LOCUS       V018903                 7439 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018903
VERSION     V018903
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             929..2929
                     /label="HIF3A(NM_152794)"
                     /note="HIF3A(NM_152794)"
                     /gene="HIF3A"
     CDS             2936..3001
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3039..3263
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3309..3737
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3751..4080
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4147..4941
                     /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    5115..5236
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5285..5301)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5309..5325
                     /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(5333..5363)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5378..5399
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5687..6272)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6443..7303)
                     /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(7304..7408)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgccaccat ggactggcaa gaccacaggt cgaccacgga
      961 gctgcgcaag gaaaagtccc gggatgcggc ccgcagccgg cgcagccagg agaccgaggt
     1021 gctgtaccag ctggctcaca cgctgccctt cgcccgcggc gtcagcgccc acctggacaa
     1081 ggcctctatc atgcgcctca ccatcagcta cctgcgcatg caccgcctct gcgccgcagg
     1141 ggagtggaac caggtgggag cagggggaga accactggat gcctgctacc tgaaggccct
     1201 ggagggcttc gtcatggtgc tcaccgccga gggagacatg gcttacctgt cggagaatgt
     1261 cagcaaacac ctgggcctca gtcagctgga gctcattgga cacagcatct ttgatttcat
     1321 ccacccctgt gaccaagagg agcttcagga cgccctgacc ccccagcaga ccctgtccag
     1381 gaggaaggtg gaggccccca cggagcggtg cttctccttg cgcatgaaga gtacactcac
     1441 cagccgcggg cgcaccctca acctcaaggc ggccacctgg aaggtgctga actgctctgg
     1501 acatatgagg gcctacaagc cacctgcgca gacttctcca gctgggagcc ctgactcaga
     1561 gcccccgctg cagtgcctgg tgctcatctg cgaagccatc ccccacccag gcagcctgga
     1621 gcccccactg ggccgagggg ccttcctcag ccgccacagc ctggacatga agttcaccta
     1681 ctgtgacgac aggattgcag aagtggctgg ctatagtccc gatgacctga tcggctgttc
     1741 cgcctacgag tacatccacg cgctggactc cgatgcggtc agcaagagca tccacacctt
     1801 gctgagcaag ggccaggcag taacagggca gtatcgcttc ctggcccgga gtggtggcta
     1861 cctgtggacc cagacccagg ccacagtggt gtcaggggga cggggccccc agtcggagag
     1921 tatcgtctgt gtccattttt taatcagcca ggtggaagag accggagtgg tgctgtccct
     1981 ggagcaaacg gagcaacact ctcgcagacc cattcagcgg ggcgccccct ctcagaagga
     2041 cacccctaac cctggggaca gccttgacac ccctggcccc cggatccttg ccttcctgca
     2101 cccgccttcc ctgagcgagg ctgccctggc cgctgacccc cgccgtttct gcagccctga
     2161 cctccgtcgc ctcctgggac ccatcctgga tggggcttca gtagcagcca ctcccagcac
     2221 cccgctggcc acacggcacc cccaaagtcc tctttcggct gatctcccag atgaactacc
     2281 tgtgggcacc gagaatgtgc acagactctt cacctccggg aaagacactg aggcagtgga
     2341 gacagattta gatatagctc aggatgctga tgctctggat ttggagatgc tggcccccta
     2401 catctccatg gatgatgact tccagctcaa cgccagcgag cagctaccca gggcctacca
     2461 cagacctctg ggggctgtcc cccggccccg tgctcggagc ttccatggcc tgtcacctcc
     2521 agcccttgag ccctccctgc taccccgctg ggggagtgac ccccggctga gctgctccag
     2581 cccttccaga ggggacccct cagcatcctc tcccatggct ggggctcgga agaggaccct
     2641 ggcccagagc tcagaggacg aggacgaggg agtggagctg ctgggagtga gacctcccaa
     2701 aaggtccccc agcccagaac acgaaaactt tctgctcttt cctctcagcc tgagtttcct
     2761 tctgacagga ggaccagccc cagggagcct gcaggacccc agcaccccac tcctgaacct
     2821 gaatgagccc ctgggcctgg gcccctcact gctctctccg tactcagacg aggacactac
     2881 ccagcccggg ggccccttcc agccaagggc aggctcagcc caggctgacc tcgaggacta
     2941 caaagaccat gacggtgatt ataaagatca tgacatcgac tacaaggatg acgatgacaa
     3001 gtagtgaggg cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc
     3061 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     3121 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     3181 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
     3241 tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg
     3301 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     3361 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     3421 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     3481 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     3541 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     3601 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     3661 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     3721 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     3781 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     3841 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3901 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     3961 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     4021 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     4081 aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg
     4141 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     4201 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     4261 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     4321 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     4381 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     4441 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     4501 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     4561 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     4621 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     4681 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     4741 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     4801 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     4861 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     4921 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     4981 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     5041 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     5101 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     5161 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     5221 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat
     5281 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     5341 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     5401 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     5461 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     5521 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     5581 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     5641 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     5701 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     5761 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     5821 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     5881 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     5941 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     6001 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     6061 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     6121 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     6181 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggttttttt gtttgcaagc
     6241 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     6301 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     6361 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     6421 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     6481 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     6541 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     6601 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     6661 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     6721 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     6781 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     6841 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     6901 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     6961 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     7021 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     7081 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     7141 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     7201 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     7261 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     7321 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     7381 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtc
//
