LOCUS V029217 7432 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V029217 VERSION V029217 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 706..2088 /label="CALCRL(NM_005795)" /note="CALCRL(NM_005795)" /gene="CALCRL" CDS 2095..2184 /codon_start=1 /product="3 tandem Myc epitope tags" /transl_table=1 /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL" /label="3xMyc" misc_feature 2208..2796 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 2862..2910 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 3112..3540 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3554..3883 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3950..4744 /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 4920..5041 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5090..5106) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5114..5130 /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(5138..5168) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5183..5204 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5492..6080) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6251..7111) /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(7112..7216) /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 ggatccacta gtccagtgtg gtggaattcg ccaccatgga gaaaaagtgt 721 accctgtatt ttctggttct cttgcctttt tttatgattc ttgttacagc agaattagaa 781 gagagtcctg aggactcaat tcagttggga gttactagaa ataaaatcat gacagctcaa 841 tatgaatgtt accaaaagat tatgcaagac cccattcaac aagcagaagg cgtttactgc 901 aacagaacct gggatggatg gctctgctgg aacgatgttg cagcaggaac tgaatcaatg 961 cagctctgcc ctgattactt tcaggacttt gatccatcag aaaaagttac aaagatctgt 1021 gaccaagatg gaaactggtt tagacatcca gcaagcaaca gaacatggac aaattatacc 1081 cagtgtaatg ttaacaccca cgagaaagtg aagactgcac taaatttgtt ttacctgacc 1141 ataattggac acggattgtc tattgcatca ctgcttatct cgcttggcat attcttttat 1201 ttcaagagcc taagttgcca aaggattacc ttacacaaaa atctgttctt ctcatttgtt 1261 tgtaactctg ttgtaacaat cattcacctc actgcagtgg ccaacaacca ggccttagta 1321 gccacaaatc ctgttagttg caaagtgtcc cagttcattc atctttacct gatgggctgt 1381 aattactttt ggatgctctg tgaaggcatt tacctacaca cactcattgt ggtggccgtg 1441 tttgcagaga agcaacattt aatgtggtat tattttcttg gctggggatt tccactgatt 1501 cctgcttgta tacatgccat tgctagaagc ttatattaca atgacaattg ctggatcagt 1561 tctgataccc atctcctcta cattatccat ggcccaattt gtgctgcttt actggtgaat 1621 ctttttttct tgttaaatat tgtacgcgtt ctcatcacca agttaaaagt tacacaccaa 1681 gcggaatcca atctgtacat gaaagctgtg agagctactc ttatcttggt gccattgctt 1741 ggcattgaat ttgtgctgat tccatggcga cctgaaggaa agattgcaga ggaggtatat 1801 gactacatca tgcacatcct tatgcacttc cagggtcttt tggtctctac cattttctgc 1861 ttctttaatg gagaggttca agcaattctg agaagaaact ggaatcaata caaaatccaa 1921 tttggaaaca gcttttccaa ctcagaagct cttcgtagtg cgtcttacac agtgtcaaca 1981 atcagtgatg gtccaggtta tagtcatgac tgtcctagtg aacacttaaa tggaaaaagc 2041 atccatgata ttgaaaatgt tctcttaaaa ccagaaaatt tatataatct cgaggaacaa 2101 aaactaatat cagaggaaga tttggaacag aaactaatca gtgaagaaga tttagagcaa 2161 aagttaattt ccgaggagga cttatagtga gggcccgata tctcgacaat caacctctgg 2221 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat 2281 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt 2341 tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca 2401 ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg 2461 ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg 2521 aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca 2581 attccgtggt gttgtcgggg aagctgacgt cctttccatg gctgctcgcc tgtgttgcca 2641 cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc 2701 ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc 2761 agacgagtcg gatctccctt tgggccgcct ccccgcctgg aaacggggga ggctaactga 2821 aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa 2881 taaaacgcac gggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc 2941 actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt 3001 ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg 3061 caggccctgc catagcagat ctgcgcagct ggggctctag ggggtatccc cacgcgccct 3121 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg 3181 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg 3241 gctttccccg tcaagctcta aatcggggca tccctttagg gttccgattt agtgctttac 3301 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct 3361 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt 3421 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt 3481 tggggatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt 3541 aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag 3601 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc 3661 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc 3721 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg 3781 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca 3841 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg 3901 tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca 3961 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg 4021 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg 4081 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc 4141 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt 4201 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc 4261 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca 4321 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc 4381 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg 4441 gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct 4501 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc 4561 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc 4621 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta 4681 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt 4741 ctgagcggga ctctggggtt cgcgaaatga ccgaccaagc gacgcccaac ctgccatcac 4801 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg 4861 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca 4921 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa 4981 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt 5041 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt 5101 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa 5161 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac 5221 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg 5281 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc 5341 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat 5401 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca 5461 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc 5521 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc 5581 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg 5641 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta 5701 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg 5761 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac 5821 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag 5881 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat 5941 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat 6001 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc 6061 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt 6121 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct 6181 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt 6241 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc 6301 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac 6361 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat 6421 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg 6481 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata 6541 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta 6601 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt 6661 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag 6721 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa 6781 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc 6841 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt 6901 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc 6961 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta 7021 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa 7081 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca 7141 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac 7201 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag 7261 atctcccgat cccctatggt cgactctcag tacaatctgc tctgatgccg catagttaag 7321 ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta 7381 agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gg //