LOCUS V022887 6515 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V022887 VERSION V022887 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 941..2005 /label="VASH2(NM_001301056)" /note="VASH2(NM_001301056)" /gene="VASH2" CDS 2012..2077 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 2115..2339 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2385..2813 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2827..3156 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3223..4017 /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 4191..4312 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4361..4377) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4385..4401 /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(4409..4439) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4454..4475 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4763..5348) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5519..6379) /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(6380..6484) /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 aagcttggta ccgagctcgg atccgccacc atgaccggct ccgcggccga 961 cactcaccgc tgcccccacc ccaaaggcgc caaaggcacc cggtcccgga gcagccacgc 1021 gcggcccgtg agcctcgcca ccagcggggg ctcagaggag gaggacaaag acggcggggt 1081 gctgttccac gtcaacaaga gcggcttccc catcgacagc cacacctggg agcgcatgtg 1141 gatgcacgtg gccaaggtgc accctaaggg gggagaaatg gtgggcgcca tcaggaacgc 1201 cgccttcttg gcaaagcctt caatacccca ggtcccaaac tacaggctgt cgatgacgat 1261 cccagactgg ctccaggcga tccagaatta catgaagacc ctacaatata atcacacagg 1321 gacccagttc tttgaaatta ggaaaatgag accgctgagt gggttaatgg aaacagcaaa 1381 agaaatgacc cgagagtcct tgcctatcaa atgccttgaa gctgtcatcc tgggcatcta 1441 cttaaccaat gggcagcctt ccattgagcg gttccccatc agctttaaaa cctacttctc 1501 aggaaactac tttcaccacg ttgtgctggg gatttactgc aatggccgct atggctcatt 1561 gggcatgagc cgcagggctg agctgatgga caagccattg acttttcgga ctctgagtga 1621 cctcatcttt gactttgagg actcttacaa gaaatacctg cacacagtca agaaggtcaa 1681 gattgggctg tacgtccccc atgagcctca tagcttccag cccattgagt ggaagcagct 1741 ggtcctcaac gtctcaaaga tgctgagggc tgacataagg aaggagctgg agaaatatgc 1801 cagggacatg agaatgaaga tcctgaaacc tgcaagtgcc cactctccga cccaagtgag 1861 aagccgggga aaatccctgt cccccagaag gagacaggca agccccccga ggaggctcgg 1921 ccggcgagag aagtcgcctg cactgcctga aaagaaggtg gctgatctga gcactctgaa 1981 tgaagtgggc tatcaaatcc gaattctcga ggactacaaa gaccatgacg gtgattataa 2041 agatcatgac atcgactaca aggatgacga tgacaagtag tgagggcccg tttaaacccg 2101 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt 2161 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat 2221 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag 2281 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc 2341 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg 2401 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc 2461 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc 2521 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct 2581 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac 2641 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac 2701 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat 2761 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg 2821 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg 2881 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca 2941 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact 3001 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta 3061 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag 3121 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc 3181 attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga 3241 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa 3301 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt 3361 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg 3421 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa 3481 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac 3541 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt 3601 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact 3661 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg 3721 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg 3781 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc 3841 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt 3901 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc 3961 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg 4021 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg 4081 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct 4141 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta 4201 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat 4261 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct 4321 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt 4381 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag 4441 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt 4501 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag 4561 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg 4621 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat 4681 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta 4741 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa 4801 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc 4861 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt 4921 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca 4981 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg 5041 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat 5101 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta 5161 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct 5221 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac 5281 aaaccaccgc tggtagcggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag 5341 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact 5401 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa 5461 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt 5521 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag 5581 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca 5641 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc 5701 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt 5761 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg 5821 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca 5881 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg 5941 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca 6001 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg 6061 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct 6121 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca 6181 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca 6241 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg 6301 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac 6361 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt 6421 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc 6481 cgcgcacatt tccccgaaaa gtgccacctg acgtc //