LOCUS pCMV6-AC-IRES-GFP 7272 bp ds-DNA circular SYN 14-FEB-2014 DEFINITION PrecisionShuttle(TM) mammalian expression vector with a TurboGFP marker, for fusing an ORF to C-terminal c-Myc and FLAG(R) tags. ACCESSION . VERSION . KEYWORDS pCMV6-AC-IRES-GFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7272) AUTHORS OriGene TITLE Direct Submission JOURNAL Exported Saturday, June 13, 2020 from SnapGene Viewer 3.1.4 http://www.snapgene.com COMMENT Inserts are typically cloned between the SgfI (AsiSI) and MluI sites. FEATURES Location/Qualifiers source 1..7272 /organism= synthetic DNA construct /lab_host= Mammalian Cells /mol_type= other DNA enhancer 343..722 /note= CMV enhancer /note= human cytomegalovirus immediate early enhancer promoter 723..926 /note= CMV promoter /note= human cytomegalovirus (CMV) immediate early promoter promoter 952..970 /note= T7 promoter /note= promoter for bacteriophage T7 RNA polymerase misc_feature 979..1087 /note= MCS /note= multiple cloning site CDS 1085..1114 /codon_start=1 /product= Myc (human c-Myc oncogene) epitope tag /note= Myc /translation= EQKLISEEDL CDS 1133..1156 /codon_start=1 /product= FLAG(R) epitope tag, followed by an enterokinase cleavage site /note= FLAG /translation= DYKDDDDK misc_feature 1184..1770 /note= IRES2 /note= internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV) CDS 1759..1761 /codon_start=1 /product= start codon /note= ATG /translation= M CDS 1771..2469 /codon_start=1 /product= improved green fluorescent protein from Pontellina plumata (Evdokimov et al., 2006) /note= TurboGFP /note= mammalian codon-optimized /translation= MESDESGLPAMEIECRITGTLNGVEFELVGGGEGTPEQGRMTNKM KSTKGALTFSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVL HVSFSYRYEAGRVIGDFKVMGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNDLDGSFTR TFSLRDGGYYSSVVDSHMHFKSAIHPSILQNGGPMFAFRRVEEDHSNTELGIVEYQHAF KTPDADAGEE polyA_signal 2505..3127 /note= hGH poly(A) signal /note= human growth hormone polyadenylation signal promoter 3156..3485 /note= SV40 promoter /note= SV40 enhancer and early promoter rep_origin 3336..3471 /note= SV40 ori /note= SV40 origin of replication CDS 3552..4346 /codon_start=1 /gene= aph(3 )-II (or nptII) /product= aminoglycoside phosphotransferase from Tn5 /note= NeoR/KanR /note= confers resistance to neomycin, kanamycin, and G418 (Geneticin(R)) /translation= MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF polyA_signal 4520..4641 /note= SV40 poly(A) signal /note= SV40 polyadenylation signal rep_origin complement(5092..5679) /direction=LEFT /note= ori /note= high-copy-number ColE1/pMB1/pBR322/pUC origin of replication CDS complement(5850..6710) /codon_start=1 /gene= bla /product= beta-lactamase /note= AmpR /note= confers resistance to ampicillin, carbenicillin, and related antibiotics /translation= MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW promoter complement(6711..6815) /gene= bla /note= AmpR promoter rep_origin 6842..25 /direction=RIGHT /note= f1 ori /note= f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis ORIGIN 1 aacaaaatat taacgcttac aatttccatt cgccattcag gctgcgcaac tgttgggaag 61 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa 121 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca 181 gtgccaagct gatctataca ttgaatcaat attggcaatt agccatatta gtcattggtt 241 atatagcata aatcaatatt ggctattggc cattgcatac gttgtatcta tatcataata 301 tgtacattta tattggctca tgtccaatat gaccgccatg ttgacattga ttattgacta 361 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg 421 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga 481 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat 541 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa 601 gtccgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca 661 tgaccttacg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca 721 tggtgatgcg gttttggcag tacaccaatg ggcgtggata gcggtttgac tcacggggat 781 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg 841 actttccaaa atgtcgtaat aaccccgccc cgttgacgca aatgggcggt aggcgtgtac 901 ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagaatttt gtaatacgac 961 tcactatagg gcggccggga attcgtcgac tggatccggt accgaggaga tctgccgccg 1021 cgatcgccgg cgcgccagat ctcaagctta actagctagc ggaccgacgc gtacgcggcc 1081 gctcgagcag aaactcatct cagaagagga tctggcagca aatgatatcc tggattacaa 1141 ggatgacgac gataaggttt aaacgtagcg gccgcaaatt ccgcccctct ccctcccccc 1201 cccctaacgt tactggccga agccgcttgg aataaggccg gtgtgcgttt gtctatatgt 1261 tattttccac catattgccg tcttttggca atgtgagggc ccggaaacct ggccctgtct 1321 tcttgacgag cattcctagg ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga 1381 atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag acaaacaacg tctgtagcga 1441 ccctttgcag gcagcggaac cccccacctg gcgacaggtg cctctgcggc caaaagccac 1501 gtgtataaga tacacctgca aaggcggcac aaccccagtg ccacgttgtg agttggatag 1561 ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg aaggatgccc 1621 agaaggtacc ccattgtatg ggatctgatc tggggcctcg gtgcacatgc tttacatgtg 1681 tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg gttttccttt 1741 gaaaaacacg atgataatat ggccacaacc atggagagcg acgagagcgg cctgcccgcc 1801 atggagatcg agtgccgcat caccggcacc ctgaacggcg tggagttcga gctggtgggc 1861 ggcggagagg gcacccccga gcagggccgc atgaccaaca agatgaagag caccaaaggc 1921 gccctgacct tcagccccta cctgctgagc cacgtgatgg gctacggctt ctaccacttc 1981 ggcacctacc ccagcggcta cgagaacccc ttcctgcacg ccatcaacaa cggcggctac 2041 accaacaccc gcatcgagaa gtacgaggac