtgggaa
      121 ttggctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg
      181 ggggaggggt cggcaattga accggtgcct agagaaggtg gcgcggggta aactgggaaa
      241 gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg
      301 cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca caggtaagtg
      361 ccgtgtgtgg ttcccgcggg cctggcctct ttacgggtta tggcccttgc gtgccttgaa
      421 ttacttccac ctggctgcag tacgtgattc ttgatcccga gcttcgggtt ggaagtgggt
      481 gggagagttc gaggccttgc gcttaaggag ccccttcgcc tcgtgcttga gttgaggcct
      541 ggcctgggcg ctggggccgc cgcgtgcgaa tctggtggca ccttcgcgcc tgtctcgctg
      601 ctttcgataa gtctctagcc atttaaaatt tttgatgacc tgctgcgacg ctttttttct
      661 ggcaagatag tcttgtaaat gcgggccaag atctgcacac tggtatttcg gtttttgggg
      721 ccgcgggcgg cgacggggcc cgtgcgtccc agcgcacatg ttcggcgagg cggggcctgc
      781 gagcgcggcc accgagaatc ggacgggggt agtctcaagc tggccggcct gctctggtgc
      841 ctggcctcgc gccgccgtgt atcgccccgc cctgggcggc aaggctggcc cggtcggcac
      901 cagttgcgtg agcggaaaga tggccgcttc ccggccctgc tgcagggagc tcaaaatgga
      961 ggacgcggcg ctcgggagag cgggcgggtg agtcacccac acaaaggaaa agggcctttc
     1021 cgtcctcagc cgtcgcttca tgtgactcca cggagtaccg ggcgccgtcc aggcacctcg
     1081 attagttctc gagcttttgg agtacgtcgt ctttaggttg gggggagggg ttttatgcga
     1141 tggagtttcc ccacactgag tgggtggaga ctgaagttag gccagcttgg cacttgatgt
     1201 aattctcctt ggaatttgcc ctttttgagt ttggatcttg gttcattctc aagcctcaga
     1261 cagtggttca aagttttttt cttccatttc aggtgtcgtg aggaattagc ttggtacgct
     1321 agcgtttaaa cttaagcttg gtaccgccac catggagggc gccggcggcg cgaacgacaa
     1381 gaaaaagata agttctgaac gtcgaaaaga aaagtctcga gatgcagcca gatctcggcg
     1441 aagtaaagaa tctgaagttt tttatgagct tgctcatcag ttgccacttc cacataatgt
     1501 gagttcgcat cttgataagg cctctgtgat gaggcttacc atcagctatt tgcgtgtgag
     1561 gaaacttctg gatgctggtg atttggatat tgaagatgac atgaaagcac agatgaattg
     1621 cttttatttg aaagccttgg atggttttgt tatggttctc acagatgatg gtgacatgat
     1681 ttacatttct gataatgtga acaaatacat gggattaact cagtttgaac taactggaca
     1741 cagtgtgttt gattttactc atccatgtga ccatgaggaa atgagagaaa tgcttacaca
     1801 cagaaatggc cttgtgaaaa agggtaaaga acaaaacaca cagcgaagct tttttctcag
     1861 aatgaagtgt accctaacta gccgaggaag aactatgaac ataaagtctg caacatggaa
     1921 ggtattgcac tgcacaggcc acattcacgt atatgatacc aacagtaacc aacctcagtg
     1981 tgggtataag aaaccaccta tgacctgctt ggtgctgatt tgtgaaccca ttcctcaccc
     2041 atcaaatatt gaaattcctt tagatagcaa gactttcctc agtcgacaca gcctggatat
     2101 gaaattttct tattgtgatg aaagaattac cgaattgatg ggatatgagc cagaagaact
     2161 tttaggccgc tcaatttatg aatattatca tgctttggac tctgatcatc tgaccaaaac
     2221 tcatcatgat atgtttacta aaggacaagt caccacagga cagtacagga tgcttgccaa
     2281 aagaggtgga tatgtctggg ttgaaactca agcaactgtc atatataaca ccaagaattc
     2341 tcaaccacag tgcattgtat gtgtgaatta cgttgtgagt ggtattattc agcacgactt
     2401 gattttctcc cttcaacaaa cagaatgtgt ccttaaaccg gttgaatctt cagatatgaa
     2461 aatgactcag ctattcacca aagttgaatc agaagataca agtagcctct ttgacaaact
     2521 taagaaggaa cctgatgctt taactttgct ggccccagcc gctggagaca caatcatatc
     2581 tttagatttt ggcagcaacg acacagaaac tgatgaccag caacttgagg aagtaccatt
     2641 atataatgat gtaatgctcc cctcacccaa cgaaaaatta cagaatataa atttggcaat
     2701 gtctccatta cccaccgctg aaacgccaaa gccacttcga agtagtgctg accctgcact
     2761 caatcaagaa gttgcattaa aattagaacc aaatccagag tcactggaac tttcttttac
     2821 catgccccag attcaggatc agacacctag tccttccgat ggaagcacta gacaaagttc
     2881 acctgagcct aatagtccca gtgaatattg tttttatgtg gatagtgata tggtcaatga
     2941 attcaagttg gaattggtag aaaaactttt tgctgaagac acagaagcaa agaacccatt
     3001 ttctactcag gacacagatt tagacttgga gatgttagct ccctatatcc caatggatga
     3061 tgacttccag ttacgttcct tcgatcagtt gtcaccatta gaaagcagtt ccgcaagccc
     3121 tgaaagcgca agtcctcaaa gcacagttac agtattccag cagactcaaa tacaagaacc
     3181 tactgctaat gccaccacta ccactgccac cactgatgaa ttaaaaacag tgacaaaaga
     3241 ccgtatggaa gacattaaaa tattgattgc atctccatct cctacccaca tacataaaga
     3301 aactactagt gccacatcat caccatatag agatactcaa agtcggacag cctcaccaaa
     3361 cagagcagga aaaggagtca tagaacagac agaaaaatct catccaagaa gccctaacgt
     3421 gttatctgtc gctttgagtc aaagaactac agttcctgag gaagaactaa atccaaagat
     3481 actagctttg cagaatgctc agagaaagcg aaaaatggaa catgatggtt cactttttca
     3541 agcagtagga attggaacat tattacagca gccagacgat catgcagcta ctacatcact
     3601 ttcttggaaa cgtgtaaaag gatgcaaatc tagtgaacag aatggaatgg agcaaaagac
     3661 aattatttta ataccctctg atttagcatg tagactgctg gggcaatcaa tggatgaaag
