LOCUS V025881 8022 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V025881 VERSION V025881 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..2702 /label="RILP(NM_031430)" /note="RILP(NM_031430)" /gene="RILP" CDS 2709..2774 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 2798..3386 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 3452..3500 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 3702..4130 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 4144..4473 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4540..5334 /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 5510..5631 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5680..5696) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5704..5720 /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(5728..5758) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5773..5794 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6082..6670) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6841..7701) /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(7702..7806) /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 tggagcccag gagggcggcg cccggggtgc ctggctgggg gtctcgggag gccgcggggt 1561 cggcatcggc cgcggagctt gtgtaccatc tagccggggc cctgggcact gagctgcagg 1621 atctggcgcg ccgtttcggg ccggaggcgg cggccgggct ggtgccgcta gtggtgcggg 1681 cgctggagct cttggaacag gctgccgtgg ggcccgcccc ggactcgctg caggtgtcgg 1741 cgcagccggc ggagcaggag ctgcggcggc tgcgggagga gaacgagcgc ctccgcaggg 1801 agctgcgcgc ggggccacag gaggagcgcg cgctgctgcg gcagctcaag gaggtcacgg 1861 accgacagcg ggacgaactc cgggcgcaca accgcgacct gcggcagcgc ggccaggaga 1921 ccgaggcgtt gcaggagcag ctgcagcgcc tcctgctggt gaacgctgag ctgcggcaca 1981 agctggcggc catgcagacc cagctgcgcg ccgcgcagga ccgcgagagg gagcgccagc 2041 agcctggcga agccgcgact ccgcaggcta aagagcgagc gcgggggcag gccgggcggc 2101 ccgggcacca gcacggacag gagcccgaat gggcgaccgc cggcgcaggc gccccaggga 2161 accctgagga cccggcggag gccgcgcagc agctcgggcg cccctcggag gcagggcagt 2221 gccgcttcag tcgggaggag tttgagcaga tccttcagga gcggaatgaa ctcaaagcca 2281 aagtgttcct gctcaaggag gaactggcct acttccagcg ggagctgctc acagaccacc 2341 gggtccccgg ccttctgctc gaggccatga aggtggctgt ccggaagcag cggaagaaga 2401 tcaaggccaa gatgttaggg acaccagagg aagcagagag cagtgaggat gaggctggcc 2461 catggatcct gctctccgat gacaagggag accatccccc acccccggag tccaaaatac 2521 agagtttctt tggcctatgg tatcggggta aagctgaatc ctctgaggat gagaccagca 2581 gccctgcacc cagcaagcta gggggagaag aggaggccca accacagtct ccagctcctg 2641 atccgccctg ttctgccctc cacgaacacc tttgtctggg ggcctcagcc gccccagagg 2701 ccctcgagga ctacaaagac catgacggtg attataaaga tcatgacatc gactacaagg 2761 atgacgatga caagtagtga gggcccgata tctcgacaat caacctctgg attacaaaat 2821 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc 2881 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt 2941 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg 3001 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg 3061 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc 3121 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt 3181 gttgtcgggg aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct 3241 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg 3301 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg 3361 gatctccctt tgggccgcct ccccgcctgg aaacggggga ggctaactga aacacggaag 3421 gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac 3481 gggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga 3541 taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca 3601 ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc 3661 catagcagat ctgcgcagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc 3721 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct 3781 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg 3841 tcaagctcta aatcggggca tccctttagg gttccgattt agtgctttac ggcacctcga 3901 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt 3961 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg 4021 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tggggatttc 4081 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg 4141 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa 4201 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc 4261 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg 4321 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt 4381 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga 4441 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt 4501 ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg 4561 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag 4621 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt 4681 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta 4741 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg 4801 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt 4861 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat 4921 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg 4981 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca 5041 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc 5101 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc 5161 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat 5221 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc 5281 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga 5341 ctctggggtt cgcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga 5401 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg 5461 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat 5521 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt 5581 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg 5641 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg 5701 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc 5761 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt 5821 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg 5881 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt 5941 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc 6001 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa 6061 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa 6121 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc 6181 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc 6241 cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag 6301 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga 6361 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc 6421 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac 6481 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg 6541 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca 6601 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa 6661 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa 6721 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt 6781 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag 6841 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat 6901 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc 6961 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa 7021 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca 7081 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa 7141 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt 7201 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc 7261 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact 7321 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc 7381 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg 7441 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct 7501 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc 7561 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag 7621 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac 7681 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg 7741 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt 7801 tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat 7861 cccctatggt cgactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg 7921 ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca 7981 aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gg //