LOCUS V038708 6595 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V038708 VERSION V038708 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..2346 /label="STX4(NM_001272095)" /note="STX4(NM_001272095)" /gene="STX4" CDS 2359..2424 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 2459..2683 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2729..3157 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3171..3500 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3512..4111 /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 4271..4392 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4441..4457) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4465..4481 /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(4489..4519) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4534..4555 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4843..5428) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5599..6459) /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(6460..6564) /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 tgaagcagga gctgcagaac ctgcgcgatg agatcaaaca gctggggagg 1741 gagatccgcc tgcagctgaa ggccatagag ccccagaagg aggaagctga tgagaactat 1801 aactccgtca acacaagaat gagaaaaacc cagcatgggg tcctgtccca gcaattcgtg 1861 gagctcatca acaagtgcaa ttcaatgcag tccgaatacc gggagaagaa cgtggagcgg 1921 attcggaggc agctgaagat caccaatgct gggatggtgt ctgatgagga gttggagcag 1981 atgctggaca gtgggcaaag cgaggtgttt gtgtccaata tcctgaagga cacgcaggtg 2041 actcgacagg ccttaaatga gatctcggcc cggcacagtg agatccagca gcttgaacgc 2101 agtattcgtg agctgcacga catattcact tttctggcta ccgaagtgga gatgcagggg 2161 gagatgatca atcggattga gaagaacatc ctgagctcag cggactacgt ggaacgtggg 2221 caggagcacg tcaagacggc cctggagaac cagaagaagg cgaggaagaa gaaagtcttg 2281 attgccatct gtgtgtccat caccgtcgtc ctcctagcag tcatcattgg cgtcacagtg 2341 gttggactcg agaccggtga ctacaaagac catgacggtg attataaaga tcatgacatc 2401 gattacaagg atgacgatga caagtgaccg gtaagtttaa accgctgatc agcctcgact 2461 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg 2521 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg 2581 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg 2641 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga 2701 accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg 2761 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct 2821 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat 2881 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt 2941 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg 3001 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac 3061 cctatctcgg tctattcttt tgatttataa gggattttgg ccatttcggc ctattggtta 3121 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt 3181 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca 3241 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa 3301 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc 3361 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg 3421 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg 3481 gaggcctagg cttttgcaaa aagctcccgg gatgaccgag tacaagccca cggtgcgcct 3541 cgccacccgc gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta 3601 ccccgccacg cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca 3661 agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg 3721 cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga 3781 gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga 3841 aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt 3901 ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc 3961 ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt 4021 ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac 4081 ctggtgcatg acccgcaagc ccggtgcctg attcgaaatg accgaccaag cgacgcccaa 4141 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat 4201 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt 4261 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac 4321 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat 4381 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg 4441 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc 4501 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc 4561 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat 4621 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac 4681 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt 4741 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca 4801 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc 4861 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact 4921 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct 4981 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag 5041 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca 5101 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa 5161 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc 5221 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag 5281 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg 5341 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ttttttgttt gcaagcagca 5401 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga 5461 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat 5521 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga 5581 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg 5641 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga 5701 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc 5761 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac 5821 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc 5881 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc 5941 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc 6001 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt 6061 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc 6121 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg 6181 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag 6241 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat 6301 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc 6361 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa 6421 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta 6481 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa 6541 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtc //