LOCUS V020151 6923 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V020151 VERSION V020151 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..1603 /label="Cdk5r1(NM_053891)" /note="Cdk5r1(NM_053891)" /gene="Cdk5r1" CDS 1610..1675 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 1699..2287 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 2353..2401 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 2603..3031 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3045..3374 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3441..4235 /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 4411..4532 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4581..4597) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4605..4621 /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(4629..4659) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4674..4695 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4983..5571) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5742..6602) /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(6603..6707) /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 ccatgggcac ggtgctgtcc ctgtccccca gctatcgaaa 721 ggccacactg tttgaggatg gcgcggccac cgtgggccac tacacggccg tgcagaacag 781 caagaacgcc aaggacaaga acctgaagcg gcactccatc atctcggtac tgccttggaa 841 gaggatcgtg gcggtgtcag cgaagaagaa gaactccaag aaggcgcagc ccaacagcag 901 ctaccagagc aacatcgcgc acctcaacaa tgagaacctg aagaagtcgc tgtcgtgtgc 961 caacctgtcc acgttcgccc agcccccgcc ggcccagccg cccgcacccc cagccagcca 1021 gctttctggc tcccagactg gggtctcctc ttctgtcaag aaggccccgc accctgccat 1081 cacctccgca ggaacaccca aacgggtcat cgtccaggcg tccactagtg agctgctgcg 1141 ctgcctgggt gagtttctct gccgccggtg ctaccgcctg aagcacttgt ccccaacgga 1201 ccccgtgctc tggctgcgca gcgtggaccg ctccctgctt ctgcagggat ggcaggacca 1261 gggtttcatc acaccggcca atgtggtctt cctctacatg ctctgcaggg atgttatctc 1321 ctctgaggtg ggctcggacc acgagctcca ggctgtcctg ctgacctgtc tgtacctctc 1381 ctattcctac atgggcaatg aaatctccta cccgctcaag cccttcctgg tggagagctg 1441 caaggaagcc ttttgggacc gttgcctctc agttatcaac ctcatgagct ccaagatgct 1501 gcagatcaat gctgacccac actatttcac acaagtgttc tctgacttga agaatgagag 1561 tggtcaggag gacaagaagc gactcctcct ggggctcgat cggctcgagg actacaaaga 1621 ccatgacggt gattataaag atcatgacat cgactacaag gatgacgatg acaagtagtg 1681 agggcccgat atctcgacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt 1741 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat 1801 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg 1861 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt 1921 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact 1981 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc 2041 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaagctgacg 2101 tcctttccat ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc 2161 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg 2221 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc 2281 tccccgcctg gaaacggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa 2341 cccgcgctat gacggcaata aaaagacaga ataaaacgca cgggtgttgg gtcgtttgtt 2401 cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat 2461 tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa 2521 ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcaga tctgcgcagc 2581 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg 2641 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct 2701 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggc 2761 atccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag 2821 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg 2881 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc 2941 tcggtctatt cttttgattt ataagggatt ttggggattt cggcctattg gttaaaaaat 3001 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt 3061 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt 3121 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc 3181 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc 3241 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg 3301 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc 3361 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga 3421 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg 3481 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg 3541 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt 3601 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg 3661 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat 3721 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat 3781 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg 3841 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg 3901 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc 3961 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc 4021 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg 4081 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg 4141 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca 4201 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgcgaaatg 4261 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat 4321 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg 4381 gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac 4441 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt 4501 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc 4561 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca 4621 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg 4681 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg 4741 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc 4801 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta 4861 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag 4921 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg 4981 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg 5041 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg 5101 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga 5161 agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc 5221 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt 5281 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact 5341 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg 5401 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt 5461 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt 5521 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct 5581 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg 5641 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt 5701 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt 5761 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc 5821 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg 5881 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc 5941 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg 6001 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca 6061 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga 6121 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct 6181 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg 6241 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca 6301 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata 6361 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct 6421 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact 6481 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa 6541 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc 6601 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga 6661 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga 6721 aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg tcgactctca 6781 gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg 6841 aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca 6901 attgcatgaa gaatctgctt agg //