LOCUS V040543 6805 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V040543 VERSION V040543 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 928..2241 /label="VIPR2(NM_003382)" /note="VIPR2(NM_003382)" /gene="VIPR2" CDS 2248..2271 /codon_start=1 /product="FLAG® epitope tag, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDDDDK" /label="FLAG" CDS 2278..2367 /codon_start=1 /product="two Strep-Tag®II moieties connected by a linker for enhanced binding to Strep-Tactin®, an engineered form of streptavidin (Schmidt et al., 2013)" /transl_table=1 /translation="SA,WSHPQFEK,GGGSGGGSGGSA,WSHPQFEK" /label="Twin-Strep-tag" polyA_signal 2405..2629 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2675..3103 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3117..3446 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3513..4307 /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 4481..4602 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4651..4667) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4675..4691 /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(4699..4729) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4744..4765 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5053..5638) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5809..6669) /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(6670..6774) /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 aagctgaatt cgccaccatg cggacgctgc tgcctcccgc gctgctgacc 961 tgctggctgc tcgcccccgt gaacagcatt cacccagaat gccgatttca tctggaaata 1021 caggaggaag aaacaaaatg tgcagagctt ctgaggtctc aaacagaaaa acacaaagcc 1081 tgcagtggcg tctgggacaa catcacgtgc tggcggcctg ccaatgtggg agagaccgtc 1141 acggtgccct gcccaaaagt cttcagcaat ttttacagca aagcaggaaa cataagcaaa 1201 aactgtacga gtgacggatg gtcagagacg ttcccagatt tcgtcgatgc ctgtggctac 1261 agcgacccgg aggatgagag caagatcacg ttttatattc tggtgaaggc catttataca 1321 ctgggctaca gtgtctctct gatgtctctt gcaacaggaa gcataattct gtgcctcttc 1381 aggaagctgc actgcaccag gaattacatc cacctgaacc tgttcctgtc cttcatcctg 1441 agagccatct cagtgctggt caaggacgac gttctctact ccagctctgg cacgttgcac 1501 tgccctgacc agccatcctc ctgggtgggc tgcaagctga gcctggtctt cctgcagtac 1561 tgcatcatgg ccaacttctt ctggctgctg gtggaggggc tctacctcca caccctcctg 1621 gtggccatgc tcccccctag aaggtgcttc ctggcctacc tcctgatcgg atggggcctc 1681 cccaccgtct gcatcggtgc atggactgcg gccaggctct acttagaaga caccggttgc 1741 tgggatacaa acgaccacag tgtgccctgg tgggtcatac gaataccgat tttaatttcc 1801 atcatcgtca attttgtcct tttcattagt attatacgaa ttttgctgca gaagttaaca 1861 tccccagatg tcggcggcaa cgaccagtct cagtacaaga ggctggccaa gtccacgctc 1921 ctgcttatcc cgctgttcgg cgtccactac atggtgtttg ccgtgtttcc catcagcatc 1981 tcctccaaat accagatact gtttgagctg tgcctcgggt cgttccaggg cctggtggtg 2041 gccgtcctct actgtttcct gaacagtgag gtgcagtgcg agctgaagcg aaaatggcga 2101 agccggtgcc cgaccccgtc cgcgagccgg gattacaggg tctgcggttc ctccttctcc 2161 cgcaacggct cggagggcgc cctgcagttc caccgcggct cccgcgccca gtccttcctg 2221 caaacggaga cctcggtcat cctcgaggat tacaaggatg acgacgataa gggatccagc 2281 gcctggtccc accctcagtt tgagaagggc ggaggctctg gcggcggaag cggaggatct 2341 gcttggagcc acccccagtt cgaaaagtga tgagggcccg tttaaacccg ctgatcagcc 2401 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg 2461 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat 2521 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag 2581 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg 2641 gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc 2701 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc 2761 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct 2821 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa 2881 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc 2941 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca 3001 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat 3061 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt 3121 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc 3181 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta 3241 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc 3301 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta 3361 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct 3421 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat 3481 ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag 3541 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg 3601 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca 3661 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc 3721 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg 3781 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg 3841 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta 3901 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag 3961 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac 4021 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg 4081 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg 4141 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg 4201 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg 4261 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg 4321 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc 4381 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct 4441 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta 4501 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact 4561 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc 4621 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta 4681 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc 4741 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg 4801 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg 4861 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg 4921 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa 4981 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc 5041 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc 5101 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag 5161 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct 5221 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta 5281 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc 5341 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc 5401 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt 5461 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct 5521 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc 5581 tggtagcggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga 5641 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg 5701 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg 5761 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt 5821 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact 5881 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat 5941 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg 6001 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg 6061 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat 6121 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc 6181 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt 6241 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc 6301 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga 6361 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc 6421 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa 6481 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta 6541 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg 6601 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg 6661 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat 6721 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt 6781 tccccgaaaa gtgccacctg acgtc //