LOCUS Exported 8663 bp ds-DNA circular SYN 10-JUN-2019 DEFINITION . ACCESSION . VERSION . KEYWORDS pMRX-INU-3xFLAG-CCPG1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8663) AUTHORS Noboru Mizushima TITLE Direct Submission JOURNAL Exported Monday, June 10, 2019 from SnapGene 3.3.4 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8663 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 1..380 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" /note="color: #ffffff; direction: RIGHT" misc_feature 381..676 /label=LTR /note="color: #ffe4c4" misc_feature 739..1096 /label=MMLV Psi /note="packaging signal of Moloney murine leukemia virus (MMLV)" /note="color: #ffe4c4" CDS 1161..1577 /label=gag (truncated) /note="truncated Moloney murine leukemia virus (MMLV) gag gene lacking the start codon" /note="color: #ffff99; direction: RIGHT" misc_feature 1587..1961 /label=pol region /note="Moloney murine leukemia virus (MMLV) pol region containing the splice acceptor site" /note="color: #ffe4c4 Cleavage site after base 1941" CDS 1994..2062 /codon_start=1 /label=3xFLAG /note="color: #003300" /translation="MDYKDHDGDYKDHDIDYKDDDDK" CDS 2063..2074 /codon_start=1 /label=MCS /note="color: #000080" /translation="GSGS" misc_feature 2081..4354 /label=CCPG1 /note="color: #ff00ff" CDS 2120..2131 /codon_start=1 /label=LIR /note="color: #993366" /translation="WTVI" CDS 2144..2161 /codon_start=1 /label=FIR /note="color: #993366" /translation="SDIEML" CDS 2390..2407 /codon_start=1 /label=FIR /note="color: #993366" /translation="SDIVTL" misc_feature 2711..2779 /label=TM /note="color: #a6acb3" misc_feature 4395..4947 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" /note="color: #ffffff; direction: RIGHT" CDS 4965..5759 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /note="color: #ccffcc" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" LTR 5989..6458 /note="long terminal repeat from Moloney murine leukemia virus" /note="color: #ffe4c4" rep_origin complement(6759..7347) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /note="color: #ffff99" CDS complement(7511..8371) /codon_start=1 /label=AmpR /note="color: #ccffcc" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" ORIGIN 1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc 61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca 121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga 181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc 241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct 301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat 361 tagtcatcgc tattaccatg ctagagaacc atcagatgtt tccagggtgc cccaaggacc 421 tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc 481 gcgcttctgc tccccgagct caataaaaga gcccacaacc cctcactcgg cgcgccagtc 541 ctccgattga ctgagtcgcc cgggtacccg tgtatccaat aaaccctctt gcagttgcat 601 ccgacttgtg gtctcgctgt tccttgggag ggtctcctct gagtgattga ctacccgtca 661 gcgggggtct ttcatttggg ggctcgtccg ggatcgggag acccctgccc agggaccacc 721 gacccaccac cgggaggtaa gctggccagc aacttatctg tgtctgtccg attgtctagt 781 gtctatgact gattttatgc gcctgcgtcg gtactagtta gctaactagc tctgtatctg 841 gcggacccgt ggtggaactg acgagttcgg aacacccggc cgcaaccctg ggagacgtcc 901 cagggacttc gggggccgtt tttgtggccc gacctgagtc caaaaatccc gatcgttttg 961 gactctttgg tgcacccccc ttagaggagg gatatgtggt tctggtagga gacgagaacc 1021 taaaacagtt cccgcctccg tctgaatttt tgctttcggt ttgggaccga agccgcgccg 1081 cgcgtcttgt ctgctgcagc atcgttctgt gttgtctctg tctgactgtg tttctgtatt 1141 tgtctgaaaa tatgggcccg ggccagactg ttaccactcc cttaagtttg accttaggtc 1201 actggaaaga tgtcgagcgg atcgctcaca accagtcggt agatgtcaag aagagacgtt 1261 gggttacctt ctgctctgca gaatggccaa cctttaacgt cggatggccg cgagacggca 1321 cctttaaccg agacctcatc acccaggtta agatcaaggt cttttcacct ggcccgcatg 1381 gacacccaga ccaggtcccc tacatcgtga cctgggaagc cttggctttt gacccccctc 1441 cctgggtcaa gccctttgta caccctaagc ctccgcctcc tcttcctcca tccgccccgt 1501 ctctccccct tgaacctcct cgttcgaccc cgcctcgatc ctccctttat ccagccctca 1561 ctccttctct