LOCUS V047602 6845 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V047602 VERSION V047602 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 923..1639 /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 1676..2335 /label="VAMP7(NM_005638)" /note="VAMP7(NM_005638)" /gene="VAMP7" CDS 2342..2407 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 2445..2669 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2715..3143 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3157..3486 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3553..4347 /codon_start=1 /gene="aph(3')-II (or nptII)" /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin®)" /product="aminoglycoside phosphotransferase from Tn5" /transl_table=1 /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*" /label="NeoR/KanR" polyA_signal 4521..4642 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4691..4707) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4715..4731 /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(4739..4769) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4784..4805 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5093..5678) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5849..6709) /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(6710..6814) /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 gtttaaactt aagcttgcca ccatggtgag caagggcgag gagctgttca ccggggtggt 961 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga 1021 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa 1081 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag 1141 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta 1201 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt 1261 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga 1321 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat 1381 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga 1441 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc 1501 cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa 1561 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg 1621 catggacgag ctgtacaagg gcggtggagg cagtggtgga ggcggtagcg aattcatggc 1681 gattcttttt gctgttgttg ccagggggac cactatcctt gccaaacatg cttggtgtgg 1741 aggaaacttc ctggaggtga cagagcagat tctggctaag ataccttctg aaaataacaa 1801 actaacgtac tcacatggca attatttgtt tcattacatc tgccaagaca ggattgtata 1861 tctttgtatc actgatgatg attttgaacg ttcccgagcc tttaattttc tgaatgagat 1921 aaagaagagg ttccagacta cttacggttc aagagcacag acagcacttc catatgccat 1981 gaatagcgag ttctcaagtg tcttagctgc acagctgaag catcactctg agaataaggg 2041 cctagacaaa gtgatggaga ctcaagccca agtggatgaa ctgaaaggaa tcatggtcag 2101 aaacatagat ctggtagctc agcgaggaga aagattggaa ttattgattg acaaaacaga 2161 aaatcttgtg gattcttctg tcaccttcaa aactaccagc agaaatcttg ctcgagccat 2221 gtgtatgaag aacctcaagc tcactattat catcatcatc gtatcaattg tgttcatcta 2281 tatcattgtt tcacctctct gtggtggatt tacatggcca agctgtgtga agaaactcga 2341 ggactacaaa gaccatgacg gtgattataa agatcatgac atcgactaca aggatgacga 2401 tgacaagtag tgagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg 2461 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc 2521 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc 2581 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag 2641 gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc 2701 tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac 2761 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc 2821 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt 2881 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg 2941 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac 3001 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta 3061 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat 3121 ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag 3181 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc 3241 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat 3301 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt 3361 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc 3421 gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt 3481 tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga 3541 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg 3601 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg 3661 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc 3721 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct 3781 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa 3841 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg 3901 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa 3961 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat 4021 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg 4081 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc 4141 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac 4201 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg 4261 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc 4321 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag 4381 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg 4441 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc 4501 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca 4561 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt 4621 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg 4681 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca 4741 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca 4801 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc 4861 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt 4921 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact 4981 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag 5041 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata 5101 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc 5161 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg 5221 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc 5281 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg 5341 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc 5401 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga 5461 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg 5521 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa 5581 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ttttttgttt 5641 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta 5701 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat 5761 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa 5821 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct 5881 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta 5941 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct 6001 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg 6061 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa 6121 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt 6181 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta 6241 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca 6301 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta 6361 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct 6421 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg 6481 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac 6541 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact 6601 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa 6661 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt 6721 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat 6781 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg 6841 acgtc //