LOCUS       pCMV6-AN-GFP-AC-        6616 bp DNA     circular SYN 01-JAN-1980
DEFINITION  PrecisionShuttle(TM) mammalian expression vector for fusing an ORF 
            to an N-terminal TurboGFP tag and a C-terminal 6xHis tag.
ACCESSION   .
VERSION     .
KEYWORDS    pCMV6-AN-GFP-AC-His
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6616)
  AUTHORS   OriGene
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 6616)
  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..6616
                     /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"
     CDS             1020..1715
                     /label=TurboGFP
                     /note="improved green fluorescent protein from Pontellina 
                     plumata (Evdokimov et al., 2006)"
     misc_feature    1719..1786
                     /label=MCS
                     /note="MCS"
                     /note="multiple cloning site"
     CDS             1787..1804
                     /label=6xHis
                     /note="6xHis affinity tag"
     polyA_signal    1851..2473
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     promoter        2502..2831
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             2898..3689
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     polyA_signal    3866..3999
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4036..4052)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4060..4076)
                     /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(4084..4114)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4129..4150)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4438..5023)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5197..6054)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(6055..6159)
                     /label=AmpR promoter
     rep_origin      6186..6616
                     /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 tctgccacca
     1021 tggagagcga cgagagcggc ctgcccgcca tggagatcga gtgccgcatc accggcaccc
     1081 tgaacggcgt ggagttcgag ctggtgggcg gcggagaggg cacccccgag cagggccgca
     1141 tgaccaacaa gatgaagagc accaaaggcg ccctgacctt cagcccctac ctgctgagcc
     1201 acgtgatggg ctacggcttc taccacttcg gcacctaccc cagcggctac gagaacccct
     1261 tcctgcacgc catcaacaac ggcggctaca ccaacacccg catcgagaag tacgaggacg
     1321 gcggcgtgct gcacgtgagc ttcagctacc gctacgaggc cggccgcgtg atcggcgact
     1381 tcaaggtgat gggcaccggc ttccccgagg acagcgtgat cttcaccgac aagatcatcc
     1441 gcagcaacgc caccgtggag cacctgcacc ccatgggcga taacgatctg gatggcagct
     1501 tcacccgcac cttcagcctg cgcgacggcg gctactacag ctccgtggtg gacagccaca
     1561 tgcacttcaa gagcgccatc caccccagca tcctgcagaa cgggggcccc atgttcgcct
     1621 tccgccgcgt ggaggaggat cacagcaaca ccgagctggg catcgtggag taccagcacg
     1681 ccttcaagac cccggatgca gatgccggtg aagaaagagc gatcgccggc gcgccagatc
     1741 tcaagcttaa ctagctagcg gaccgacgcg tacgcggccg ctcgagcatc accatcacca
     1801 tcacgtttaa acggccggcc gcggtcatag ctgtttcctg aacagatccc gggtggcatc
     1861 cctgtgaccc ctccccagtg cctctcctgg ccctggaagt tgccactcca gtgcccacca
     1921 gccttgtcct aataaaatta agttgcatca ttttgtctga ctaggtgtcc ttctataata
     1981 ttatggggtg gaggggggtg gtatggagca aggggcaagt tgggaagaca acctgtaggg
     2041 cctgcggggt ctattgggaa ccaagctgga gtgcagtggc acaatcttgg ctcactgcaa
     2101 tctccgcctc ctgggttcaa gcgattctcc tgcctcagcc tcccgagttg ttgggattcc
     2161 aggcatgcat gaccaggctc agctaatttt tgtttttttg gtagagacgg ggtttcacca
     2221 tattggccag gctggtctcc aactcctaat ctcaggtgat ctacccacct tggcctccca
     2281 aattgctggg attacaggcg tgaaccactg ctcccttccc tgtccttctg attttaaaat
     2341 aactatacca gcaggaggac gtccagacac agcataggct acctggccat gcccaaccgg
     2401 tgggacattt gagttgcttg cttggcactg tcctctcatg cgttgggtcc actcagtaga
     2461 tgcctgttga attgggtacg cggccagctt ggctgtggaa tgtgtgtcag ttagggtgtg
     2521 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     2581 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     2641 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     2701 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     2761 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     2821 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag
     2881 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     2941 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     3001 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     3061 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     3121 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     3181 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     3241 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     3301 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     3361 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     3421 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     3481 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     3541 ccgaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     3601 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     3661 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     3721 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     3781 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     3841 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     3901 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     3961 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     4021 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     4081 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     4141 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     4201 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     4261 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     4321 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     4381 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     4441 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     4501 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     4561 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     4621 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     4681 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     4741 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     4801 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     4861 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     4921 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtttttt
     4981 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     5041 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     5101 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     5161 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     5221 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     5281 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     5341 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     5401 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     5461 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     5521 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     5581 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     5641 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     5701 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     5761 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     5821 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     5881 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     5941 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     6001 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     6061 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     6121 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     6181 acctgacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     6241 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     6301 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     6361 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     6421 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     6481 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     6541 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     6601 atttaacgcg aatttt
//
