LOCUS V041381 8019 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V041381 VERSION V041381 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 744..1451 /codon_start=1 /note="mammalian codon-optimized" /product="monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)" /transl_table=1 /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA EGRHSTGGMDELYK" /label="mCherry" CDS 1500..2699 /label="KRT19(NM_002276)" /note="KRT19(NM_002276)" /gene="KRT19" CDS 2706..2771 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 2795..3383 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 3449..3497 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 3699..4127 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 4141..4470 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4537..5331 /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 5507..5628 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5677..5693) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5701..5717 /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(5725..5755) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5770..5791 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6079..6667) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6838..7698) /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(7699..7803) /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 tggtaccgcc accatggtga gcaagggcga ggaggataac atggccatca 781 tcaaggagtt catgcgcttc aaggtgcaca tggagggctc cgtgaacggc cacgagttcg 841 agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg 901 tgaccaaggg tggccccctg cccttcgcct gggacatcct gtcccctcag ttcatgtacg 961 gctccaaggc ctacgtgaag caccccgccg acatccccga ctacttgaag ctgtccttcc 1021 ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga 1081 cccaggactc ctccctgcag gacggcgagt tcatctacaa ggtgaagctg cgcggcacca 1141 acttcccctc cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctcctccg 1201 agcggatgta ccccgaggac ggcgccctga agggcgagat caagcagagg ctgaagctga 1261 aggacggcgg ccactacgac gctgaggtca agaccaccta caaggccaag aagcccgtgc 1321 agctgcccgg cgcctacaac gtcaacatca agttggacat cacctcccac aacgaggact 1381 acaccatcgt ggaacagtac gaacgcgccg agggccgcca ctccaccggc ggcatggacg 1441 agctgtacaa gtggatcgga ggagggggaa gcggtggagg cggtagtgag ctcggatcca 1501 tgacttccta cagctatcgc cagtcgtcgg ccacgtcgtc cttcggaggc ctgggcggcg 1561 gctccgtgcg ttttgggccg ggggtcgcct ttcgcgcgcc cagcattcac gggggctccg 1621 gcggccgcgg cgtatccgtg tcctccgccc gctttgtgtc ctcgtcctcc tcgggggcct 1681 acggcggcgg ctacggcggc gtcctgaccg cgtccgacgg gctgctggcg ggcaacgaga 1741 agctaaccat gcagaacctc aacgaccgcc tggcctccta cctggacaag gtgcgcgccc 1801 tggaggcggc caacggcgag ctagaggtga agatccgcga ctggtaccag aagcaggggc 1861 ctgggccctc ccgcgactac agccactact acacgaccat ccaggacctg cgggacaaga 1921 ttcttggtgc caccattgag aactccagga ttgtcctgca gatcgacaat gcccgtctgg 1981 ctgcagatga cttccgaacc aagtttgaga cggaacaggc tctgcgcatg agcgtggagg 2041 ccgacatcaa cggcctgcgc agggtgctgg atgagctgac cctggccagg accgacctgg 2101 agatgcagat cgaaggcctg aaggaagagc tggcctacct gaagaagaac catgaggagg 2161 aaatcagtac gctgaggggc caagtgggag gccaggtcag tgtggaggtg gattccgctc 2221 cgggcaccga tctcgccaag atcctgagtg acatgcgaag ccaatatgag gtcatggccg 2281 agcagaaccg gaaggatgct gaagcctggt tcaccagccg gactgaagaa ttgaaccggg 2341 aggtcgctgg ccacacggag cagctccaga tgagcaggtc cgaggttact gacctgcggc 2401 gcacccttca gggtcttgag attgagctgc agtcacagct gagcatgaaa gctgccttgg 2461 aagacacact ggcagaaacg gaggcgcgct ttggagccca gctggcgcat atccaggcgc 2521 tgatcagcgg tattgaagcc cagctgggcg atgtgcgagc tgatagtgag cggcagaatc 2581 aggagtacca gcggctcatg gacatcaagt cgcggctgga gcaggagatt gccacctacc 2641 gcagcctgct cgagggacag gaagatcact acaacaattt gtctgcctcc aaggtcctcc 2701 tcgaggacta caaagaccat gacggtgatt ataaagatca tgacatcgac tacaaggatg 2761 acgatgacaa gtagtgaggg cccgatatct cgacaatcaa cctctggatt acaaaatttg 2821 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc 2881 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta 2941 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt 3001 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca 3061 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc 3121 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt 3181 gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg 3241 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg 3301 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat 3361 ctccctttgg gccgcctccc cgcctggaaa cgggggaggc taactgaaac acggaaggag 3421 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggg 3481 tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac 3541 cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc 3601 cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat 3661 agcagatctg cgcagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt 3721 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc 3781 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca 3841 agctctaaat cggggcatcc ctttagggtt ccgatttagt gctttacggc acctcgaccc 3901 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt 3961 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac 4021 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgg ggatttcggc 4081 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat 4141 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc 4201 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga 4261 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc 4321 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt 4381 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga 4441 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc 4501 ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac 4561 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca 4621 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt 4681 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg 4741 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga 4801 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct 4861 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg 4921 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg 4981 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc 5041 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat 5101 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac 5161 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt 5221 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct 5281 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc 5341 tggggttcgc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc 5401 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat 5461 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc 5521 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt 5581 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat 5641 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa 5701 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg 5761 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca 5821 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg 5881 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg 5941 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg 6001 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa 6061 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg 6121 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc 6181 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc 6241 ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc 6301 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg 6361 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc 6421 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga 6481 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc 6541 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac 6601 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg 6661 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc 6721 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa 6781 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta 6841 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt 6901 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag 6961 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca 7021 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc 7081 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt 7141 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag 7201 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt 7261 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat 7321 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt 7381 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc 7441 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat 7501 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag 7561 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt 7621 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg 7681 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta 7741 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc 7801 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc 7861 ctatggtcga ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc 7921 cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg 7981 caaggcttga ccgacaattg catgaagaat ctgcttagg //