LOCUS pCMV6-AC-IRES-GF 7072 bp DNA circular SYN 01-JAN-1980 DEFINITION PrecisionShuttle(TM) mammalian expression vector with TurboGFP and puromycin resistance markers, for fusing an ORF to C-terminal c-Myc and FLAG(R) tags. ACCESSION . VERSION . KEYWORDS pCMV6-AC-IRES-GFP-Puro SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7072) AUTHORS OriGene TITLE Direct Submission REFERENCE 2 (bases 1 to 7072) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article" COMMENT Inserts are typically cloned between the SgfI (AsiSI) and MluI sites. FEATURES Location/Qualifiers source 1..7072 /mol_type="other DNA" /organism="synthetic DNA construct" primer_bind 166..182 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" enhancer 343..722 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 723..926 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" promoter 952..970 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" misc_feature 979..1087 /label=MCS /note="MCS" /note="multiple cloning site" CDS 1085..1114 /label=Myc /note="Myc (human c-Myc proto-oncogene) epitope tag" CDS 1133..1156 /label=FLAG /note="FLAG(R) epitope tag, followed by an enterokinase cleavage site" misc_feature 1184..1770 /label=IRES2 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" CDS 1771..2466 /label=TurboGFP /note="improved green fluorescent protein from Pontellina plumata (Evdokimov et al., 2006)" polyA_signal 2503..3125 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" promoter 3154..3483 /label=SV40 promoter /note="SV40 enhancer and early promoter" CDS 3541..4137 /label=PuroR /note="puromycin N-acetyltransferase" polyA_signal 4319..4452 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4489..4505) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind complement(4513..4529) /label=lac operator /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-beta-D-thiogalactopyranoside (IPTG)." promoter complement(4537..4567) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind complement(4582..4603) /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(4891..5479) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(5653..6510) /label=AmpR /note="beta-lactamase" promoter complement(6511..6615) /label=AmpR promoter rep_origin 6642..7072 /label=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 agccgctcat agctgtttcc tgaacagatc ccgggtggca tccctgtgac 2521 ccctccccag tgcctctcct ggccctggaa gttgccactc cagtgcccac cagccttgtc 2581 ctaataaaat taagttgcat cattttgtct gactaggtgt ccttctataa tattatgggg 2641 tggagggggg tggtatggag caaggggcaa gttgggaaga caacctgtag ggcctgcggg 2701 gtctattggg aaccaagctg gagtgcagtg gcacaatctt ggctcactgc aatctccgcc 2761 tcctgggttc aagcgattct cctgcctcag cctcccgagt tgttgggatt ccaggcatgc 2821 atgaccaggc tcagctaatt tttgtttttt tggtagagac ggggtttcac catattggcc 2881 aggctggtct ccaactccta atctcaggtg atctacccac cttggcctcc caaattgctg 2941 ggattacagg cgtgaaccac tgctcccttc cctgtccttc tgattttaaa ataactatac 3001 cagcaggagg acgtccagac acagcatagg ctacctggcc atgcccaacc ggtgggacat 3061 ttgagttgct tgcttggcac tgtcctctca tgcgttgggt ccactcagta gatgcctgtt 3121 gaattgggta cgcggccagc ttggctgtgg aatgtgtgtc agttagggtg tggaaagtcc 3181 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg 3241 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag 3301 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc 3361 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc 3421 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc 3481 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac acgtacgacc 3541 atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc ccgggcagta 3601 cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccagac 3661 cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac 3721 atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag 3781 agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt 3841 tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag 3901 cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc 3961 agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg 4021 gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc 4081 gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga 4141 ctgcagtggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg 4201 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga 4261 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa 4321 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa 4381 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta 4441 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt 4501 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa 4561 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact 4621 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc 4681 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg 4741 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc 4801 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag 4861 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca 4921 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca 4981 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg 5041 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag 5101 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt 5161 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca 5221 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg 5281 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt 5341 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc 5401 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg 5461 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg 5521 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta 5581 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg 5641 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg 5701 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc 5761 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc 5821 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc 5881 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag 5941 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat 6001 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg 6061 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt 6121 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag 6181 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg 6241 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt 6301 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct 6361 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac 6421 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat 6481 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat 6541 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca 6601 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgcgccct gtagcggcgc 6661 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct 6721 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg 6781 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga 6841 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt 6901 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg 6961 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc 7021 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt tt //