     3721 tggattacca cagctgacca gttatgattg tgaagttaat gctcctatac aaggcagcag
     3781 aaacctactg cagggtgaag aattactcag agctttggat caagttaacg gtggaggcgg
     3841 atcaggaggc ggtggatctg gatccgccac catggtgagc aagggcgagg agctgttcac
     3901 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
     3961 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac
     4021 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca
     4081 gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
     4141 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
     4201 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     4261 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
     4321 cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca
     4381 caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
     4441 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa
     4501 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
     4561 cactctcggc atggacgagc tgtacaagta agcggccgcg actctagatc ataatcagcc
     4621 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
     4681 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
     4741 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     4801 gttgtggttt gtccaaactc atcaatgtat cttaaggcgt aaattgtaag cgttaatatt
     4861 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     4921 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     4981 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     5041 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     5101 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     5161 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     5221 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     5281 ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     5341 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     5401 cttcaataat attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt
     5461 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     5521 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     5581 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     5641 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     5701 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     5761 tttttggagg cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc
     5821 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     5881 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     5941 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     6001 caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     6061 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     6121 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     6181 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     6241 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     6301 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac
     6361 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     6421 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     6481 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     6541 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     6601 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     6661 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     6721 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     6781 cccaccctag ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg
     6841 ctatgacggc aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac
     6901 gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc
     6961 aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag
     7021 ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac
     7081 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     7141 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     7201 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     7261 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     7321 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     7381 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     7441 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     7501 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     7561 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     7621 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     7681 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     7741 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     7801 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     7861 ttgctcacat gt
//
