LOCUS V020735 6707 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V020735 VERSION V020735 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..1387 /label="TICAM2(NM_021649)" /note="TICAM2(NM_021649)" /gene="TICAM2" CDS 1394..1459 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 1483..2071 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 2137..2185 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 2387..2815 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2829..3158 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3225..4019 /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 4195..4316 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4365..4381) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4389..4405 /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(4413..4443) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4458..4479 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4767..5355) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5526..6386) /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(6387..6491) /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 ccatgggtat cgggaagtct aaaataaatt cctgccctct 721 ttctctctct tggggtaaaa ggcacagtgt ggatacaagt ccaggatatc atgagtcaga 781 ttccaagaag tctgaagatc tatccttgtg taatgttgct gagcacagca atacaacaga 841 ggggccaaca ggaaagcagg agggagctca gagcgtggaa gagatgtttg aagaagaagc 901 tgaagaagag gtgttcctca aatttgtgat attgcatgca gaagatgaca cagatgaagc 961 cctcagagtc cagaatctgc tacaagatga ctttggtatc aaacccggaa taatctttgc 1021 tgagatgcca tgtggcagac agcatttaca gaatttagat gatgctgtaa atgggtctgc 1081 atggacaatc ttattactga ctgaaaactt tttaagagat acttggtgta atttccagtt 1141 ctatacgtcc ctaatgaact ccgttaacag gcagcataaa tacaactctg ttatacccat 1201 gcggcccctg aacaatcccc ttccccgaga aaggactccc tttgccctcc aaaccatcaa 1261 tgccttagag gaagaaagtc gtggatttcc tacacaagta gaaagaattt ttcaggagtc 1321 tgtgtataag acacaacaaa ctatatggaa agagacaaga aatatggtac aaagacaatt 1381 tattgccctc gaggactaca aagaccatga cggtgattat aaagatcatg acatcgacta 1441 caaggatgac gatgacaagt agtgagggcc cgatatctcg acaatcaacc tctggattac 1501 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga 1561 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc 1621 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa 1681 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc 1741 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc 1801 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc 1861 gtggtgttgt cggggaagct gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg 1921 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct 1981 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg 2041 agtcggatct ccctttgggc cgcctccccg cctggaaacg ggggaggcta actgaaacac 2101 ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa 2161 cgcacgggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct 2221 gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca 2281 ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc 2341 cctgccatag cagatctgcg cagctggggc tctagggggt atccccacgc gccctgtagc 2401 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc 2461 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt 2521 ccccgtcaag ctctaaatcg gggcatccct ttagggttcc gatttagtgc tttacggcac 2581 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag 2641 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa 2701 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgggg 2761 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc 2821 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta 2881 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag 2941 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa 3001 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac 3061 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt 3121 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat 3181 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg 3241 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac 3301 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg 3361 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc 3421 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg 3481 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc 3541 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc 3601 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta 3661 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg 3721 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg 3781 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat 3841 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc 3901 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta 3961 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag 4021 cgggactctg gggttcgcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat 4081 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc 4141 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg 4201 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa 4261 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat 4321 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct 4381 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt 4441 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc 4501 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg 4561 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg 4621 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca 4681 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac 4741 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac 4801 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg 4861 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac 4921 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat 4981 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag 5041 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac 5101 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt 5161 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt 5221 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc 5281 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga 5341 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac 5401 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc 5461 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct 5521 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca 5581 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct 5641 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca 5701 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc 5761 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg 5821 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct 5881 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa 5941 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta 6001 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc 6061 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg 6121 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa 6181 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg 6241 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc 6301 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg 6361 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat 6421 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata 6481 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc 6541 ccgatcccct atggtcgact ctcagtacaa tctgctctga tgccgcatag ttaagccagt 6601 atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta 6661 caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagg //