LOCUS V019782 6902 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V019782 VERSION V019782 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..1558 /label="CDK5(NM_004935)" /note="CDK5(NM_004935)" /gene="CDK5" CDS 1565..1654 /codon_start=1 /product="3 tandem Myc epitope tags" /transl_table=1 /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL" /label="3xMyc" misc_feature 1678..2266 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 2332..2380 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 2582..3010 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3024..3353 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3420..4214 /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 4390..4511 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4560..4576) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4584..4600 /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(4608..4638) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4653..4674 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4962..5550) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5721..6581) /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(6582..6686) /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 ccatgcagaa atacgagaaa ctggaaaaga ttggggaagg 721 cacctacgga actgtgttca aggccaaaaa ccgggagact catgagatcg tggctctgaa 781 acgggtgagg ctggatgacg atgatgaggg tgtgccgagt tccgccctcc gggagatctg 841 cctactcaag gagctgaagc acaagaacat cgtcaggctt catgacgtcc tgcacagcga 901 caagaagctg actttggttt ttgaattctg tgaccaggac ctgaagaagt attttgacag 961 ttgcaatggt gacctcgatc ctgagattgt aaagtcattc ctcttccagc tactaaaagg 1021 gctgggattc tgtcatagcc gcaatgtgct acacagggac ctgaagcccc agaacctgct 1081 aataaacagg aatggggagc tgaaattggc tgattttggc ctggctcgag cctttgggat 1141 tcccgtccgc tgttactcag ctgaggtggt cacactgtgg taccgcccac cggatgtcct 1201 ctttggggcc aagctgtact ccacgtccat cgacatgtgg tcagccggct gcatctttgc 1261 agagctggcc aatgctgggc ggcctctttt tcccggcaat gatgtcgatg accagttgaa 1321 gaggatcttc cgactgctgg ggacgcccac cgaggagcag tggccctcta tgaccaagct 1381 gccagactat aagccctatc cgatgtaccc ggccacaaca tccctggtga acgtcgtgcc 1441 caaactcaat gccacaggga gggatctgct gcagaacctt ctgaagtgta accctgtcca 1501 gcgtatctca gcagaagagg ccctgcagca cccctacttc tccgacttct gtccgcccct 1561 cgaggaacaa aaactaatat cagaggaaga tttggaacag aaactaatca gtgaagaaga 1621 tttagagcaa aagttaattt ccgaggagga cttatagtga gggcccgata tctcgacaat 1681 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct 1741 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg 1801 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg 1861 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt 1921 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt 1981 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg 2041 ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg gctgctcgcc 2101 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat 2161 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc 2221 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg aaacggggga 2281 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa 2341 aaagacagaa taaaacgcac gggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc 2401 cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt 2461 tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac 2521 gtcggggcgg caggccctgc catagcagat ctgcgcagct ggggctctag ggggtatccc 2581 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc 2641 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc 2701 acgttcgccg gctttccccg tcaagctcta aatcggggca tccctttagg gttccgattt 2761 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg 2821 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt 2881 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta 2941 taagggattt tggggatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt 3001 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc 3061 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt 3121 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca 3181 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc 3241 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg 3301 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc 3361 cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca 3421 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg 3481 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag 3541 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc 3601 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc 3661 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg 3721 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc 3781 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca 3841 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag 3901 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg 3961 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg 4021 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca 4081 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc 4141 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg 4201 acgagttctt ctgagcggga ctctggggtt cgcgaaatga ccgaccaagc gacgcccaac 4261 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc 4321 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc 4381 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca 4441 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc 4501 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg 4561 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc 4621 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg 4681 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc 4741 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact 4801 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta 4861 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag 4921 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc 4981 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta 5041 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg 5101 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc 5161 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac 5221 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac 5281 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg 5341 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga 5401 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt 5461 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag 5521 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct 5581 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg 5641 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat 5701 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc 5761 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg 5821 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct 5881 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca 5941 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg 6001 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg 6061 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc 6121 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag 6181 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg 6241 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag 6301 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat 6361 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg 6421 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca 6481 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca 6541 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat 6601 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag 6661 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac 6721 ggatcgggag atctcccgat cccctatggt cgactctcag tacaatctgc tctgatgccg 6781 catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga 6841 gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta 6901 gg //