LOCUS V018344 7127 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V018344 VERSION V018344 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers enhancer 50..429 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 430..633 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 683..1774 /label="HAVCR1(NM_012206)" /note="HAVCR1(NM_012206)" /gene="HAVCR1" CDS 1814..1879 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 1903..2491 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 2557..2605 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 2807..3235 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3249..3578 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3645..4439 /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 4615..4736 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4785..4801) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4809..4825 /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(4833..4863) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4878..4899 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5187..5775) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5946..6806) /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(6807..6911) /gene="bla" /label="AmpR promoter" ORIGIN 1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta 61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag 121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc 181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga 241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat 301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc 361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct 421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca 481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat 541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg 601 cgtgtacggt gggaggtcta tataagcaga gctctggcta gcgtttaaac ttaagcttgg 661 taccgagctc ggatccgcca ccatgcatcc tcaagtggtc atcttaagcc tcatcctaca 721 tctggcagat tctgtagctg gttctgtaaa ggttggtgga gaggcaggtc catctgtcac 781 actaccctgc cactacagtg gagctgtcac atccatgtgc tggaatagag gctcatgttc 841 tctattcaca tgccaaaatg gcattgtctg gaccaatgga acccacgtca cctatcggaa 901 ggacacacgc tataagctat tgggggacct ttcaagaagg gatgtctctt tgaccataga 961 aaatacagct gtgtctgaca gtggcgtata ttgttgccgt gttgagcacc gtgggtggtt 1021 caatgacatg aaaatcaccg tatcattgga gattgtgcca cccaaggtca cgactactcc 1081 aattgtcaca actgttccaa ccgtcacgac tgttcgaacg agcaccactg ttccaacgac 1141 aacgactgtt ccaatgacga ctgttccaac gacaactgtt ccaacaacaa tgagcattcc 1201 aacgacaacg actgttctga cgacaatgac tgtttcaacg acaacgagcg ttccaacgac 1261 aacgagcatt ccaacaacaa caagtgttcc agtgacaaca actgtctcta cctttgttcc 1321 tccaatgcct ttgcccaggc agaaccatga accagtagcc acttcaccat cttcacctca 1381 gccagcagaa acccacccta cgacactgca gggagcaata aggagagaac ccaccagctc 1441 accattgtac tcttacacaa cagatgggaa tgacaccgtg acagagtctt cagatggcct 1501 ttggaataac aatcaaactc aactgttcct agaacatagt ctactgacgg ccaataccac 1561 taaaggaatc tatgctggag tctgtatttc tgtcttggtg cttcttgctc ttttgggtgt 1621 catcattgcc aaaaagtatt tcttcaaaaa ggaggttcaa caactaagtg tttcatttag 1681 cagccttcaa attaaagctt tgcaaaatgc agttgaaaag gaagtccaag cagaagacaa 1741 tatctacatt gagaatagtc tttatgccac ggacgaattc tgcagatatc cagcacagtg 1801 gcggccgctc gaggactaca aagaccatga cggtgattat aaagatcatg acatcgacta 1861 caaggatgac gatgacaagt agtgagggcc cgatatctcg acaatcaacc tctggattac 1921 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga 1981 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc 2041 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa 2101 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc 2161 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc 2221 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc 2281 gtggtgttgt cggggaagct gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg 2341 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct 2401 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg 2461 agtcggatct ccctttgggc cgcctccccg cctggaaacg ggggaggcta actgaaacac 2521 ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa 2581 cgcacgggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct 2641 gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca 2701 ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc 2761 cctgccatag cagatctgcg cagctggggc tctagggggt atccccacgc gccctgtagc 2821 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc 2881 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt 2941 ccccgtcaag ctctaaatcg gggcatccct ttagggttcc gatttagtgc tttacggcac 3001 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag 3061 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa 3121 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgggg 3181 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc 3241 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta 3301 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag 3361 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa 3421 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac 3481 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt 3541 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat 3601 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg 3661 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac 3721 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg 3781 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc 3841 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg 3901 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc 3961 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc 4021 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta 4081 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg 4141 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg 4201 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat 4261 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc 4321 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta 4381 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag 4441 cgggactctg gggttcgcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat 4501 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc 4561 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg 4621 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa 4681 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat 4741 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct 4801 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt 4861 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc 4921 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg 4981 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg 5041 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca 5101 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac 5161 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac 5221 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg 5281 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac 5341 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat 5401 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag 5461 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac 5521 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt 5581 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt 5641 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc 5701 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga 5761 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac 5821 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc 5881 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct 5941 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca 6001 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct 6061 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca 6121 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc 6181 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg 6241 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct 6301 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa 6361 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta 6421 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc 6481 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg 6541 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa 6601 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg 6661 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc 6721 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg 6781 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat 6841 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata 6901 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc 6961 ccgatcccct atggtcgact ctcagtacaa tctgctctga tgccgcatag ttaagccagt 7021 atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta 7081 caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagg //