LOCUS       V035138                 7865 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V035138
VERSION     V035138
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..5334
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    5508..5629
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5678..5694)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5702..5718
                     /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(5726..5756)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5771..5792
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6080..6665)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6836..7696)
                     /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(7697..7801)
                     /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 tgattgaaca agatggattg
     4561 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     4621 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     4681 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta
     4741 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     4801 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     4861 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     4921 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     4981 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     5041 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc
     5101 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     5161 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     5221 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     5281 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     5341 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
     5401 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     5461 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg
     5521 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     5581 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta
     5641 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     5701 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     5761 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     5821 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     5881 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     5941 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     6001 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     6061 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     6121 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     6181 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     6241 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     6301 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     6361 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     6421 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     6481 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     6541 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     6601 ccaccgctgg tagcggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     6661 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     6721 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     6781 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     6841 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     6901 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     6961 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     7021 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     7081 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     7141 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     7201 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     7261 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     7321 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     7381 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     7441 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     7501 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     7561 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     7621 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     7681 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     7741 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     7801 gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct
     7861 atggt
//
