LOCUS V044387 9246 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V044387 VERSION V044387 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" promoter 678..696 /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" CDS 756..3926 /label="Herc4(NM_030114)" /note="Herc4(NM_030114)" /gene="Herc4" CDS 3933..3998 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 4022..4610 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 4676..4724 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 4926..5354 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 5368..5697 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 5764..6558 /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 6734..6855 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(6904..6920) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6928..6944 /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(6952..6982) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6997..7018 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7306..7894) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8065..8925) /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(8926..9030) /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 gctctctggc taactagaga acccactgct 661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta 721 aacttaagct tggtaccgag ctcgaattcg ccaccatgtt gtgctggggc aatgcatcct 781 atggacaact aggtttggga ggaattgatg aagaaattgt actagagccc aggagaagtg 841 actttttcgt gaacaaaaag gtccgagatg taggctgtgg actcaggcac actgtgtttg 901 tcctggatga tgggactgtg tacacatgtg gatgtaatga tctaggacag ctaggtcatg 961 aaaagtccag aaagaaacca gagcaggttg ttgccctgga tgcccagaat atcgtagctg 1021 ttgcgtgtgg agaagctcac acgttagcgc tgaatgacaa gggccaggtg tatgcttggg 1081 gtctcgactc tgacggacag cttggcctac agggatcaga ggaatgtatc agagtaccca 1141 gaaatattaa aagtctttcg gatatccaga tagtacaggt tgcgtgtggt tactatcatt 1201 cgcttgcact ttctaaagca agtgaagttt tctgttgggg acagaataaa tatggccagc 1261 tgggtctagg cattgattgt caaaagcaaa cttcaccaca gctgattaag tctttgcttg 1321 gaataccatt catgcaagtc gcagcaggag gagcccatag ttttgtactc accctttccg 1381 gagctatctt tggatgggga cgtaacaaat ttggtcagct aggtcttaat gatgaaaatg 1441 ataggtatgt tcctaattta ctaaagtcac taagatctca gaaaatagtt tatatttgtt 1501 gcggagaaga tcatactgct gcattaacca aggaaggtgg agtgtttacc tttggagctg 1561 ggggctatgg ccagctgggt cataattcta ccagtcatga gataaatccc aggaaagttt 1621 ttgaactcat gggaagcatt gttactcagg tcgcttgtgg aaggcagcat acctctgcat 1681 ttgttccttc ttcaggacga atttactcct ttggtcttgg tggtaatggg cagttgggaa 1741 ctggttcaac cagcaacaga aaaagccctt tcactgtaaa aggaaactgg ttttcttata 1801 atggacagtg tccacaagat attggttctg aagactattt ctgtgtcaaa agaattttct 1861 caggtggaga tcaaagcttt tcacattact ctagtcctca gaactgtgga cctccagatg 1921 actttagatg ctccgaccct tccaagcaga tctggacagt gaacgaagct ctgattcaga 1981 aatggctcag ctacccctct ggaaggttcc ctgtggagat agccaatgag atagatggaa 2041 cattttcttc atctggctgc ctaaatggaa gttttctggc tattagcaat gacgatcact 2101 atagaacagg taccagattt tcaggggttg atatgaacgc tgctagactc ttattccaca 2161 aacttataca acctgatcat cctcagatat ctcagcaggt ggcagctagt ttggaaaaga 2221 atctaatccc taaactgact agttccctac ctgatgttga agctctgagg ttttatctta 2281 ccctgccaga gtgtcccctg atgagtgact gcaacaattt cacaacaata gcaatcccct 2341 ttggtacagc tcttgtgaac ctagaaaagg caccactgaa agtacttgaa aactggtggt 2401 cagtacttga acctccacta ttcctcaaga tagtagaact ttttaaggaa gttgtggtac 2461 atcttttgaa actctacaag atcggcattc ccccttctga aagaagaatt ttcaacagtt 2521 ttcttcatac cgcattaaag gttttagaaa tattgcatag ggtaaatgag aaaacaggac 2581 aacttattca atatgacaag ttttacatcc atgaagtcca agagctgata gacataagga 2641 atgactatat caactgggtc cagcagcaag cctatggagt ggatgtcagc catggagtaa 2701 ctgagttggc agatatcccg gtgacaattt gtacatatcc atttgtattt gatgcccagg 2761 caaaaactac tctgctccag acagatgcag tcttgcagat gcagatggcc attgaccagg 2821 cccacaggca gaatgtctcc tctctttttc tcccagtgat cgagtctgtg aatccttgct 2881 taattctagt tgttcgcaga gagaatattg taggagatgc gatggaagtc ctcaggaaaa 2941 ccaagaatat agattataaa aagccactca aggttatatt tgttggagaa gatgctgttg 3001 atgcaggagg tgttcgcaaa gaatttttct tgcttatcat gagggaattg ttggatccta 3061 aatacggcat gtttcgatat tatgaagatt ccaggctcat ttggttttca gataagacat 3121 ttgaagacag tgatttgttc cacttgatcg gtgttatctg tggattagca atttataatt 3181 ttaccattgt ggacctccat ttccctttgg ctttatataa gaaactgctg aaaaggaagc 3241 cgtccctgga tgatctgaaa gagctgatgc cagccgtcgg gagaagcatg cagcaattgc 3301 tggactatcc agaagatgac atagaggaaa cattttgtct aaactttacg atcacagttg 3361 agaacttcgg tgcaacagaa gtaaaagagc tggttctgaa cggggcagac actgctgtta 3421 atagacagaa tcggcaggag tttgttgatg cctatgtgga ttacatattc aataaatcag 3481 tggcatcttt atttgatgct ttccatgcgg gctttcataa ggtctgtgga ggaaaagttc 3541 ttctgctctt ccagcctaat gaattacaag ctatggttat tgggaataca aattatgatt 3601 ggaaagaact ggagaagaat acagaataca aaggagagta ttgggcagac catcccacga 3661 taaaaatttt ttgggaagtg tttcatgaat taccactaga gaagaagaaa caatttctat 3721 tatttttgac aggtagtgat cggattccca ttcttggtat gaagagtctg aaactagtca 3781 tccagtcaac aggaggtggt gagagctatc tcccggtttc ccatacctgt tttaatcttc 3841 tagatctccc aaaatataca gagaaagaaa ccctccgatg taaactgatc caagctattg 3901 atcacaatga aggcttcagt ttaataggat ccgactacaa agaccatgac ggtgattata 3961 aagatcatga catcgactac aaggatgacg atgacaagta gtgagggccc gatatctcga 4021 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc 4081 