LOCUS V021053 9866 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V021053 VERSION V021053 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..4513 /label="Npc1(NM_008720)" /note="Npc1(NM_008720)" /gene="Npc1" CDS 4553..4618 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 4642..5230 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 5296..5344 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 5546..5974 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 5988..6317 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 6384..7178 /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 7354..7475 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(7524..7540) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7548..7564 /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(7572..7602) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7617..7638 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7926..8514) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8685..9545) /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(9546..9650) /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 ccatgggtgc gcaccacccg gccctcggcc tgctgctgct 721 gctgctgtgc cctgcgcagg tgttttcgca atcctgtgtt tggtatggag agtgtggaat 781 tgcgactgga gataagaggt acaactgtaa atattctggc ccaccaaaac ccctcccaaa 841 ggacggctat gacttagtgc aggaactctg tccaggactc ttctttgaca atgtcagtct 901 ctgctgtgac attcaacagc ttcagacgct gaagagtaac ctgcagctgc ccctgcagtt 961 cctgtccagg tgtccgtcat gtttttataa cctaatgacc ctgttttgtg agctaacatg 1021 tagcccacac cagagtcagt ttctgaatgt gacagcaact gaggattatt ttgatcctaa 1081 gacacaggag aataaaacaa atgtaaagga attagagtac tttgtcggac agagcttcgc 1141 gaatgcgatg tacaatgcct gccgtgatgt ggaggcgcct tccagtaacg agaaggcctt 1201 aggactcttg tgtgggaggg atgcccgtgc ctgcaatgcc accaactgga ttgagtacat 1261 gttcaataaa gacaacggac aagcgccatt taccatcatt cctgtgtttt cagatctttc 1321 aatccttggg atggagccca tgagaaatgc caccaaaggc tgcaatgagt ctgtagatga 1381 ggtcacgggg ccatgtagct gccaggactg ctccatcgtc tgcggcccca agccccagcc 1441 cccaccccct cctatgccct ggaggatctg gggcttggat gccatgtatg tcatcatgtg 1501 ggtcacctac gtggcatttc tgtttgtgtt ttttggagca ctgttggcag tgtggtgcca 1561 cagaaggcgg tactttgtgt ctgagtacac tcccattgac agtaacatcg ccttttctgt 1621 gaatagcagt gacaaagggg aagcctcatg ctgtgaccca cttggtgcag catttgatga 1681 ctgtctgagg cgcatgttca caaagtgggg ggctttctgt gtccgaaatc ccacctgcat 1741 cattttcttc tcattggcct tcatcactgt gtgctcttct ggcctggtat ttgtccaggt 1801 caccaccaat cctgtagagc tctggtcagc ccctcacagt caggcccgct tggaaaagga 1861 gtactttgac aagcactttg ggcctttctt tcgcacggag cagcttatca tccaggcccc 1921 caacaccagt gttcatatct acgaaccgta ccccgcagga gccgatgtgc ccttcgggcc 1981 tccattgaac aaagagattc tgcaccaggt tctggactta cagatcgcca ttgaaagcat 2041 caccgcatct tacaacaatg aaaccgtgac actgcaggac atctgtgtgg cccccctctc 2101 tccatacaac aagaactgca ccattatgag tgtgttaaat tacttccaga acagccatgc 2161 ggtgctggac agccaagtag gcgacgactt ctatatctac gctgattacc acacacactt 2221 tctgtactgt gtacgggctc ccgcctcctt gaatgatacg agtttgctcc acggtccttg 2281 cctgggtaca tttggaggac cggtgttccc gtggcttgtg ttgggtggct atgatgatca 2341 gaactacaac aatgccaccg cgcttgtgat caccttcccc gtgaataatt actacaatga 2401 cacagagagg ctccagaggg cctgggcctg ggagaaagag tttattagtt ttgtgaaaaa 2461 ctacaagaat ccaaatctga ccatttcttt cactgctgag cgaagcatcg aagatgagct 2521 caatcgggaa agtaacagtg acgtgttcac cgtcatcatc agctacgtcg tgatgtttct 2581 gtacatttcc ctcgccctgg gtcacatcca gagctgcagc aggctcctgg tggattctaa 2641 gatctcgctg ggcattgcgg ggatcctgat cgtgctaagc tcggtggcct gctctctggg 2701 catcttcagc tacatgggga tgccgctgac cctcatcgtc attgaggtca tcccattcct 2761 ggtgctggct gtcggggtgg acaacatctt cattctagtg cagacctacc agagagatga 2821 gcgtcttcag gaggaaacgc tggatcagca gctgggcagg atccttggag aagtggcccc 2881 gaccatgttc ctttcatcct tttctgagac ctcagcattt ttctttgggg cgctgtcctc 2941 gatgccagct gtgcacacct tctctctgtt tgcgggaatg gccgtcctca ttgacttcct 3001 ccttcagatt acctgctttg tgagcctgtt ggggttagat attaagaggc aagagaaaaa 3061 ccatctggac atcctgtgct gtgtcagagg cgctgacgac ggacaaggta gccacgcctc 3121 cgaaagctac ctgtttcgct tcttcaaaaa ctactttgcc ccccttctgc tgaaggactg 3181 gctgaggcca attgtggtag cggtgtttgt gggcgttctg tcattcagtg ttgcggtggt 3241 gaacaaagta gacatcgggt tggatcagtc tctctcaatg ccaaacgatt cgtatgtgat 3301 tgactatttc aaatcactcg ctcagtacct gcactcgggc ccacccgtgt actttgtcct 3361 ggaggaaggc tataactaca gttcacgcaa agggcagaac atggtgtgcg gcggcatggg 3421 ctgtgacaat gactccctgg tgcagcagat atttaacgca gctgagctgg acacctacac 3481 ccgagtaggc ttcgccccct cgtcctggat cgatgactac tttgactggg tctcgccaca 3541 gtcgtcctgc tgcagactct acaacgtcac tcaccagttc tgcaatgctt ctgtgatgga 3601 cccaacctgt gtccgctgca gacctctgac tccagagggt aaacagaggc ctcaggggaa 3661 agaattcatg aaattcctgc ccatgttcct ttctgataac cccaacccca agtgcggcaa 3721 agggggacat gctgcttacg gttcagctgt taacattgtg ggagatgaca cttacattgg 3781 ggccacttac ttcatgacct accacaccat acttaagacc tccgctgact atactgatgc 3841 catgaagaaa gctcggctaa tagccagtaa catcacggaa accatgcgtt ctaaggggag 3901 tgactaccgc gtattccctt acagtgtgtt ctacgtcttc tatgaacagt acctgaccat 3961 tattgatgac accatcttta acctcagtgt gtctctgggc tccatatttc tggtgacctt 4021 ggtggttctg ggctgtgagc tgtggtctgc ggtcatcatg tgtatcacca tagccatgat 4081 cctggtcaac atgttcggtg tcatgtggct gtggggcatc agtctgaatg cggtctcctt 4141 ggtcaacttg gtgatgagct gcggcatttc tgtggagttc tgcagccata taacgagagc 4201 attcaccatg agtaccaaag gaagccgagt gagccgggcg gaagaggcac tggcccacat 4261 gggtagttct gtattcagtg gaatcacact tacgaaattt ggagggatcg tggtgttagc 4321 ctttgccaaa tctcaaattt ttgagatatt ttacttcagg atgtacttag ccatggtctt 4381 actcggagcc actcatggac taatatttct tcccgtctta ctcagttaca tagggccgtc 4441 