ggcggcgtgc tgcacgtgag cttcagctac 2101 cgctacgagg ccggccgcgt gatcggcgac ttcaaggtga tgggcaccgg cttccccgag 2161 gacagcgtga tcttcaccga caagatcatc cgcagcaacg ccaccgtgga gcacctgcac 2221 cccatgggcg ataacgatct ggatggcagc ttcacccgca ccttcagcct gcgcgacggc 2281 ggctactaca gctccgtggt ggacagccac atgcacttca agagcgccat ccaccccagc 2341 atcctgcaga acgggggccc catgttcgcc ttccgccgcg tggaggagga tcacagcaac 2401 accgagctgg gcatcgtgga gtaccagcac gccttcaaga ccccggatgc agatgccggt 2461 gaagaataat agccgcggtc atagctgttt cctgaacaga tcccgggtgg catccctgtg 2521 acccctcccc agtgcctctc ctggccctgg aagttgccac tccagtgccc accagccttg 2581 tcctaataaa attaagttgc atcattttgt ctgactaggt gtccttctat aatattatgg 2641 ggtggagggg ggtggtatgg agcaaggggc aagttgggaa gacaacctgt agggcctgcg 2701 gggtctattg ggaaccaagc tggagtgcag tggcacaatc ttggctcact gcaatctccg 2761 cctcctgggt tcaagcgatt ctcctgcctc agcctcccga gttgttggga ttccaggcat 2821 gcatgaccag gctcagctaa tttttgtttt tttggtagag acggggtttc accatattgg 2881 ccaggctggt ctccaactcc taatctcagg tgatctaccc accttggcct cccaaattgc 2941 tgggattaca ggcgtgaacc actgctccct tccctgtcct tctgatttta aaataactat 3001 accagcagga ggacgtccag acacagcata ggctacctgg ccatgcccaa ccggtgggac 3061 atttgagttg cttgcttggc actgtcctct catgcgttgg gtccactcag tagatgcctg 3121 ttgaattggg tacgcggcca gcttggctgt ggaatgtgtg tcagttaggg tgtggaaagt 3181 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca 3241 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt 3301 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt 3361 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg 3421 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt 3481 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag 3541 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg 3601 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt 3661 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc 3721 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt 3781 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag 3841 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg 3901 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag 3961 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg 4021 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc 4081 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca 4141 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggccgacc 4201 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg 4261 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct 4321 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc 4381 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg 4441 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct 4501 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa 4561 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc 4621 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc 4681 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa 4741 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac 4801 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca 4861 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc 4921 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc 4981 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc 5041 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag 5101 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc 5161 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt 5221 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct 5281 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg 5341 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct 5401 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat 5461 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg 5521 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa 5581 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gttttttgtt 5641 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct 5701 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta 5761 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa 5821 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc 5881 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact 5941 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc 6001 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt 6061 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta 6121 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg 6181 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt 6241 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc 6301 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt 6361 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc 6421 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc 6481 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa 6541 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac 6601 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa 6661 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt 6721 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa 6781 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct 6841 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc 6901 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc 6961 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt 7021 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg 7081 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt 7141 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta 7201 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt 7261 aacgcgaatt tt