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg 4141 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt 4201 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac 4261 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc 4321 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct 4381 gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct 4441 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct 4501 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct 4561 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggaaacgg 4621 gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca 4681 ataaaaagac agaataaaac gcacgggtgt tgggtcgttt gttcataaac gcggggttcg 4741 gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg 4801 cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc 4861 caacgtcggg gcggcaggcc ctgccatagc agatctgcgc agctggggct ctagggggta 4921 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt 4981 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct 5041 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg ggcatccctt tagggttccg 5101 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag 5161 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa 5221 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga 5281 tttataaggg attttgggga tttcggccta ttggttaaaa aatgagctga tttaacaaaa 5341 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc 5401 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga 5461 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca 5521 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat 5581 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc 5641 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag 5701 ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt 5761 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta 5821 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg 5881 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa 5941 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct 6001 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg 6061 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca 6121 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat 6181 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac 6241 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc 6301 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa 6361 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag 6421 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc 6481 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt 6541 cttgacgagt tcttctgagc gggactctgg ggttcgcgaa atgaccgacc aagcgacgcc 6601 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg 6661 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt 6721 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat 6781 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact 6841 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc 6901 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg 6961 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat 7021 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg 7081 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct 7141 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc 7201 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg 7261 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg 7321 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg 7381 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac 7441 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca 7501 atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt 7561 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc 7621 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag 7681 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac 7741 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt 7801 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa 7861 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg 7921 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa 7981 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat 8041 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc 8101 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat 8161 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc 8221 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc 8281 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag 8341 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg 8401 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg 8461 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag 8521 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt 8581 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga 8641 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc 8701 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc 8761 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc 8821 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc 8881 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca 8941 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat 9001 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt 9061 cgacggatcg ggagatctcc cgatccccta tggtcgactc tcagtacaat ctgctctgat 9121 gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc 9181 gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg 9241 cttagg //