aggcgccccc atatggccat atgagatctt atatggggca cccccgcccc 1621 ttgtaaactt ccctgaccct gacatgacaa gagttactaa cagcccctct ctccaagctc 1681 acttacaggc tctctactta gtccagcacg aagtctggag acctctggcg gcagcctacc 1741 aagaacaact ggaccgaccg gtggtacctc acccttaccg agtcggcgac acagtgtggg 1801 tccgccgaca ccagactaag aacctagaac ctcgctggaa aggaccttac acagtcctgc 1861 tgaccacccc caccgccctc aaagtagacg gcatcgcagc ttggatacac gccgcccacg 1921 tgaaggctgc cgaccccggg ggtggaccat cctctagact gccggatctg ccatcgatat 1981 aaagcttgcc accatggact acaaagacca tgacggtgat tataaagatc atgacatcga 2041 ctacaaggat gacgatgaca agggttcagg ctcgggatcc atgtctgaaa attccagtga 2101 cagtgattca tcttgtggtt ggactgtcat cagtcatgag gggtcagata tagaaatgtt 2161 gaattctgtg acccccactg acagctgtga gcccgcccca gaatgttcat ctttagagca 2221 agaggagctt caagcattgc agatagagca aggagaaagc agccaaaatg gcacagtgct 2281 tatggaagaa actgcttatc cagctttgga ggaaaccagc tcaacaattg aggcagagga 2341 acaaaagata cccgaagaca gtatctatat tggaactgcc agtgatgatt ctgatattgt 2401 tacccttgag ccacctaagt tagaagaaat tggaaatcaa gaagttgtca ttgttgaaga 2461 agcacagagt tcagaagact ttaacatggg ctcttcctct agcagccagt atactttctg 2521 tcagccagaa actgtatttt catctcagcc tagtgacgat gaatcaagta gtgatgaaac 2581 cagtaatcag cccagtcctg cctttagacg acgccgtgct aggaagaaga ccgtttctgc 2641 ttcagaatct gaagaccggc tagttgctga acaagaaact gaaccttcta aggagttgag 2701 taaacgtcag ttcagtagtg gtctcaataa gtgtgttata cttgctttgg tgattgcaat 2761 cagcatggga tttggccatt tctatggcac aattcagatt cagaagcgtc aacagttagt 2821 cagaaagata catgaagatg aattgaatga tatgaaggat tatctttccc agtgtcaaca 2881 ggaacaagaa tcttttatag attataagtc attgaaagaa aatcttgcaa ggtgttggac 2941 acttactgaa gcagagaaga tgtcctttga aactcagaaa acgaaccttg ctacagaaaa 3001 tcagtattta agagtatccc tggagaagga agaaaaagcc ttatcctcat tacaggaaga 3061 gttaaacaaa ctaagagaac agattagaat attggaagat aaagggacaa gtactgaatt 3121 agttaaagaa aatcagaaac ttaagcagca tttggaagag gaaaagcaga aaaaacacag 3181 ctttcttagt caaagggaga ctctgttgac agaagcaaag atgctaaaga gagaactgga 3241 gagagaacga ctagtaacta cggctttaag gggggaactc cagcagttaa gtggtagtca 3301 gttacatggc aagtcagatt ctcccaatgt atatactgaa aaaaaggaaa tagcaatctt 3361 acgggaaaga ctcactgagc tggaacggaa gctaaccttc gaacagcagc gttctgattt 3421 gtgggaaaga ttgtatgttg aggcaaaaga tcaaaatgga aaacaaggaa cagatggaaa 3481 aaagaaaggg ggcagaggaa gccacagggc taaaaataag tcaaaggaaa catttttggg 3541 ttcagttaag gaaacatttg atgccatgaa gaattctacc aaggagtttg taaggcatca 3601 taaagagaaa attaagcagg ctaaagaagc tgtgaaggaa aatctgaaaa aattctcaga 3661 ttcagttaaa tccactttca gacactttaa agataccacc aagaatatct ttgatgaaaa 3721 gggtaataaa agatttggtg ctacaaaaga agcagctgaa aaaccaagaa cagtttttag 3781 tgactattta catccacagt ataaggcacc tacagaaaac catcataata gaggccctac 3841 tatgcaaaat gatggaagga aagaaaagcc agttcacttt aaagaattca gaaaaaatac 3901 aaattcaaag aaatgcagtc ctgggcatga ttgtagagaa aattctcatt ctttcagaaa 3961 ggcttgttct ggtgtatttg attgtgctca acaagagtcc atgagccttt ttaacacagt 4021 ggtgaatcct ataaggatgg atgaatttag acagataatt caaaggtaca tgttaaaaga 4081 actggatact ttttgtcact ggaacgaact tgatcagttc atcaataagt ttttcctaaa 4141 cggtgtcttt atacatgatc agaagctctt cactgacttt gttaatgatg ttaaagatta 4201 tcttagaaac atgaaggaat atgaagtaga taatgatgga gtatttgaga agttggatga 4261 atatatatat agacacttct ttggtcacac tttttcccct ccatatggac ccaggtcggt 4321 ttacataaaa ccgtgtcatt acagtagttt gtaactcgag agcggccgct acgtaaattc 4381 cgcccccccc cctaacgtta ctggccgaag ccgcttggaa taaggccggt gtgcgtttgt 4441 ctatatgtta ttttccacca tattgccgtc ttttggcaat gtgagggccc ggaaacctgg 4501 ccctgtcttc ttgacgagca ttcctagggg tctttcccct ctcgccaaag gaatgcaagg 4561 tctgttgaat gtcgtgaagg aagcagttcc tctggaagtt tcttgaagac aaacaacgtc 4621 tgtagcgacc ctttgcaggc agcggaaccc cccacctggc gacaggtgcc tctgcggcca 4681 aaagccacgt gtataagata cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag 4741 ttggatagtt gtggaaagag tcaaatggct ctcctcaagc gtattcaaca aggggctgaa 4801 ggatgcccag aaggtacccc attgtatggg atctgatctg gggcctcggt gcacatgctt 4861 tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc ccgaaccacg gggacgtggt 4921 