LOCUS       V039272                 9093 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V039272
VERSION     V039272
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     misc_signal     190..340
                     /note="packaging signal for adenovirus serotype 5"
                     /label="Ad5 Ψ"
     promoter        469..992
                     /label="mCMV promoter"
     CDS             999..1715
                     /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"
     polyA_signal    1862..1983
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     enhancer        1991..2370
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        2371..2574
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        2619..2637
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             2712..5876
                     /label="Ptk2(NM_013081)"
                     /note="Ptk2(NM_013081)"
                     /gene="Ptk2"
     CDS             5877..5942
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    5978..6202
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     polyA_signal    6216..6350
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     protein_bind    complement(6370..6403)
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core
                     sequence (ATGTATGC) (Shaw et al., 2021)."
                     /label="loxP"
     primer_bind     complement(6425..6441)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6449..6465
                     /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(6473..6503)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6518..6539
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6827..7415)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7586..8446)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     promoter        complement(8447..8551)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 ttcatcaata atatacctta ttttggattg aagccaatat gataatgagg gggtggagtt
       61 tgtgacgtgg cgcggggcgt gggaacgggg cgggtgacgt agtagtgtgg cggaagtgtg
      121 atgttgcaag tgtggcggaa cacatgtaag cgacggatgt ggcaaaagtg acgtttttgg
      181 tgtgcgccgg tgtacacagg aagtgacaat tttcgcgcgg ttttaggcgg atgttgtagt
      241 aaatttgggc gtaaccgagt aagatttggc cattttcgcg ggaaaactga ataagaggaa
      301 gtgaaatctg aataattttg tgttactcat agcgcgtaat atttgtctag ggccgcgggg
      361 actttgaccg tttacgtgga gactcgccca ggtgtttttc tcaggtgttt tccgcgttcc
      421 gggtcaaagt tggcgtttta ttattatagt cagtctagag atatactgag tcattaggga
      481 ctttccaatg ggttttgccc agtacataag gtcaataggg gtgaatcaac aggaaagtcc
      541 cattggagcc aagtacactg agtcaatagg gactttccat tgggttttgc ccagtacaaa
      601 aggtcaatag ggggtgagtc aatgggtttt tcccattatt ggcacgtaca taaggtcaat
      661 aggggtgagt cattgggttt ttccagccaa tttaattaaa acgccatgta ctttcccacc
      721 attgacgtca atgggctatt gaaactaatg caacgtgacc tttaaacggt actttcccat
      781 agctgattaa tgggaaagta ccgttctcga gccaatacac gtcaatggga agtgaaaggg
      841 cagccaaaac gtaacaccgc cccggttttc ccctggaaat tccatattgg cacgcattct
      901 attggctgag ctgcgttcta cgtgggtata agaggcgcga ccagcgtcgg taccgtcgca
      961 gtcttcggtc tgaccaccgt agaacgcaga tcgaattcat ggtgagcaag ggcgaggagc
     1021 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
     1081 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca
     1141 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg
     1201 gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
     1261 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
     1321 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
     1381 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca
     1441 gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga
     1501 tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc
     1561 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc
     1621 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
     1681 ccgggatcac tctcggcatg gacgagctgt acaagtccgg actcagatct cgagatccac
     1741 cggatctaga taactgatca taatcagcca taccacattt gtagaggttt tacttgcttt
     1801 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
