LOCUS V025969 6431 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V025969 VERSION V025969 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..1921 /label="VSIG2(NM_014312)" /note="VSIG2(NM_014312)" /gene="VSIG2" CDS 1928..1993 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 2031..2255 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2301..2729 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2743..3072 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3139..3933 /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 4107..4228 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4277..4293) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4301..4317 /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(4325..4355) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4370..4391 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4679..5264) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5435..6295) /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(6296..6400) /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 atggccgagc tcccggggcc 961 ctttctctgc ggggccctgc taggcttcct gtgcctgagt gggctggccg tggaggtgaa 1021 ggtacccaca gagccgctga gcacgcccct ggggaagaca gccgagctga cctgcaccta 1081 cagcacgtcg gtgggagaca gcttcgccct ggagtggagc tttgtgcagc ctgggaaacc 1141 catctctgag tcccatccaa tcctgtactt caccaatggc catctgtatc caactggttc 1201 taagtcaaag cgggtcagcc tgcttcagaa cccccccaca gtgggggtgg ccacactgaa 1261 actgactgac gtccacccct cagatactgg aacctacctc tgccaagtca acaacccacc 1321 agatttctac accaatgggt tggggctaat caaccttact gtgctggttc cccccagtaa 1381 tcccttatgc agtcagagtg gacaaacctc tgtgggaggc tctactgcac tgagatgcag 1441 ctcttccgag ggggctccta agccagtgta caactgggtg cgtcttggaa cttttcctac 1501 accttctcct ggcagcatgg ttcaagatga ggtgtctggc cagctcattc tcaccaacct 1561 ctccctgacc tcctcgggca cctaccgctg tgtggccacc aaccagatgg gcagtgcatc 1621 ctgtgagctg accctctctg tgaccgaacc ctcccaaggc cgagtggccg gagctctgat 1681 tggggtgctc ctgggcgtgc tgttgctgtc agttgctgcg ttctgcctgg tcaggttcca 1741 gaaagagagg gggaagaagc ccaaggagac atatgggggt agtgaccttc gggaggatgc 1801 catcgctcct gggatctctg agcacacttg tatgagggct gattctagca aggggttcct 1861 ggaaagaccc tcgtctgcca gcaccgtgac gaccaccaag tccaagctcc ctatggtcgt 1921 gctcgaggac tacaaagacc atgacggtga ttataaagat catgacatcg actacaagga 1981 tgacgatgac aagtagtgag ggcccgttta aacccgctga tcagcctcga ctgtgccttc 2041 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc 2101 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg 2161 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa 2221 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg 2281 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt 2341 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt 2401 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct 2461 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg 2521 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga 2581 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc 2641 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga 2701 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt 2761 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca 2821 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat 2881 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg 2941 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc 3001 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta 3061 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca 3121 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct 3181 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc 3241 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc 3301 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc 3361 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg 3421 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc 3481 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac 3541 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat 3601 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc 3661 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg 3721 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg 3781 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc 3841 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc 3901 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg 3961 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa 4021 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc 4081 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat 4141 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg 4201 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga 4261 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc 4321 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct 4381 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc 4441 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt 4501 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag 4561 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca 4621 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt 4681 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc 4741 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct 4801 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg 4861 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca 4921 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact 4981 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta 5041 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta 5101 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct 5161 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggttttt 5221 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct 5281 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga 5341 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa 5401 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac 5461 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga 5521 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc 5581 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca 5641 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta 5701 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg 5761 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc 5821 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg 5881 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt 5941 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt 6001 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata 6061 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc 6121 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac 6181 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa 6241 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct 6301 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat 6361 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc 6421 cacctgacgt c //