LOCUS       V027329                 8629 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027329
VERSION     V027329
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             874..1590
                     /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"
     CDS             1642..4119
                     /label="HIF1A(NM_001530)"
                     /note="HIF1A(NM_001530)"
                     /gene="HIF1A"
     polyA_signal    4196..4420
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4466..4894
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4908..5237
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5304..6098
                     /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    6272..6393
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6442..6458)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6466..6482
                     /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(6490..6520)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6535..6556
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6844..7429)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7600..8460)
                     /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(8461..8565)
                     /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 ggctagcgcc accatggtga gcaagggcga ggagctgttc
      901 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
      961 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     1021 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg
     1081 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     1141 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     1201 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     1261 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     1321 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
     1381 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     1441 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc
     1501 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     1561 atcactctcg gcatggacga gctgtacaag ggaggcggag gtagtggagg agggggaagc
     1621 ggaggcggag ggagtgaatt catggagggc gccggcggcg cgaacgacaa gaaaaagata
     1681 agttctgaac gtcgaaaaga aaagtctcga gatgcagcca gatctcggcg aagtaaagaa
     1741 tctgaagttt tttatgagct tgctcatcag ttgccacttc cacataatgt gagttcgcat
     1801 cttgataagg cctctgtgat gaggcttacc atcagctatt tgcgtgtgag gaaacttctg
     1861 gatgctggtg atttggatat tgaagatgac atgaaagcac agatgaattg cttttatttg
     1921 aaagccttgg atggttttgt tatggttctc acagatgatg gtgacatgat ttacatttct
     1981 gataatgtga acaaatacat gggattaact cagtttgaac taactggaca cagtgtgttt
     2041 gattttactc atccatgtga ccatgaggaa atgagagaaa tgcttacaca cagaaatggc
     2101 cttgtgaaaa agggtaaaga acaaaacaca cagcgaagct tttttctcag aatgaagtgt
     2161 accctaacta gccgaggaag aactatgaac ataaagtctg caacatggaa ggtattgcac
     2221 tgcacaggcc acattcacgt atatgatacc aacagtaacc aacctcagtg tgggtataag
     2281 aaaccaccta tgacctgctt ggtgctgatt tgtgaaccca ttcctcaccc atcaaatatt
     2341 gaaattcctt tagatagcaa gactttcctc agtcgacaca gcctggatat gaaattttct
     2401 tattgtgatg aaagaattac cgaattgatg ggatatgagc cagaagaact tttaggccgc
     2461 tcaatttatg aatattatca tgctttggac tctgatcatc tgaccaaaac tcatcatgat
     2521 atgtttacta aaggacaagt caccacagga cagtacagga tgcttgccaa aagaggtgga
     2581 tatgtctggg ttgaaactca agcaactgtc atatataaca ccaagaattc tcaaccacag
     2641 tgcattgtat gtgtgaatta cgttgtgagt ggtattattc agcacgactt gattttctcc
     2701 cttcaacaaa cagaatgtgt ccttaaaccg gttgaatctt cagatatgaa aatgactcag
     2761 ctattcacca aagttgaatc agaagataca agtagcctct ttgacaaact taagaaggaa
     2821 cctgatgctt taactttgct ggccccagcc gctggagaca caatcatatc tttagatttt
     2881 ggcagcaacg acacagaaac tgatgaccag caacttgagg aagtaccatt atataatgat
     2941 gtaatgctcc cctcacccaa cgaaaaatta cagaatataa atttggcaat gtctccatta
     3001 cccaccgctg aaacgccaaa gccacttcga agtagtgctg accctgcact caatcaagaa
     3061 gttgcattaa aattagaacc aaatccagag tcactggaac tttcttttac catgccccag
     3121 attcaggatc agacacctag tccttccgat ggaagcacta gacaaagttc acctgagcct
     3181 aatagtccca gtgaatattg tttttatgtg gatagtgata tggtcaatga attcaagttg
     3241 gaattggtag aaaaactttt tgctgaagac acagaagcaa agaacccatt ttctactcag
     3301 gacacagatt tagacttgga gatgttagct ccctatatcc caatggatga tgacttccag
     3361 ttacgttcct tcgatcagtt gtcaccatta gaaagcagtt ccgcaagccc tgaaagcgca
     3421 agtcctcaaa gcacagttac agtattccag cagactcaaa tacaagaacc tactgctaat
     3481 gccaccacta ccactgccac cactgatgaa ttaaaaacag tgacaaaaga ccgtatggaa
     3541 gacattaaaa tattgattgc atctccatct cctacccaca tacataaaga aactactagt
     3601 gccacatcat caccatatag agatactcaa agtcggacag cctcaccaaa cagagcagga
     3661 aaaggagtca tagaacagac agaaaaatct catccaagaa gccctaacgt gttatctgtc
     3721 gctttgagtc aaagaactac agttcctgag gaagaactaa atccaaagat actagctttg
     3781 cagaatgctc agagaaagcg aaaaatggaa catgatggtt cactttttca agcagtagga
     3841 attggaacat tattacagca gccagacgat catgcagcta ctacatcact ttcttggaaa
     3901 cgtgtaaaag gatgcaaatc tagtgaacag aatggaatgg agcaaaagac aattatttta
     3961 ataccctctg atttagcatg tagactgctg gggcaatcaa tggatgaaag tggattacca
     4021 cagctgacca gttatgattg tgaagttaat gctcctatac aaggcagcag aaacctactg
     4081 cagggtgaag aattactcag agctttggat caagttaact gaggatcctg atatccagca
     4141 cagtggcggc cgctcgagtc tagagggccc gtttaaaccc gctgatcagc ctcgactgtg
     4201 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     4261 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     4321 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     4381 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc
     4441 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     4501 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     4561 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     4621 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     4681 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     4741 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     4801 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     4861 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
     4921 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4981 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     5041 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     5101 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     5161 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     5221 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag
     5281 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     5341 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     5401 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     5461 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
     5521 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     5581 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     5641 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     5701 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     5761 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     5821 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     5881 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     5941 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     6001 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     6061 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     6121 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     6181 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     6241 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     6301 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     6361 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     6421 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     6481 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     6541 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     6601 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     6661 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     6721 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     6781 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     6841 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     6901 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     6961 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     7021 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     7081 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     7141 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     7201 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     7261 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     7321 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     7381 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     7441 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     7501 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     7561 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     7621 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     7681 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     7741 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     7801 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     7861 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     7921 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     7981 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     8041 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     8101 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     8161 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     8221 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     8281 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     8341 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     8401 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     8461 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     8521 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     8581 agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggt
//