     1861 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     1921 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     1981 ttaacgcgtt gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta
     2041 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
     2101 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
     2161 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
     2221 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
     2281 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
     2341 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
     2401 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
     2461 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
     2521 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg
     2581 ctaactagag aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag
     2641 acccaagctg gctagcgttt aaacgggccc tctagactcg agcggccgcc actgtgctgg
     2701 atatcgccac catggcagct gcttatcttg acccaaactt gaatcacaca ccaagttcga
     2761 gtactaagac tcacctgggt actggcacgg aacggtcccc tggtgcaatg gaacgagtat
     2821 taaaggtctt tcattatttt gaaagcagta atgagccaac cacctgggcc agtattatca
     2881 ggcatggaga cgctactgac gtcaggggaa tcattcagaa gatagtggac agtcacaaag
     2941 taaagcacgt ggcctgctat ggatttcgcc tcagtcacct gcgatcggag gaggtgcact
     3001 ggctgcatgt ggacatgggt gtctccagtg tgagggagaa atacgaactc gcccacccac
     3061 cagaggagtg gaaatatgag ttgagaattc gttatttgcc aaaaggattt ctaaaccagt
     3121 ttactgaaga taagccaaca ttgaatttct tctatcaaca ggtgaagagt gactacatgc
     3181 tagaaatagc tgatcaagta gaccaagaca tagctttgaa gttgggttgt ttggaaatta
     3241 ggcgatccta ctgggagatg aggggcaacg ccctagagaa gaagtccaac tatgaagtgt
     3301 tagagaaaga tgttggttta aagcgatttt ttcccaagag tttactggat tccgtcaagg
     3361 ccaaaacact aagaaagctg atccagcaaa cattcaggca gtttgccaac cttaatagag
     3421 aagaaagcat cctgaaattc tttgagattc tctctccagt gtacagattt gacaaagagt
     3481 gcttcaagtg tgcccttggg tcaagttgga ttatttctgt ggaactggcg atcggcccag
     3541 aagaagggat cagttacctg acagacaagg gctgcaatcc cacacaccta gcggacttca
     3601 accaagtgca gactatccag tactcaaaca gtgaagacaa agacaggaaa ggaatgctac
     3661 aactcaaaat cgctggtgct cctgagcctc tgacagtgac ggccccatcc ctaaccattg
     3721 cagagaacat ggctgacctg atcgatggat actgccggct ggtgaacgga gccacgcagt
     3781 ccttcatcat cagaccccag aaagaaggtg aacgggcatt gccatcaata ccaaagttgg
     3841 ccaacaatga aaagcaaggc atgcggacac atgcagtctc tgtgtcagag acagatgact
     3901 atgccgagat catcgatgag gaagacacat acaccatgcc ctcaaccagg gattatgaga
     3961 ttcagagaga aagaatagaa cttggacgct gtattggaga aggccagttt ggagatgtac
     4021 atcaaggcgt gtacctgagc ccagagaatc cagctttggc tgttgcaatc aaaacatgta
     4081 aaaactgtac ttcggacagc gtgagagaga agttccttca agaagcctta acaatgcgcc
     4141 agtttgacca tcctcacatc gtgaagctga ttggagtcat tacagagaac ccagtttgga
     4201 taatcatgga gttgtgcaca cttggagagc tgaggtcatt tttgcaagta aggaaataca
     4261 gcttggatct agcatctttg atcttatatg cctatcagct tagtacagca ctcgcgtatc
     4321 tggagagcaa aagatttgtt cacagggaca ttgctgcacg gaatgttctg gtgtcttcaa
     4381 atgattgtgt aaaattagga gactttggat tatctcgata tatggaagac agtacttact
     4441 ataaagcttc caaaggaaaa ttacctatta aatggatggc tccagagtca atcaattttc
     4501 gacgttttac ctcagctagt gacgtgtgga tgtttggtgt gtgtatgtgg gagatactga
     4561 tgcatggcgt gaagcctttt caaggagtga agaacaatga tgtgatcggt cgaattgaaa
     4621 atggggaaag attaccaatg cctccaaatt gtcctcccac cctctacagc cttatgacga
     4681 aatgttgggc ctatgacccc agcagacggc ccaggtttac tgaactaaaa gctcagctca
     4741 gcacaatcct ggaggaagag aaggtgcagc aagaagaacg gatgaggatg gaatccagaa
     4801 gacaggcaac agtgtcctgg gactctggag ggtctgatga agcaccaccc aagcccagca
     4861 gacctggtta tcctagcccg aggtccagtg aaggatttta tcccagccca cagcacatgg
     4921 tccagaccaa tcactatcag atctctggct acccaggttc ccatggaatc ccagccatgg
     4981 ctggcagcat ttacccaggt caggcatctc ttctggacca gacagaattg tggaatcaca
     5041 gacctcagga gatgtccatg tggcagccca gtgtggagga ctcagcagct ttagatcttc
     5101 gagggatggg acaggtgctt cctcctcacc tgatggaaga gcggctaatc cgacagcagc
     5161 aggaaatgga ggaagaccag cgctggctgg aaaaggaaga gcgattcctg aaacctgatg
     5221 tgaggctctc tcgaggcagc atcgacaggg aggacgggag ttttcagggt ccgactggaa
     5281 accaacacat ctatcagcct gtgggaaaac cagatcctgc agctccacca aagaaaccac
