LOCUS       V043792                 7709 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V043792
VERSION     V043792
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..3199
                     /label="DOK7(NM_173660)"
                     /note="DOK7(NM_173660)"
                     /gene="DOK7"
     CDS             3206..3271
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3309..3533
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3579..4007
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4021..4350
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4417..5211
                     /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    5385..5506
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5555..5571)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5579..5595
                     /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(5603..5633)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5648..5669
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5957..6542)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6713..7573)
                     /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(7574..7678)
                     /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 accgaggcgg cgctggtgga gggccaggtc aagctgcggg acggcaagaa
     1741 gtggaagagt aggtggctgg tgctgcggaa gccgtcgccc gtggcagact gcctgctgat
     1801 gctggtctac aaggacaagt cggagcgtat caagggcctg cgggagcgca gcagcctgac
     1861 gctagaggac atctgcgggc tggagcccgg cctgccctac gagggcctgg tccacacgct
     1921 ggccattgtc tgcctgtccc aggccatcat gctgggcttt gacagccacg aggccatgtg
     1981 tgcgtgggat gcccggatcc gctatgcgct cggcgaggtg cataggttcc atgtgacagt
     2041 ggctccaggc accaagttgg agagcggccc ggctaccctg cacctctgca atgatgtcct
     2101 cgtcttggcc agggacatcc ccccggctgt cacggggcag tggaagctgt ctgacctccg
     2161 gcgctacggg gccgtgccaa gcggattcat ctttgaaggc gggaccaggt gtgggtactg
     2221 ggctggcgtc ttcttcctgt cctcggccga gggggagcag atcagcttcc tgttcgactg
     2281 catcgtccga ggcatctccc ccaccaaggg cccctttggg ctgcggccgg ttctaccaga
     2341 cccaagtccc ccgggaccct cgactgtgga ggagcgtgtg gcccaggaag ccctggaaac
     2401 cctacagctg gagaagcggc tgagcctcct ctcacatgcg ggcaggccgg gcagtggagg
     2461 ggatgaccgc agcctgtcca gctcatcctc agaggccagt cacttggacg tcagcgccag
     2521 cagccggctc accgcatggc cagagcaatc ctcgtcgtca gccagcacgt cacaggaggg
     2581 gcctagacca gcagctgccc aggccgccgg ggaagccatg gtgggtgcct caaggccacc
     2641 ccccaagccg ctgcgtccgc ggcagctgca ggaggttggc cgccagagct cctcggacag
     2701 cggcatcgcc actggcagcc actcctctta ctccagcagc ctctcgtcct acgcgggcag
     2761 cagcctggac gtgtggcggg ccacagatga actgggctca ctgctcagcc tgccagcagc
     2821 gggggccccc gagcccagcc tgtgcacctg cctgcccggg acagtcgagt accaggtgcc
     2881 cacctccctg cgggcccact atgacacacc acgcagcctt tgcctggctc ctagagacca
     2941 cagccccccc tcacagggca gccccggcaa cagtgcggcc agggactcag gcggccagac
     3001 gtccgccggg tgtccctctg gctggctggg cacgagacgg cggggcctgg tgatggaggc
     3061 cccccagggc agcgaggcca cactgcctgg ccctgcccct ggcgagccct gggaagcagg
     3121 cggcccccac gcggggccac ccccggcttt cttttcggca tgtccagtct gtggaggact
     3181 caaggtaaac ccccctcctc tcgaggacta caaagaccat gacggtgatt ataaagatca
     3241 tgacatcgac tacaaggatg acgatgacaa gtagtgaggg cccgtttaaa cccgctgatc
     3301 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     3361 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     3421 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     3481 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga
     3541 ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
     3601 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     3661 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     3721 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     3781 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     3841 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     3901 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     3961 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
     4021 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4081 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     4141 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     4201 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     4261 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
     4321 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
     4381 ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     4441 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     4501 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     4561 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     4621 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     4681 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     4741 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     4801 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     4861 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     4921 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     4981 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     5041 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     5101 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     5161 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     5221 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     5281 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     5341 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag
     5401 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     5461 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac
     5521 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     5581 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     5641 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     5701 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     5761 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     5821 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     5881 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     5941 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     6001 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     6061 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     6121 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     6181 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     6241 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     6301 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     6361 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     6421 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     6481 ccgctggtag cggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     6541 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     6601 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     6661 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     6721 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     6781 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     6841 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     6901 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     6961 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     7021 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     7081 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     7141 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     7201 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     7261 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     7321 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     7381 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     7441 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     7501 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     7561 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     7621 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     7681 catttccccg aaaagtgcca cctgacgtc
//
