LOCUS V040995 6907 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V040995 VERSION V040995 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 937..1653 /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 1690..2658 /label="Ppp1cc(NM_013636)" /note="Ppp1cc(NM_013636)" /gene="Ppp1cc" CDS 2671..2736 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 2771..2995 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 3041..3469 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3483..3812 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3824..4423 /codon_start=1 /gene="pac from Streptomyces alboniger" /note="confers resistance to puromycin" /product="puromycin N-acetyltransferase" /transl_table=1 /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*" /label="PuroR" polyA_signal 4583..4704 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4753..4769) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4777..4793 /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(4801..4831) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4846..4867 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5155..5740) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5911..6771) /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(6772..6876) /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 aagctttcta gagaattcga tatcggatcc gccaccatgg tgagcaaggg cgaggagctg 961 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc 1021 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc 1081 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc 1141 gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc 1201 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag 1261 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc 1321 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc 1381 cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc 1441 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc 1501 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg 1561 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc 1621 gggatcactc tcggcatgga cgagctgtac aagggcggtg gaggcagtgg tggaggcggt 1681 agcggtacca tggcggatat cgacaaactc aacatcgaca gcatcatcca acggctgctg 1741 gaagtgagag ggtccaagcc aggcaagaat gtccagctcc aggagaacga gatccgagga 1801 ctctgcctga agtctcggga gatcttcctc agtcagccta tccttttaga acttgaagca 1861 ccactcaaga tatgtggtga catccacggg cagtactatg atttgctccg tctgtttgaa 1921 tacggtggct ttcctccaga gagcaactat ttgtttctcg gggactatgt ggacaggggc 1981 aagcagtccc tggagacaat ctgcctcttg ctggcctaca aaatcaagta tccggagaac 2041 ttctttcttc tcagagggaa ccacgagtgc gccagcatca ataggatcta cggattttat 2101 gatgagtgta aaagaagata caacattaag ctgtggaaaa cgttcacaga ctgttttaac 2161 tgcttgccga tagcagccat cgtggacgag aagatattct gctgtcatgg aggtttatca 2221 ccagatcttc aatctatgga gcagattcgg cgaattatga gaccaactga tgtaccagat 2281 caaggtcttc tttgtgatct tttgtggtct gaccccgata aagatgtctt aggctggggt 2341 gaaaatgaca gaggagtgtc cttcacattt ggtgcagaag tggttgcaaa atttctccat 2401 aagcatgatt tggatcttat atgtagagcc catcaggtgg ttgaagatgg ctatgagttt 2461 tttgcaaaga ggcagttagt cactctgttt tctgcaccca actactgtgg cgagtttgac 2521 aatgcaggcg ccatgatgag tgtggatgag accctcatgt gttccttcca gattttaaag 2581 cctgcagaga aaaagaagcc caacgccacg agacctgtca caccaccacg gggtatgatc 2641 acaaagcaag caaagaaact cgagaccggt gactacaaag accatgacgg tgattataaa 2701 gatcatgaca tcgattacaa ggatgacgat gacaagtgac cggtaagttt aaaccgctga 2761 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct 2821 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca 2881 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag 2941 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct 3001 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca 3061 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta 3121 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt 3181 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac 3241 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt 3301 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga 3361 acaacactca accctatctc ggtctattct tttgatttat aagggatttt ggccatttcg 3421 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga 3481 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa 3541 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca 3601 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc 3661 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt 3721 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag 3781 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggatgaccg agtacaagcc 3841 cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc 3901 gttcgccgac taccccgcca cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt 3961 caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt 4021 cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc 4081 ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca 4141 gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc 4201 caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc 4261 cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg 4321 caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga 4381 aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgattcgaaa tgaccgacca 4441 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt 4501 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat 4561 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag 4621 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt 4681 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt 4741 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca 4801 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact 4861 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct 4921 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc 4981 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca 5041 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg 5101 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca 5161 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa 5221 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc 5281 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc 5341 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct 5401 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg 5461 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag 5521 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta 5581 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg 5641 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtttttttgt 5701 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc 5761 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt 5821 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta 5881 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat 5941 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac 6001 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg 6061 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag 6121 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt 6181 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt 6241 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt 6301 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt 6361 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct 6421 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt 6481 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac 6541 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa 6601 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa 6661 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca 6721 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct 6781 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga 6841 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc 6901 tgacgtc //