LOCUS       V037858                10423 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V037858
VERSION     V037858
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             4489..5913
                     /label="GABRG2(NM_198904)"
                     /note="GABRG2(NM_198904)"
                     /gene="GABRG2"
     polyA_signal    5990..6214
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      6260..6688
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        6702..7031
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             7098..7892
                     /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    8066..8187
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(8236..8252)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    8260..8276
                     /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(8284..8314)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    8329..8350
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8638..9223)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(9394..10254)
                     /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(10255..10359)
                     /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 catgtggggc cttgcgggag gaaggctttt cggcatcttc
     1681 tcggccccgg tgctggtggc tgtggtgtgc tgcgcccaga gtgtgaacga tcccgggaac
     1741 atgtcctttg tgaaggagac ggtggacaag ctgttgaaag gctacgacat tcgcctaaga
     1801 cccgacttcg ggggtccccc ggtctgcgtg gggatgaaca tcgacatcgc cagcatcgac
     1861 atggtttccg aagtcaacat ggattatacc ttaaccatgt attttcaaca atattggaga
     1921 gataaaaggc tcgcctattc tgggatccct ctcaacctca cgcttgacaa tcgagtggct
     1981 gaccagctat gggtgcccga cacatatttc ttaaatgaca aaaagtcatt tgtgcatgga
     2041 gtgacagtga aaaaccgcat gatccgtctt caccctgatg ggacagtgct gtatgggctc
     2101 agaatcacca cgacagcagc atgcatgatg gacctcagga gataccccct ggacgagcag
     2161 aactgcactc tggaaattga aagctatggc tacaccacgg atgacattga gttttactgg
     2221 cgaggcgggg acaaggctgt taccggagtg gaaaggattg agctcccgca gttctccatc
     2281 gtggagcacc gtctggtctc gaggaatgtt gtcttcgcca caggtgccta tcctcgactg
     2341 tcactgagct ttcggttgaa gaggaacatt ggatacttca ttcttcagac ttatatgccc
     2401 tctatactga taacgattct gtcgtgggtg tccttctgga tcaattatga tgcatctgct
     2461 gctagagttg ccctcgggat cacaactgtg ctgacaatga caaccatcaa cacccacctt
     2521 cgggagacct tgcccaaaat cccctatgtc aaagccattg acatgtacct tatgggctgc
     2581 ttcgtctttg tgttcctggc ccttctggag tatgcctttg tcaactacat tttctttgga
     2641 agaggccctc aaaggcagaa gaagcttgca gaaaagacag ccaaggcaaa gaatgaccgt
     2701 tcaaagagcg aaagcaaccg ggtggatgct catggaaata ttctgttgac atcgctggaa
     2761 gttcacaatg aaatgaatga ggtctcaggc ggcattggcg ataccaggaa ttcagcaata
     2821 tcctttgaca actcaggaat ccagtacagg aaacagagca tgcctcgaga agggcatggg
     2881 cgattcctgg gggacagaag cctcccgcac aagaagaccc atctacggag gaggtcttca
     2941 cagctcaaaa ttaaaatacc tgatctaacc gatgtgaatg ccatagacag atggtccagg
     3001 atcgtgtttc cattcacttt ttctcttttc aacttagttt actggctgta ctatgttaac
     3061 ggaggtggag gtagtggagg aggcggatca atgaggaaaa gtccaggtct gtctgactgt
     3121 ctttgggcct ggatcctcct tctgagcaca ctgactggaa gaagctatgg acagccgtca
     3181 ttacaagatg aacttaaaga caataccact gtcttcacca ggattttgga cagactccta
     3241 gatggttatg acaatcgcct gagaccagga ttgggagagc gtgtaaccga agtgaagact
     3301 gatatcttcg tcaccagttt cggacccgtt tcagaccatg atatggaata tacaatagat
     3361 gtatttttcc gtcaaagctg gaaggatgaa aggttaaaat ttaaaggacc tatgacagtc
     3421 ctccggttaa ataacctaat ggcaagtaaa atctggactc cggacacatt tttccacaat
     3481 ggaaagaagt cagtggccca caacatgacc atgcccaaca aactcctgcg gatcacagag
     3541 gatggcacct tgctgtacac catgaggctg acagtgagag ctgaatgtcc gatgcatttg
     3601 gaggacttcc ctatggatgc ccatgcttgc ccactaaaat ttggaagtta tgcttataca
     3661 agagcagaag ttgtttatga atggaccaga gagccagcac gctcagtggt tgtagcagaa
     3721 gatggatcac gtctaaacca gtatgacctt cttggacaaa cagtagactc tggaattgtc
     3781 cagtcaagta caggagaata tgttgttatg accactcatt tccacttgaa gagaaagatt
     3841 ggctactttg ttattcaaac atacctgcca tgcataatga cagtgattct ctcacaagtc
     3901 tccttctggc tcaacagaga gtctgtacca gcaagaactg tctttggagt aacaactgtg
     3961 ctcaccatga caacattgag catcagtgcc agaaactccc tccctaaggt ggcttatgca
     4021 acagctatgg attggtttat tgccgtgtgc tatgcctttg tgttctcagc tctgattgag
     4081 tttgccacag taaactattt cactaagaga ggttatgcat gggatggcaa aagtgtggtt
     4141 ccagaaaagc caaagaaagt aaaggatcct cttattaaga aaaacaacac ttacgctcca
     4201 acagcaacca gctacacccc taatttggcc aggggcgacc cgggcttagc caccattgct
     4261 aaaagtgcaa ccatagaacc taaagaggtc aagcccgaaa caaaaccacc agaacccaag
     4321 aaaaccttta acagtgtcag caaaattgac cgactgtcaa gaatagcctt cccgctgcta
     4381 tttggaatct ttaacttagt ctactgggct acgtatttaa acagagagcc tcagctaaaa
     4441 gcccccacac cacatcaagg cggtggaggc agtggtggag gcggtagcat gagttcgcca
     4501 aatatatgga gcacaggaag ctcagtctac tcgactcctg tattttcaca gaaaatgacg
     4561 gtgtggattc tgctcctgct gtcgctctac cctggcttca ctagccagaa atctgatgat
     4621 gactatgaag attatgcttc taacaaaaca tgggtcttga ctccaaaagt tcctgagggt
     4681 gatgtcactg tcatcttaaa caacctgctg gaaggatatg acaataaact tcggcctgat
     4741 ataggagtga agccaacgtt aattcacaca gacatgtatg tgaatagcat tggtccagtg
     4801 aacgctatca atatggaata cactattgat atattttttg cgcaaacgtg gtatgacaga
     4861 cgtttgaaat ttaacagcac cattaaagtc ctccgattga acagcaacat ggtggggaaa
     4921 atctggattc cagacacttt cttcagaaat tccaaaaaag ctgatgcaca ctggatcacc
     4981 acccccaaca ggatgctgag aatttggaat gatggtcgag tgctctacac cctaaggttg
     5041 acaattgatg ctgagtgcca attacaattg cacaactttc caatggatga acactcctgc
     5101 cccttggagt tctccagtta tggctatcca cgtgaagaaa ttgtttatca atggaagcga