ggtgaataaa gctaaaagac acaccactta cgagcgctac agagggacag agagagaacg 4501 gctcctcaat tttgaattct gcagatatcc agcacagtgg cggccgctcg aggactacaa 4561 agaccatgac ggtgattata aagatcatga catcgactac aaggatgacg atgacaagta 4621 gtgagggccc gatatctcga caatcaacct ctggattaca aaatttgtga aagattgact 4681 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg 4741 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg 4801 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg 4861 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg 4921 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc 4981 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaagctg 5041 acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc 5101 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct 5161 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc 5221 gcctccccgc ctggaaacgg gggaggctaa ctgaaacacg gaaggagaca ataccggaag 5281 gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacgggtgt tgggtcgttt 5341 gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc 5401 cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt 5461 gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc agatctgcgc 5521 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt 5581 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc 5641 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg 5701 ggcatccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat 5761 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg 5821 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct 5881 atctcggtct attcttttga tttataaggg attttgggga tttcggccta ttggttaaaa 5941 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag 6001 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt 6061 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca 6121 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa 6181 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag 6241 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag 6301 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag 6361 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg 6421 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg 6481 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc 6541 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga 6601 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc 6661 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag 6721 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat 6781 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg 6841 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca 6901 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct 6961 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg 7021 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg 7081 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc 7141 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgcgaa 7201 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc 7261 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc 7321 ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt 7381 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct 7441 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct 7501 agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc 7561 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga 7621 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg 7681 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg 7741 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg 7801 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga 7861 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg 7921 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag 7981 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc 8041 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg 8101 ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt 8161 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc 8221 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc 8281 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg 8341 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca 8401 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc 8461 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat 8521 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt 8581 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt 8641 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc 8701 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc 8761 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata 8821 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg 8881 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc 8941 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct 9001 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa 9061 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt 9121 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca 9181 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac 9241 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca 9301 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt 9361 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc 9421 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca 9481 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata 9541 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc 9601 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc 9661 cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtcgactc 9721 tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt 9781 tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg 9841 acaattgcat gaagaatctg cttagg //