LOCUS       V032480                 7670 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032480
VERSION     V032480
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             908..3385
                     /label="HIF1A(NM_001530)"
                     /note="HIF1A(NM_001530)"
                     /gene="HIF1A"
     polyA_signal    3432..3656
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3702..4130
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4144..4473
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4540..5139
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /note="confers resistance to puromycin"
                     /product="puromycin N-acetyltransferase"
                     /transl_table=1
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*"
                     /label="PuroR"
     polyA_signal    5313..5434
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5483..5499)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5507..5523
                     /bound_moiety="lac repressor encoded by lacI"
                     /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-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        complement(5531..5561)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5576..5597
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5885..6470)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6641..7501)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     promoter        complement(7502..7606)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg agctcggatc
      901 cgccaccatg gagggcgccg gcggcgcgaa cgacaagaaa aagataagtt ctgaacgtcg
      961 aaaagaaaag tctcgagatg cagccagatc tcggcgaagt aaagaatctg aagtttttta
     1021 tgagcttgct catcagttgc cacttccaca taatgtgagt tcgcatcttg ataaggcctc
     1081 tgtgatgagg cttaccatca gctatttgcg tgtgaggaaa cttctggatg ctggtgattt
     1141 ggatattgaa gatgacatga aagcacagat gaattgcttt tatttgaaag ccttggatgg
     1201 ttttgttatg gttctcacag atgatggtga catgatttac atttctgata atgtgaacaa
     1261 atacatggga ttaactcagt ttgaactaac tggacacagt gtgtttgatt ttactcatcc
     1321 atgtgaccat gaggaaatga gagaaatgct tacacacaga aatggccttg tgaaaaaggg
     1381 taaagaacaa aacacacagc gaagcttttt tctcagaatg aagtgtaccc taactagccg
     1441 aggaagaact atgaacataa agtctgcaac atggaaggta ttgcactgca caggccacat
     1501 tcacgtatat gataccaaca gtaaccaacc tcagtgtggg tataagaaac cacctatgac
     1561 ctgcttggtg ctgatttgtg aacccattcc tcacccatca aatattgaaa ttcctttaga
     1621 tagcaagact ttcctcagtc gacacagcct ggatatgaaa ttttcttatt gtgatgaaag
     1681 aattaccgaa ttgatgggat atgagccaga agaactttta ggccgctcaa tttatgaata
     1741 ttatcatgct ttggactctg atcatctgac caaaactcat catgatatgt ttactaaagg
     1801 acaagtcacc acaggacagt acaggatgct tgccaaaaga ggtggatatg tctgggttga
     1861 aactcaagca actgtcatat ataacaccaa gaattctcaa ccacagtgca ttgtatgtgt
     1921 gaattacgtt gtgagtggta ttattcagca cgacttgatt ttctcccttc aacaaacaga
     1981 atgtgtcctt aaaccggttg aatcttcaga tatgaaaatg actcagctat tcaccaaagt
     2041 tgaatcagaa gatacaagta gcctctttga caaacttaag aaggaacctg atgctttaac
     2101 tttgctggcc ccagccgctg gagacacaat catatcttta gattttggca gcaacgacac
     2161 agaaactgat gaccagcaac ttgaggaagt accattatat aatgatgtaa tgctcccctc
     2221 acccaacgaa aaattacaga atataaattt ggcaatgtct ccattaccca ccgctgaaac
     2281 gccaaagcca cttcgaagta gtgctgaccc tgcactcaat caagaagttg cattaaaatt
     2341 agaaccaaat ccagagtcac tggaactttc ttttaccatg ccccagattc aggatcagac
     2401 acctagtcct tccgatggaa gcactagaca aagttcacct gagcctaata gtcccagtga
     2461 atattgtttt tatgtggata gtgatatggt caatgaattc aagttggaat tggtagaaaa
     2521 actttttgct gaagacacag aagcaaagaa cccattttct actcaggaca cagatttaga
     2581 cttggagatg ttagctccct atatcccaat ggatgatgac ttccagttac gttccttcga
     2641 tcagttgtca ccattagaaa gcagttccgc aagccctgaa agcgcaagtc ctcaaagcac
     2701 agttacagta ttccagcaga ctcaaataca agaacctact gctaatgcca ccactaccac
     2761 tgccaccact gatgaattaa aaacagtgac aaaagaccgt atggaagaca ttaaaatatt
     2821 gattgcatct ccatctccta cccacataca taaagaaact actagtgcca catcatcacc
     2881 atatagagat actcaaagtc ggacagcctc accaaacaga gcaggaaaag gagtcataga
     2941 acagacagaa aaatctcatc caagaagccc taacgtgtta tctgtcgctt tgagtcaaag
     3001 aactacagtt cctgaggaag aactaaatcc aaagatacta gctttgcaga atgctcagag
     3061 aaagcgaaaa atggaacatg atggttcact ttttcaagca gtaggaattg gaacattatt
     3121 acagcagcca gacgatcatg cagctactac atcactttct tggaaacgtg taaaaggatg
     3181 caaatctagt gaacagaatg gaatggagca aaagacaatt attttaatac cctctgattt
     3241 agcatgtaga ctgctggggc aatcaatgga tgaaagtgga ttaccacagc tgaccagtta
     3301 tgattgtgaa gttaatgctc ctatacaagg cagcagaaac ctactgcagg gtgaagaatt
     3361 actcagagct ttggatcaag ttaactgact cgagtctaga gggcccgttt aaacccgctg
     3421 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     3481 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     3541 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     3601 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc
     3661 tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
     3721 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     3781 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     3841 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     3901 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     3961 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     4021 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     4081 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg
     4141 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     4201 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     4261 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     4321 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     4381 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga
     4441 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt
     4501 ttcggatctg atcaagagac aggatgagga tcgtttcgca tgaccgagta caagcccacg
     4561 gtgcgcctcg ccacccgcga cgacgtcccc cgggccgtac gcaccctcgc cgccgcgttc
     4621 gccgactacc ccgccacgcg ccacaccgtc gacccggacc gccacatcga gcgggtcacc
     4681 gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg
     4741 gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg
     4801 ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa
     4861 cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc
     4921 gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga
     4981 gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac
     5041 ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga
     5101 ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgag cgggactctg gggttcgaaa
     5161 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     5221 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     5281 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     5341 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     5401 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     5461 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     5521 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     5581 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     5641 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     5701 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     5761 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     5821 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     5881 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     5941 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     6001 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     6061 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     6121 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     6181 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     6241 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     6301 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     6361 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     6421 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     6481 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     6541 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     6601 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     6661 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     6721 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     6781 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     6841 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     6901 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     6961 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     7021 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     7081 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     7141 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     7201 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     7261 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     7321 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     7381 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     7441 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     7501 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     7561 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     7621 aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt
//
