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 //