LOCUS V019223 7043 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V019223 VERSION V019223 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..1723 /label="JUND(NM_005354)" /note="JUND(NM_005354)" /gene="JUND" CDS 1730..1795 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 1819..2407 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 2473..2521 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 2723..3151 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3165..3494 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3561..4355 /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 4531..4652 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4701..4717) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4725..4741 /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(4749..4779) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4794..4815 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5103..5691) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5862..6722) /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(6723..6827) /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 ccatggaaac acccttctac ggcgatgagg cgctgagcgg 721 cctgggcggc ggcgccagtg gcagcggcgg cagcttcgcg tccccgggcc gcttgttccc 781 cggggcgccc ccgacggccg cggccggcag catgatgaag aaggacgcgc tgacgctgag 841 cctgagtgag caggtggcgg cagcgctcaa gcctgcggcc gcgccgcctc ctacccccct 901 gcgcgccgac ggcgccccca gcgcggcacc ccccgacggc ctgctcgcct ctcccgacct 961 ggggctgctg aagctggcct cccccgagct cgagcgcctc atcatccagt ccaacgggct 1021 ggtcaccacc acgccgacga gctcacagtt cctctacccc aaggtggcgg ccagcgagga 1081 gcaggagttc gccgagggct tcgtcaaggc cctggaggat ttacacaagc agaaccagct 1141 cggcgcgggc gcggccgctg ccgccgccgc cgccgccgcc ggggggccct cgggcacggc 1201 cacgggctcc gcgccccccg gcgagctggc cccggcggcg gccgcgcccg aagcgcctgt 1261 ctacgcgaac ctgagcagct acgcgggcgg cgccgggggc gcggggggcg ccgcgacggt 1321 cgccttcgct gccgaacctg tgcccttccc gccgccgcca cccccaggcg cgttggggcc 1381 gccgcgcctg gctgcgctca aggacgagcc acagacggtg cccgacgtgc cgagcttcgg 1441 cgagagcccg ccgttgtcgc ccatcgacat ggacacgcag gagcgcatca aggcggagcg 1501 caagcggctg cgcaaccgca tcgccgcctc caagtgccgc aagcgcaagc tggagcgcat 1561 ctcgcgcctg gaagagaaag tgaagaccct caagagtcag aacacggagc tggcgtccac 1621 ggcgagcctg ctgcgcgagc aggtggcgca gctcaagcag aaagtcctca gccacgtcaa 1681 cagcggctgc cagctgctgc cccagcacca ggtgcccgcg tacctcgagg actacaaaga 1741 ccatgacggt gattataaag atcatgacat cgactacaag gatgacgatg acaagtagtg 1801 agggcccgat atctcgacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt 1861 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat 1921 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg 1981 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt 2041 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact 2101 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc 2161 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaagctgacg 2221 tcctttccat ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc 2281 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg 2341 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc 2401 tccccgcctg gaaacggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa 2461 cccgcgctat gacggcaata aaaagacaga ataaaacgca cgggtgttgg gtcgtttgtt 2521 cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat 2581 tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa 2641 ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcaga tctgcgcagc 2701 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg 2761 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct 2821 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggc 2881 atccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag 2941 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg 3001 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc 3061 tcggtctatt cttttgattt ataagggatt ttggggattt cggcctattg gttaaaaaat 3121 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt 3181 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt 3241 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc 3301 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc 3361 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg 3421 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc 3481 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga 3541 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg 3601 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg 3661 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt 3721 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg 3781 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat 3841 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat 3901 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg 3961 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg 4021 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc 4081 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc 4141 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg 4201 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg 4261 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca 4321 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgcgaaatg 4381 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat 4441 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg 4501 gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac 4561 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt 4621 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc 4681 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca 4741 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg 4801 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg 4861 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc 4921 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta 4981 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag 5041 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg 5101 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg 5161 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg 5221 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga 5281 agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc 5341 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt 5401 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact 5461 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg 5521 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt 5581 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt 5641 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct 5701 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg 5761 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt 5821 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt 5881 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc 5941 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg 6001 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc 6061 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg 6121 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca 6181 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga 6241 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct 6301 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg 6361 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca 6421 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata 6481 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct 6541 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact 6601 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa 6661 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc 6721 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga 6781 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga 6841 aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg tcgactctca 6901 gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg 6961 aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca 7021 attgcatgaa gaatctgctt agg //