LOCUS       V039939                 6785 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V039939
VERSION     V039939
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..3563
                     /label="Tpm1(NM_001301336)"
                     /note="Tpm1(NM_001301336)"
                     /gene="Tpm1"
     CDS             3570..3635
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3670..3894
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     polyA_signal    3908..4042
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     protein_bind    complement(4062..4095)
                     /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(4117..4133)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4141..4157
                     /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(4165..4195)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4210..4231
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4519..5107)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5278..6138)
                     /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(6139..6243)
                     /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 catggacgcc atcaagaaga agatgcagat gctgaagctc gacaaagaga
     2761 acgccttgga tcgagcagag caggcggagg ctgacaagaa ggctgcggaa gaccggagca
     2821 agcagctgga agatgagctg gtgtcactgc aaaagaaact caagggcact gaagatgaac
     2881 tggacaaata ctccgaggct ctcaaagatg cccaggagaa actggagctg gcggagaaaa
     2941 aggccacaga tgctgaagct gacgtagcat ctctgaacag acgcatccag ctggttgagg
     3001 aggagttgga tcgcgctcag gagcgtctgg ccacagctct acagaagctg gaggaggctg
     3061 agaaggctgc agatgagagt gagagaggca tgaaagtcat tgaaagccga gcccaaaaag
     3121 atgaagaaaa gatggagatt caggagatcc agctgaaaga ggccaagcac attgctgaag
     3181 atgctgaccg aaagtatgaa gaggtggccc gtaagctggt catcatcgag agcgatctgg
     3241 agcgtgcgga ggagagggct gagctctcgg aaggcaaatg tgccgagctt gaagaagagt
     3301 tgaaaacggt gacgaacaac ttgaagtcac tggaggctca ggctgagaag tactctcaga
     3361 aagaagacaa gtatgaagag gagatcaagg ttctctctga caagctgaag gaggctgaga
     3421 cccgggctga gtttgcagag agatcagtaa ccaaattgga gaaaagcatt gatgacttag
     3481 aagacgagct gtatgctcag aaactgaagt acaaggccat cagcgaggag ctggaccacg
     3541 ctctcaacga tatgacttcc ataggatccg actacaaaga ccatgacggt gattataaag
     3601 atcatgacat cgactacaag gatgacgatg acaagtgaag cttaagttta aaccgctgat
     3661 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     3721 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     3781 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     3841 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atgggtcgac
     3901 ttcgagcaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     3961 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     4021 tgtatcttat catgtctgga tcgtctagca tcgaagatcc aataacttcg tatagcatac
     4081 attatacgaa gttataagta gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     4141 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     4201 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     4261 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     4321 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     4381 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     4441 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     4501 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     4561 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     4621 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     4681 ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc
     4741 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     4801 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     4861 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     4921 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     4981 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     5041 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     5101 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     5161 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     5221 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     5281 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     5341 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     5401 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     5461 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     5521 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     5581 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     5641 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     5701 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     5761 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     5821 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     5881 ttgcccggcg tcaacacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     5941 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     6001 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     6061 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     6121 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     6181 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     6241 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     6301 aacctataaa aataggcgta tcactctagg caaaatagca ccctcccgct ccagaacaac
     6361 atacagcgct tcacagcggc agcctaacag tcagccttac cagtaaaaaa gaaaacctat
     6421 taaaaaaaca ccactcgaca cggcaccagc tcaatcagtc acagtgtaaa aaagggccaa
     6481 gtgcagagcg agtatatata ggactaaaaa atgacgtaac ggttaaagtc cacaaaaaac
     6541 acccagaaaa ccgcacgcga acctacgccc agaaacgaaa gccaaaaaac ccacaacttc
     6601 ctcaaatcgt cacttccgtt ttcccacgtt acgtaacttc ccattttaag aaaactacaa
     6661 ttcccaacac atacaagtta ctccgcccta aaacctacgt cacccgcccc gttcccacgc
     6721 cccgcgccac gtcacaaact ccaccccctc attatcatat tggcttcaat ccaaaataag
     6781 gtata
//