     5341 ctcgccctgg agcacctggc cacctaagca acttgtccag catcagcagc cctgcagaaa
     5401 gctacaatga gggtgtcaag ccgtggaggc ttcagcccca ggaaatcagc ccccctccca
     5461 ctgccaacct tgaccgatcc aatgacaagg tatatgaaaa tgtgacgggc ctagtgaagg
     5521 ctgtcattga gatgtccagc aaaatccaac cagctcctcc agaagagtac gtccctatgg
     5581 tgaaggaagt tggcctggcc ctgcggacct tactggccac tgtggatgag accattccca
     5641 tccttccagc cagcacccat cgagagatcg agatggcaca gaagctgctg aactccgact
     5701 tgggtgagct catcagcaag atgaagctgg cgcagcagta tgtcatgacc agccttcagc
     5761 aggagtacaa gaagcagatg ctgacagctg ctcacgccct ggctgtggat gccaagaatc
     5821 tacttgatgt tattgatcaa gcgagactga aaatgctagg gcagacacgg ccacacgact
     5881 acaaagacca tgacggtgat tataaagatc atgacatcga ctacaaggat gacgatgaca
     5941 agtagaagct taagtttaaa ccgctgatca gcctcgactg tgccttctag ttgccagcca
     6001 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     6061 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     6121 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     6181 ggggatgcgg tgggctctat gggtcgactt cgagcaactt gtttattgca gcttataatg
     6241 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     6301 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc gtctagcatc
     6361 gaagatccaa taacttcgta tagcatacat tatacgaagt tataagtagc ttggcgtaat
     6421 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     6481 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     6541 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     6601 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     6661 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     6721 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     6781 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     6841 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     6901 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     6961 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     7021 aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     7081 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     7141 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     7201 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     7261 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     7321 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     7381 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     7441 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     7501 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     7561 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     7621 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     7681 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     7741 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     7801 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     7861 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     7921 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     7981 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     8041 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     8101 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     8161 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aacacgggat aataccgcgc
     8221 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     8281 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     8341 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     8401 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     8461 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     8521 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     8581 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc actctaggca
     8641 aaatagcacc ctcccgctcc agaacaacat acagcgcttc acagcggcag cctaacagtc
     8701 agccttacca gtaaaaaaga aaacctatta aaaaaacacc actcgacacg gcaccagctc
     8761 aatcagtcac agtgtaaaaa agggccaagt gcagagcgag tatatatagg actaaaaaat
     8821 gacgtaacgg ttaaagtcca caaaaaacac ccagaaaacc gcacgcgaac ctacgcccag
     8881 aaacgaaagc caaaaaaccc acaacttcct caaatcgtca cttccgtttt cccacgttac
     8941 gtaacttccc attttaagaa aactacaatt cccaacacat acaagttact ccgccctaaa
     9001 acctacgtca cccgccccgt tcccacgccc cgcgccacgt cacaaactcc accccctcat
     9061 tatcatattg gcttcaatcc aaaataaggt ata
//
