LOCUS V020095 7484 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V020095 VERSION V020095 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..2164 /label="ITGBL1(NM_004791)" /note="ITGBL1(NM_004791)" /gene="ITGBL1" CDS 2171..2236 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 2260..2848 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 2914..2962 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 3164..3592 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3606..3935 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4002..4796 /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 4972..5093 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5142..5158) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5166..5182 /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(5190..5220) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5235..5256 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5544..6132) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6303..7163) /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(7164..7268) /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 ccatgcgtcc cccaggcttc aggaacttct tgctgctggc 721 gtcctccctt ctctttgctg ggttgtcagc tgttcctcaa agcttctcgc catctctgag 781 gagctggccg ggcgccgcct gcaggctgtc ccgggccgag tcggagcgac gctgccgcgc 841 acctgggcag cccccggggg ccgcgctgtg ccacggccgg ggccgctgcg actgcggcgt 901 ctgcatctgc cacgtgactg agccgggcat gttcttcggg cccctgtgtg agtgccatga 961 gtgggtgtgc gagacctacg acgggagcac ctgtgcaggc catggtaagt gtgactgtgg 1021 caagtgcaag tgtgaccagg gatggtatgg ggatgcttgc cagtacccaa ctaactgtga 1081 cttgacaaag aagaaaagta accaaatgtg caagaattca caagacatca tctgctctaa 1141 tgcaggtaca tgtcactgtg gcaggtgtaa gtgtgataat tcagatggaa gtggacttgt 1201 gtatggtaaa ttttgtgagt gtgacgatag agaatgcata gacgatgaaa cagaagaaat 1261 atgtggaggc catgggaagt gttactgtgg aaactgctac tgcaaggctg gttggcatgg 1321 agataaatgt gaattccagt gcgatatcac cccctgggaa agcaagcgaa gatgcacgtc 1381 tccagatggc aaaatctgca gtaacagagg gacttgtgta tgtggtgaat gtacctgtca 1441 cgatgttgat ccgactgggg actggggaga tattcatggg gacacctgtg aatgtgatga 1501 gagggactgt agagctgtct atgaccgata ttctgatgac ttctgttcag gtcatggaca 1561 gtgtaattgc ggaagatgtg actgcaaagc aggctggtat gggaagaagt gtgagcaccc 1621 acagtcctgc acgctgtcag ctgaggagag catcaggaag tgccagggaa gctcggatct 1681 gccttgctct gggaggggta aatgtgaatg tggcaaatgc acctgctatc ctccaggaga 1741 tcgccgggtg tatggcaaga cttgtgagtg tgatgatcgc cgctgtgaag acctcgatgg 1801 tgtggtctgt ggaggccacg gcacatgttc ctgtggtcgc tgtgtttgtg agagaggatg 1861 gtttggaaag ctctgccaac atccgcggaa gtgtaacatg acggaagaac aaagcaagaa 1921 tctgtgtgaa tcagcagatg gcatattgtg ctcggggaag ggttcttgtc attgtgggaa 1981 gtgcatttgt tctgctgaag agtggtatat ttctggggag ttctgtgact gtgatgacag 2041 agactgcgac aaacatgatg gtctcatttg tacagggaat ggaatatgta gctgtggaaa 2101 ctgtgaatgc tgggatggat ggaatggaaa tgcatgtgaa atctggcttg gctcagaata 2161 tcctctcgag gactacaaag accatgacgg tgattataaa gatcatgaca tcgactacaa 2221 ggatgacgat gacaagtagt gagggcccga tatctcgaca atcaacctct ggattacaaa 2281 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac 2341 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc 2401 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt 2461 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc 2521 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc 2581 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg 2641 gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt 2701 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc 2761 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt 2821 cggatctccc tttgggccgc ctccccgcct ggaaacgggg gaggctaact gaaacacgga 2881 aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc 2941 acgggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc 3001 gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc 3061 caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct 3121 gccatagcag atctgcgcag ctggggctct agggggtatc cccacgcgcc ctgtagcggc 3181 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc 3241 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc 3301 cgtcaagctc taaatcgggg catcccttta gggttccgat ttagtgcttt acggcacctc 3361 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg 3421 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact 3481 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttggggatt 3541 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt 3601 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc 3661 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag 3721 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc 3781 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa 3841 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt 3901 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca 3961 ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat 4021 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac 4081 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc 4141 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc 4201 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag 4261 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc 4321 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg 4381 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc 4441 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc 4501 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga 4561 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca 4621 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg 4681 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg 4741 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg 4801 gactctgggg ttcgcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc 4861 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc 4921 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt 4981 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca 5041 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc 5101 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg 5161 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa 5221 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct 5281 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga 5341 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc 5401 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa 5461 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt 5521 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa 5581 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt 5641 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg 5701 tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc 5761 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc 5821 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta 5881 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct 5941 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc 6001 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa 6061 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa 6121 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa 6181 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt 6241 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac 6301 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc 6361 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc 6421 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata 6481 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc 6541 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc 6601 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca 6661 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa 6721 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca 6781 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt 6841 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt 6901 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg 6961 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga 7021 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc 7081 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg 7141 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag 7201 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg 7261 gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg 7321 atcccctatg gtcgactctc agtacaatct gctctgatgc cgcatagtta agccagtatc 7381 tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa 7441 caaggcaagg cttgaccgac aattgcatga agaatctgct tagg //