LOCUS       pCMV6-AC-IRES-GF        7272 bp DNA     circular SYN 01-JAN-1980
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
REFERENCE   2  (bases 1 to 7272)
  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..7272
                     /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    2505..3127
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     promoter        3156..3485
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3552..4343
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     polyA_signal    4520..4653
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4690..4706)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4714..4730)
                     /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(4738..4768)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4783..4804)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5092..5679)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5853..6710)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(6711..6815)
                     /label=AmpR promoter
     rep_origin      6842..7272
                     /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 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
//
