LOCUS V027495 9485 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V027495 VERSION V027495 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 775..3411 /label="GRM3(NM_000840)" /note="GRM3(NM_000840)" /gene="GRM3" CDS 3427..3483 /codon_start=1 /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /product="2A peptide from porcine teschovirus-1 polyprotein" /transl_table=1 /translation="ATNFSLLKQAGDVEENPGP" /label="P2A" CDS 3484..4194 /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" misc_feature 4261..4849 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 4915..4963 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 5165..5593 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 5607..5936 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 6003..6797 /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 6973..7094 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(7143..7159) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7167..7183 /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(7191..7221) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7236..7257 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7545..8133) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8304..9164) /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(9165..9269) /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 gctcgtttag tgaaccgtca gatcgcctgg 661 agacgccatc cacgctgttt tgacctccat agaagacacc gggaccgatc cagcctccgg 721 actctagagc tagcgtttaa acttaagctt ggtaccgagc tcggatccgc caccatgaag 781 atgttgacaa gactgcaagt tcttacctta gctttgtttt caaagggatt tttactctct 841 ttaggggacc ataactttct aaggagagag attaaaatag aaggtgacct tgttttaggg 901 ggcctgtttc ctattaacga aaaaggcact ggaactgaag aatgtgggcg aatcaatgaa 961 gaccgaggga ttcaacgcct ggaagccatg ttgtttgcta ttgatgaaat caacaaagat 1021 gattacttgc taccaggagt gaagttgggt gttcacattt tggatacatg ttcaagggat 1081 acctatgcat tggagcaatc actggagttt gtcagggcat ctttgacaaa agtggatgaa 1141 gctgagtata tgtgtcctga tggatcctat gccattcaag aaaacatccc acttctcatt 1201 gcaggggtca ttggtggctc ttatagcagt gtttccatac aggtggcaaa cctgctgcgg 1261 ctcttccaga tccctcagat cagctacgca tccaccagcg ccaaactcag tgataagtcg 1321 cgctatgatt actttgccag gaccgtgccc cccgacttct accaggccaa agccatggct 1381 gagatcttgc gcttcttcaa ctggacctac gtgtccacag tagcctccga gggtgattac 1441 ggggagacag ggatcgaggc cttcgagcag gaagcccgcc tgcgcaacat ctgcatcgct 1501 acggcggaga aggtgggccg ctccaacatc cgcaagtcct acgacagcgt gatccgagaa 1561 ctgttgcaga agcccaacgc gcgcgtcgtg gtcctcttca tgcgcagcga cgactcgcgg 1621 gagctcattg cagccgccag ccgcgccaat gcctccttca cctgggtggc cagcgacggc 1681 tggggcgcgc aggagagcat catcaagggc agcgagcatg tggcctacgg cgccatcacc 1741 ctggagctgg cctcccagcc tgtccgccag ttcgaccgct acttccagag cctcaacccc 1801 tacaacaacc accgcaaccc ctggttccgg gacttctggg agcaaaagtt tcagtgcagc 1861 ctccagaaca aacgcaacca caggcgcgtc tgcgacaagc acctggccat cgacagcagc 1921 aactacgagc aagagtccaa gatcatgttt gtggtgaacg cggtgtatgc catggcccac 1981 gctttgcaca aaatgcagcg caccctctgt cccaacacta ccaagctttg tgatgctatg 2041 aagatcctgg atgggaagaa gttgtacaag gattacttgc tgaaaatcaa cttcacggct 2101 ccattcaacc caaataaaga tgcagatagc atagtcaagt ttgacacttt tggagatgga 2161 atggggcgat acaacgtgtt caatttccaa aatgtaggtg gaaagtattc ctacttgaaa 2221 gttggtcact gggcagaaac cttatcgcta gatgtcaact ctatccactg gtcccggaac 2281 tcagtcccca cttcccagtg cagcgacccc tgtgccccca atgaaatgaa gaatatgcaa 2341 ccaggggatg tctgctgctg gatttgcatc ccctgtgaac cctacgaata cctggctgat 2401 gagtttacct gtatggattg tgggtctgga cagtggccca ctgcagacct aactggatgc 2461 tatgaccttc ctgaggacta catcaggtgg gaagacgcct gggccattgg cccagtcacc 2521 attgcctgtc tgggttttat gtgtacatgc atggttgtaa ctgtttttat caagcacaac 2581 aacacaccct tggtcaaagc atcgggccga gaactctgct acatcttatt gtttggggtt 2641 ggcctgtcat actgcatgac attcttcttc attgccaagc catcaccagt catctgtgca 2701 ttgcgccgac tcgggctggg gagttccttc gctatctgtt actcagccct gctgaccaag 2761 acaaactgca ttgcccgcat cttcgatggg gtcaagaatg gcgctcagag gccaaaattc 2821 atcagcccca gttctcaggt tttcatctgc ctgggtctga tcctggtgca aattgtgatg 2881 gtgtctgtgt ggctcatcct ggaggcccca ggcaccagga ggtataccct tgcagagaag 2941 cgggaaacag tcatcctaaa atgcaatgtc aaagattcca gcatgttgat ctctcttacc 3001 tacgatgtga tcctggtgat cttatgcact gtgtacgcct tcaaaacgcg gaagtgccca 3061 gaaaatttca acgaagctaa gttcataggt tttaccatgt acaccacgtg catcatctgg 3121 ttggccttcc tccctatatt ttatgtgaca tcaagtgact acagagtgca gacgacaacc 3181 atgtgcatct ctgtcagcct gagtggcttt gtggtcttgg gctgtttgtt tgcacccaag 3241 gttcacatca tcctgtttca accccagaag aatgttgtca cacacagact gcacctcaac 3301 aggttcagtg tcagtggaac tgggaccaca tactctcagt cctctgcaag cacgtatgtg 3361 ccaacggtgt gcaatgggcg ggaagtcctc gactccacca cctcatctct ggaattcgga 3421 agcggagcta ctaacttcag cctgctgaag caggctggcg acgtggagga gaaccctgga 3481 cctatggtga gcaagggcga ggaggataac atggccatca tcaaggagtt catgcgcttc 3541 aaggtgcaca tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag 3601 ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg tggccccctg 3661 cccttcgcct gggacatcct gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag 3721 caccccgccg acatccccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag 3781 cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag 3841 gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc 3901 gtaatgcaga agaagaccat gggctgggag gcctcctccg agcggatgta ccccgaggac 3961 ggcgccctga agggcgagat caagcagagg ctgaagctga aggacggcgg ccactacgac 4021 gctgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacaac 4081 gtcaacatca agttggacat cacctcccac aacgaggact acaccatcgt ggaacagtac 