LOCUS       V027567                 7550 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027567
VERSION     V027567
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             935..1651
                     /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             1688..3040
                     /label="PTK6(NM_005975)"
                     /note="PTK6(NM_005975)"
                     /gene="PTK6"
     CDS             3047..3112
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3150..3374
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3420..3848
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3862..4191
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4258..5052
                     /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    5226..5347
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5396..5412)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5420..5436
                     /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(5444..5474)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5489..5510
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5798..6383)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6554..7414)
                     /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(7415..7519)
                     /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 ccgagctcgg atccatggtg agcaagggcg aggagctgtt
      961 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
     1021 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg
     1081 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt
     1141 gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     1201 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
     1261 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     1321 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca
     1381 caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg
     1441 ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat
     1501 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag
     1561 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
     1621 gatcactctc ggcatggacg agctgtacaa ggaattcggt ggaggcggat caggaggcgg
     1681 tggatctatg gtgtcccggg accaggctca cctgggcccc aagtatgtgg gcctctggga
     1741 cttcaagtcc cggacggacg aggagctgag cttccgcgcg ggggacgtct tccacgtggc
     1801 caggaaggag gagcagtggt ggtgggccac gctgctggac gaggcgggtg gggccgtggc
     1861 ccagggctat gtgccccaca actacctggc cgagagggag acggtggagt cggaaccgtg
     1921 gttctttggc tgcatctccc gctcggaagc tgtgcgtcgg ctgcaggccg agggcaacgc
     1981 cacgggcgcc ttcctgatca gggtcagcga gaagccgagt gccgactacg tcctgtcggt
     2041 gcgggacacg caggctgtgc ggcactacaa gatctggcgg cgtgccgggg gccggctgca
     2101 cctgaacgag gcggtgtcct tcctcagcct gcccgagctt gtgaactacc acagggccca
     2161 gagcctgtcc cacggcctgc ggctggccgc gccctgccgg aagcacgagc ctgagcccct
     2221 gccccattgg gatgactggg agaggccgag ggaggagttc acgctctgca ggaagctggg
     2281 gtccggctac tttggggagg tcttcgaggg gctctggaaa gaccgggtcc aggtggccat
     2341 taaggtgatt tctcgagaca acctcctgca ccagcagatg ctgcagtcgg agatccaggc
     2401 catgaagaag ctgcggcaca aacacatcct ggcgctgtac gccgtggtgt ccgtggggga
     2461 ccccgtgtac atcatcacgg agctcatggc caagggcagc ctgctggagc tgctccgcga
     2521 ctctgatgag aaagtcctgc ccgtttcgga gctgctggac atcgcctggc aggtggctga
     2581 gggcatgtgt tacctggagt cgcagaatta catccaccgg gacctggccg ccaggaacat
     2641 cctcgtcggg gaaaacaccc tctgcaaagt tggggacttc gggttagcca ggcttatcaa
     2701 ggaggacgtc tacctctccc atgaccacaa tatcccctac aagtggacgg cccctgaagc
     2761 gctctcccga ggccattact ccaccaaatc cgacgtctgg tcctttggga ttctcctgca
     2821 tgagatgttc agcaggggtc aggtgcccta cccaggcatg tccaaccatg aggccttcct
     2881 gagggtggac gccggctacc gcatgccctg ccctctggag tgcccgccca gcgtgcacaa
     2941 gctgatgctg acatgctggt gcagggaccc cgagcagaga ccctgcttca aggccctgcg
     3001 ggagaggctc tccagcttca ccagctacga gaacccgacc ctcgaggact acaaagacca
     3061 tgacggtgat tataaagatc atgacatcga ctacaaggat gacgatgaca agtagtgagg
     3121 gcccgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     3181 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     3241 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     3301 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
     3361 ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca
     3421 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     3481 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     3541 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     3601 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     3661 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     3721 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     3781 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     3841 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     3901 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     3961 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     4021 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     4081 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     4141 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     4201 ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg
     4261 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     4321 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     4381 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     4441 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     4501 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     4561 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     4621 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     4681 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     4741 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     4801 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     4861 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     4921 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     4981 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     5041 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     5101 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     5161 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     5221 accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     5281 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     5341 tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
     5401 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     5461 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     5521 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     5581 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     5641 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     5701 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     5761 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     5821 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     5881 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     5941 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     6001 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     6061 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     6121 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     6181 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     6241 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     6301 cttgatccgg caaacaaacc accgctggta gcggtttttt tgtttgcaag cagcagatta
     6361 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     6421 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     6481 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     6541 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     6601 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     6661 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     6721 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     6781 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     6841 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     6901 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     6961 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     7021 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     7081 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     7141 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     7201 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     7261 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     7321 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     7381 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     7441 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     7501 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
//