tttcctttga aaaacacgat gataatatgg ccacaaccgt cgacatgatt gaacaagatg 4981 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac 5041 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg 5101 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc 5161 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg 5221 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc 5281 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc 5341 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta 5401 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg 5461 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg 5521 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat 5581 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc 5641 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta 5701 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag 5761 tcgaccgccc gccccacgac ccgcagcgcc cgaccgaaag gagcgcacga ccccatggct 5821 cgacgataaa ataaaagatt ttatttagtc tccagaaaaa ggggggaatg aaagacccca 5881 cctgtaggtt tggcaagcta gcttaagtaa cgccattttg caaggcatgg aaaaatacat 5941 aactgagaat agagaagttc agatcaaggt caggaacaga tggaacagct gaatatgggc 6001 caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga acagatggaa 6061 cagctgaata tgggccaaac aggatatctg tggtaagcag ttcctgcccc ggctcagggc 6121 caagaacaga tggtccccag atgcggtcca gccctcagca gtttctagag aaccatcaga 6181 tgtttccagg gtgccccaag gacctgaaat gaccctgtgc cttatttgaa ctaaccaatc 6241 agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg agctcaataa aagagcccac 6301 aacccctcac tcggggcgcc agtcctccga ttgactgagt cgcccgggta cccgtgtatc 6361 caataaaccc tcttgcagtt gcatccgact tgtggtctcg ctgttccttg ggagggtctc 6421 ctctgagtga ttgactaccc gtcagcgggg gtctttcaca tgcagcatgt atcaaaatta 6481 atttggtttt ttttcttaag tatttacatt aaatggccat agttgcatta atgaatcggc 6541 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac 6601 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata 6661 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa 6721 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct 6781 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa 6841 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg 6901 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca 6961 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa 7021 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg 7081 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg 7141 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga 7201 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc 7261 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag 7321 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac 7381 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc 7441 ttcacctaga tccttttgcg gccggccgca aatcaatcta aagtatatat gagtaaactt 7501 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc 7561 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac 7621 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat 7681 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg 7741 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata 7801 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta 7861 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt 7921 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag 7981 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa 8041 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc 8101 gaccgagttg ctcttgcccg gcgtcaacac gggataatac cgcgccacat agcagaactt 8161 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc 8221 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta 8281 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa 8341 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca 8401 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac 8461 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta 8521 ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt cttcactata 8581 ggagcttggc ccattgcata cgttgtatcc atatcataat atgtacattt atattggctc 8641 atgtccaaca ttaccgccat gtt //