LOCUS       V030515                 9383 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V030515
VERSION     V030515
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             1666..4755
                     /label="PPP1R12A(NM_002480)"
                     /note="PPP1R12A(NM_002480)"
                     /gene="PPP1R12A"
     CDS             4762..4788
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             4792..4818
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             4822..4848
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     polyA_signal    4950..5174
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      5220..5648
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5662..5991
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6058..6852
                     /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    7026..7147
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7196..7212)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7220..7236
                     /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(7244..7274)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7289..7310
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7598..8183)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8354..9214)
                     /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(9215..9319)
                     /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 tccggactca gatctcgagg cggcggaggg
     1621 agtggaggag ggggaagcgg aggcggaggg agtaagcttg ccaccatgaa gatggcggac
     1681 gcgaagcaga agcggaacga gcagctgaaa cgctggatcg gctccgagac ggacctcgag
     1741 cctccggtgg tgaagcgcca gaagaccaag gtgaagttcg acgatggcgc cgtcttcctg
     1801 gctgcttgct ccagcggcga cacggacgag gtcctcaagc tgctgcaccg cggcgccgac
     1861 atcaattacg ccaatgtgga cggactcact gccctgcacc aggcttgcat tgatgacaat
     1921 gttgatatgg tgaagtttct ggtagaaaat ggagcaaata ttaatcaacc tgataatgaa
     1981 ggctggatac cactacatgc agcagcttcc tgtggatatc ttgatattgc agagtttttg
     2041 attggtcaag gagcacatgt aggggctgtc aacagtgaag gagatacacc tttagatatt
     2101 gcggaggagg aggcaatgga agagctactt caaaatgaag ttaatcggca aggggttgat
     2161 atagaagcag ctcgaaagga agaagaacgg atcatgctta gagatgccag gcagtggcta
     2221 aatagtggtc atataaatga tgtccggcat gcaaaatctg gaggtacagc acttcacgtt
     2281 gcagctgcta aaggctatac ggaagtttta aaacttttaa tacaggcagg ctatgatgtt
     2341 aatattaaag actatgatgg ctggacacct cttcatgctg cagctcattg gggtaaagaa
     2401 gaagcatgtc gaattttagt ggacaatctg tgtgatatgg agatggtcaa caaagtgggc
     2461 caaacagcct ttgatgtagc agatgaagac attttaggat atttagaaga gttgcaaaag
     2521 aaacaaaatc tgctccatag tgaaaaacgg gacaagaaat ctccactaat tgaatcaaca
     2581 gcaaatatgg acaataatca gtcacagaag acctttaaaa acaaagagac gttgattatt
     2641 gaaccagaga aaaatgcatc ccgtattgaa tctctggaac aagaaaaggt tgatgaagaa
     2701 gaagaaggaa agaaggatga gtctagctgc tctagtgaag aagatgagga agatgactcg
     2761 gaatcagaag ctgaaacaga taagacaaaa cccctggctt ctgtaactaa tgccaacact
     2821 tctagtacac aagcagctcc tgtagctgtt acaacaccta ctgtgtcatc aggtcaagca
     2881 acacctacat cacctattaa aaagtttcca accacagcta caaaaatttc tcccaaagaa
     2941 gaagagagaa aagatgagtc tcctgcaact tggaggttag gacttagaaa gacgggcagc
     3001 tatggtgcac ttgctgaaat cacagcatct aaagagggtc agaaagaaaa agatactgca
     3061 ggtgttacac gttcagcttc aagtcccaga ctttcctcct ctttggataa taaagaaaag
     3121 gagaaagata gtaaaggaac taggcttgca tatgttgcac ctacaatacc aagacgacta
     3181 gccagtacat ctgacattga agagaaagaa aacagagatt cttcaagttt gcgaacaagt
     3241 agttcatata caaggagaaa atgggaagat gatcttaaaa aaaatagctc agttaatgaa
     3301 ggatcaacgt atcataaaag ttgctccttt ggtagaagac aagatgattt gattagttct
     3361 agtgttccaa gcaccacatc aacaccaaca gttacctctg cagctgggct tcagaaaagc
     3421 ctgctttcca gcacaagcac tactacaaag attacaacgg gttcttcctc agcaggcaca
     3481 caaagcagta cctcaaatcg tttgtgggct gaggatagta ctgagaaaga aaaggacagt
     3541 gttcctacgg cagtgaccat tcctgttgct ccaactgttg taaatgctgc agcttctacc
     3601 acaaccctga ctacaactac tgctggcact gtctcctcca caacagaggt cagggagaga
     3661 cgcagatcat acctcactcc tgttagggat gaagagtctg aatcccaaag aaaagcaaga
     3721 tctagacaag caagacaatc tagaagatca acacagggag tgacattaac tgatcttcaa
     3781 gaagctgaga aaacaatagg aagaagtcgt tctacccgaa ccagagaaca agaaaatgaa
     3841 gaaaaagaaa aagaggaaaa agagaaacaa gataaagaga aacaagaaga aaagaaggag
     3901 tcagaaacat ctagagaaga tgaatataaa caaaagtact ccagaacgta tgatgagact
     3961 taccagcgtt ataggccagt atcaacttca agttcaacca ctccatcctc ttcactttct
     4021 actatgagca gttcactgta tgcttcaagt caactaaaca ggccaaatag tcttgtaggc
     4081 ataacttctg cttactccag aggaataaca aaagaaaatg aaagagaggg agaaaaaaga
     4141 gaagaggaga aagaaggaga agataaatca caacctaaat caatcagaga acgacgacga
     4201 ccaagagaga aaagaagatc tacaggagtt tcattttgga cacaagatag tgatgaaaat
     4261 gaacaagaac aacaatcaga cacagaagag ggatccaata agaaagaaac tcagacggat
     4321 tccatttcta gatatgaaac cagttctaca tcagctggtg atcgatatga ttccttgctg
     4381 ggtcgctctg gatcatacag ttacttagaa gaaagaaaac cttacagcag caggctagaa
     4441 aaggatgact caactgactt taaaaagctt tatgaacaaa ttctagctga aaatgaaaag
     4501 ctgaaggcac agctacatga tacaaatatg gaactaacag atcttaaatt acagttggaa
