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
//