     5161 agttctgttg aagtgggcga cacaagatcc tggaggcttt atcaattctc atttgttggt
     5221 ctaagaaata ccaccgaagt agtgaagaca acttccggag attatgtggt catgtctgtc
     5281 tactttgatc tgagcagaag aatgggatac tttaccatcc agacctatat cccctgcaca
     5341 ctcattgtcg tcctatcctg ggtgtctttc tggatcaata aggatgctgt tccagccaga
     5401 acatctttag gtatcaccac tgtcctgaca atgaccaccc tcagcaccat tgcccggaaa
     5461 tcgctcccca aggtctccta tgtcacagcg atggatctct ttgtatctgt ttgtttcatc
     5521 tttgtcttct ctgctctggt ggagtatggc accttgcatt attttgtcag caaccggaaa
     5581 ccaagcaagg acaaagataa aaagaagaaa aaccctcttc ttcggatgtt ttccttcaag
     5641 gcccctacca ttgatatccg cccaagatca gcaaccattc aaatgaataa tgctacacac
     5701 cttcaagaga gagatgaaga gtacggctat gagtgtctgg acggcaagga ctgtgccagt
     5761 tttttctgct gttttgaaga ttgtcgaaca ggagcttgga gacatgggag gatacatatc
     5821 cgcattgcca aaatggactc ctatgctcgg atcttcttcc ccactgcctt ctgcctgttt
     5881 aatctggtct attgggtctc ctacctctac ctgtgaggat cctgatatcc agcacagtgg
     5941 cggccgctcg agtctagagg gcccgtttaa acccgctgat cagcctcgac tgtgccttct
     6001 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     6061 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     6121 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     6181 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
     6241 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     6301 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     6361 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     6421 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     6481 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     6541 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     6601 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     6661 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg
     6721 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     6781 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     6841 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     6901 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     6961 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     7021 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag
     7081 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     7141 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     7201 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     7261 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     7321 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     7381 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     7441 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     7501 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     7561 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     7621 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     7681 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     7741 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     7801 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     7861 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     7921 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     7981 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     8041 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     8101 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     8161 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     8221 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     8281 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     8341 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     8401 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     8461 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     8521 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     8581 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     8641 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     8701 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     8761 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     8821 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     8881 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     8941 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     9001 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     9061 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     9121 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtttttt
     9181 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     9241 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     9301 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     9361 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     9421 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     9481 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     9541 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     9601 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     9661 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     9721 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     9781 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     9841 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     9901 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     9961 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
    10021 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
    10081 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
    10141 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
    10201 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
    10261 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
    10321 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
    10381 acctgacgtc gacggatcgg gagatctccc gatcccctat ggt
//