     4561 aaggccaccc agagacaaga aagatttgct gatagatcac tgttggaaat ggaaaaaagg
     4621 gaacgaagag ctctagaaag aagaatatct gaaatggaag aagagctcaa aatgttacca
     4681 gacctaaaag cagacaacca gaggctaaag gatgaaaatg gggccttgat cagagttata
     4741 agcaaacttt ccaaaggtac ctacccatac gacgtacctg attatgcaac ttatccttat
     4801 gatgtacctg attatgctag ctatccttat gatgtcccag actacgcata ggatccacta
     4861 gtccagtgtg gtggaattct gcagatatcc agcacagtgg cggccgctcg agtctagagg
     4921 gcccgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     4981 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     5041 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     5101 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
     5161 ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca
     5221 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     5281 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     5341 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     5401 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     5461 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     5521 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     5581 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     5641 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     5701 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     5761 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     5821 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     5881 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     5941 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     6001 ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg
     6061 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     6121 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     6181 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     6241 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     6301 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     6361 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     6421 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     6481 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     6541 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     6601 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     6661 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     6721 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     6781 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     6841 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     6901 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     6961 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     7021 accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     7081 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     7141 tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
     7201 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     7261 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     7321 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     7381 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     7441 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     7501 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     7561 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     7621 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     7681 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     7741 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     7801 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     7861 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     7921 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     7981 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     8041 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     8101 cttgatccgg caaacaaacc accgctggta gcggtttttt tgtttgcaag cagcagatta
     8161 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     8221 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     8281 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     8341 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     8401 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     8461 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     8521 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     8581 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     8641 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     8701 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     8761 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     8821 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     8881 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     8941 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     9001 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     9061 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     9121 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     9181 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     9241 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     9301 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg
     9361 gagatctccc gatcccctat ggt
//