4141 gaacgcgccg agggccgcca ctccaccggc ggcatggacg agctgtacaa gtaactcgag 4201 tctagagggc cccttaaggg ttcgatccct accggttagt aatgagtttg atatctcgac 4261 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct 4321 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt 4381 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg 4441 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact 4501 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct 4561 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg 4621 ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc atggctgctc 4681 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc 4741 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt 4801 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc tggaaacggg 4861 ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa 4921 taaaaagaca gaataaaacg cacgggtgtt gggtcgtttg ttcataaacg cggggttcgg 4981 tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc 5041 gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc 5101 aacgtcgggg cggcaggccc tgccatagca gatctgcgca gctggggctc tagggggtat 5161 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg 5221 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc 5281 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg gcatcccttt agggttccga 5341 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt 5401 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat 5461 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat 5521 ttataaggga ttttggggat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa 5581 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct 5641 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa 5701 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa 5761 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt 5821 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct 5881 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc 5941 tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc 6001 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat 6061 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt 6121 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac 6181 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg 6241 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc 6301 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa 6361 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc 6421 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg 6481 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg 6541 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa 6601 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg 6661 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct 6721 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc 6781 ttgacgagtt cttctgagcg ggactctggg gttcgcgaaa tgaccgacca agcgacgccc 6841 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga 6901 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc 6961 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc 7021 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc 7081 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca 7141 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga 7201 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt 7261 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga 7321 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc 7381 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg 7441 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc 7501 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc 7561 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga 7621 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc 7681 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa 7741 tgctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg 7801 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc 7861 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga 7921 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact 7981 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt 8041 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag 8101 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg 8161 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa 8221 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata 8281 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg 8341 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata 8401 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg 8461 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct 8521 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt 8581 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc 8641 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga 8701 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt 8761 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc 8821 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa 8881 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca 8941 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca 9001 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct 9061 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc 9121 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa 9181 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt 9241 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc 9301 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg 9361 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 9421 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 9481 ttagg //