LOCUS V036171 7442 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V036171 VERSION V036171 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..1368 /label="RGS8(NM_033345)" /note="RGS8(NM_033345)" /gene="RGS8" CDS 1384..1440 /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 1441..2151 /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 2218..2806 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 2872..2920 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 3122..3550 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3564..3893 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3960..4754 /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 4930..5051 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5100..5116) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5124..5140 /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(5148..5178) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5193..5214 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5502..6090) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6261..7121) /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(7122..7226) /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 caccatgtgg 781 aacaccttaa cccgaagcct ctctgaccat ccagttggca aagaccctca ggccatgagg 841 actggccaaa gacagaacaa agggatgagg actcgactgg gatgcctgtc tcacaagtca 901 gactcgtgta gtgatttcac agctattctt ccagacaaac ccaaccgcgc tctcaagaga 961 ttatcgacag aagaagctac gaggtgggca gattcctttg atgtgcttct ctctcataag 1021 tatggggtgg ctgcattccg tgccttcttg aagacggagt tcagtgagga gaacctggaa 1081 ttctggttgg cctgtgagga gttcaagaag accaggtcaa ctgcaaaact ggtctctaag 1141 gcccatagga tctttgagga gtttgtggat gtgcaggctc cacgggaggt aaacattgac 1201 ttccagaccc gagaagccac gaggaagaac ctgcaggagc catccctgac ttgctttgac 1261 caagcccaag gaaaagtaca cagcctcatg gagaaagact cttaccccag gttcctgagg 1321 tccaaaatgt acttagatct gctgtcccaa agccagagga ggctcagtga attcggaagc 1381 ggagctacta acttcagcct gctgaagcag gctggcgacg tggaggagaa ccctggacct 1441 atggtgagca agggcgagga ggataacatg gccatcatca aggagttcat gcgcttcaag 1501 gtgcacatgg agggctccgt gaacggccac gagttcgaga tcgagggcga gggcgagggc 1561 cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc 1621 ttcgcctggg acatcctgtc ccctcagttc atgtacggct ccaaggccta cgtgaagcac 1681 cccgccgaca tccccgacta cttgaagctg tccttccccg agggcttcaa gtgggagcgc 1741 gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc aggactcctc cctgcaggac 1801 ggcgagttca tctacaaggt gaagctgcgc ggcaccaact tcccctccga cggccccgta 1861 atgcagaaga agaccatggg ctgggaggcc tcctccgagc ggatgtaccc cgaggacggc 1921 gccctgaagg gcgagatcaa gcagaggctg aagctgaagg acggcggcca ctacgacgct 1981 gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc ctacaacgtc 2041 aacatcaagt tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgaa 2101 cgcgccgagg gccgccactc caccggcggc atggacgagc tgtacaagta actcgagtct 2161 agagggcccc ttaagggttc gatccctacc ggttagtaat gagtttgata tctcgacaat 2221 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct 2281 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg 2341 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg 2401 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt 2461 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt 2521 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg 2581 ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg gctgctcgcc 2641 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat 2701 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc 2761 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg aaacggggga 2821 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa 2881 aaagacagaa taaaacgcac gggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc 2941 cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt 3001 tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac 3061 gtcggggcgg caggccctgc catagcagat ctgcgcagct ggggctctag ggggtatccc 3121 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc 3181 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc 3241 acgttcgccg gctttccccg tcaagctcta aatcggggca tccctttagg gttccgattt 3301 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg 3361 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt 3421 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta 3481 taagggattt tggggatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt 3541 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc 3601 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt 3661 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca 3721 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc 3781 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg 3841 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc 3901 cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca 3961 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg 4021 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag 4081 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc 4141 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc 4201 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg 4261 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc 4321 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca 4381 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag 4441 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg 4501 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg 4561 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca 4621 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc 4681 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg 4741 acgagttctt ctgagcggga ctctggggtt cgcgaaatga ccgaccaagc gacgcccaac 4801 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc 4861 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc 4921 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca 4981 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc 5041 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg 5101 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc 5161 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg 5221 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc 5281 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact 5341 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta 5401 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag 5461 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc 5521 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta 5581 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg 5641 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc 5701 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac 5761 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac 5821 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg 5881 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga 5941 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt 6001 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag 6061 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct 6121 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg 6181 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat 6241 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc 6301 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg 6361 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct 6421 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca 6481 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg 6541 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg 6601 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc 6661 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag 6721 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg 6781 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag 6841 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat 6901 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg 6961 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca 7021 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca 7081 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat 7141 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag 7201 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac 7261 ggatcgggag atctcccgat cccctatggt cgactctcag tacaatctgc tctgatgccg 7321 catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga 7381 gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta 7441 gg //