LOCUS       pcDNA3_N3FLAG_MY        6093 bp ds-DNA     circular     18-DEC-2017
DEFINITION  Mammalian expression vector with the CMV promoter and a
            neomycin-resistance marker.
ACCESSION   .
VERSION     .
KEYWORDS    pcDNA3
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5446)
  AUTHORS   Invitrogen (Life Technologies)
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Dec 18, 2017 from SnapGene Viewer 3.3.4
            http://www.snapgene.com
COMMENT     The cloned gene must include a Kozak sequence and start codon.
COMMENT     
COMMENT     ApEinfo:methylated:1
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label=CMV enhancer
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label=CMV promoter
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label=T7 promoter
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     misc_feature    916..918
                     /label=3*Flag
                     /ApEinfo_fwdcolor=#cc66ff
                     /ApEinfo_revcolor=green
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     misc_feature    join(889..912,1618..1641)
                     /note="multiple cloning site"
                     /label=MCS
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     misc_feature    940..1005
                     /label=3*Flag(1)
                     /ApEinfo_label=3*Flag
                     /ApEinfo_fwdcolor=#cc66ff
                     /ApEinfo_revcolor=green
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     promoter        complement(1645..1663)
                     /note="promoter for bacteriophage SP6 RNA polymerase"
                     /label=SP6 promoter
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     misc_feature    919..939
                     /label=NLS
                     /ApEinfo_fwdcolor=#00ff00
                     /ApEinfo_revcolor=green
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     polyA_signal    1689..1913
                     /note="bovine growth hormone polyadenylation signal"
                     /label=bGH poly(A) signal
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     misc_feature    1012..1017
                     /label=EcoRI
                     /ApEinfo_fwdcolor=#ff6fcf
                     /ApEinfo_revcolor=green
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     rep_origin      1959..2387
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label=f1 ori
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     misc_feature    1018..1617
                     /label=MYB-DBD(aa 1-200)
                     /ApEinfo_fwdcolor=#ff6666
                     /ApEinfo_revcolor=green
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     promoter        2401..2731
                     /note="SV40 enhancer and early promoter"
                     /label=SV40 promoter
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     rep_origin      2582..2717
                     /note="SV40 origin of replication"
                     /label=SV40 ori
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     CDS             2798..3592
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin)"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR
                     P
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLL
                     S
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEH
                     Q
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDI
                     A LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
                     /label=NeoR/KanR
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     polyA_signal    3766..3887
                     /note="SV40 polyadenylation signal"
                     /label=SV40 poly(A) signal
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     primer_bind     complement(3936..3952)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label=M13 rev
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     rep_origin      complement(4338..4926)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label=ori
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     CDS             complement(5097..5957)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="confers resistance to ampicillin, carbenicillin,
                     and related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     I
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     S
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     W
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     A
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     S LIKHW"
                     /label=AmpR
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     promoter        complement(5958..6062)
                     /gene="bla"
                     /label=AmpR promoter
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 gagctcggat ccGCCATGCC AAAGAAGAAG CGTAAGGTAG ATTACAAAGA CCATGACGGT
      961 GATTATAAAG ATCATGATAT CGATTACAAG GATGACGATG ACAAGGGAGG CgaattcGCC
     1021 CGGAGACCCC GACACAGCAT CTACAGTAGC GATGAAGATG ATGAAGACAT TGAGATGTGT
     1081 GACCATGACT ACGATGGGCT GCTGCCCAAA TCTGGAAAGC GTCACTTGGG GAAAACTAGG
     1141 TGGACAAGGG AAGAGGATGA GAAGCTGAAG AAGCTGGTGG AACAGAACGG AACAGACGAC
     1201 TGGAAAGTCA TTGCCAATTA TCTGCCCAAC CGGACAGATG TGCAGTGCCA ACACCGGTGG
     1261 CAGAAAGTGC TGAACCCTGA ACTCATCAAA GGTCCCTGGA CCAAAGAAGA AGATCAGAGA
     1321 GTCATAGAGC TTGTCCAGAA ATATGGTCCG AAGCGTTGGT CTGTTATTGC CAAGCACTTA
     1381 AAAGGGAGAA TTGGAAAGCA GTGTCGGGAG AGGTGGCACA ACCATTTGAA TCCAGAAGTT
     1441 AAGAAAACCT CCTGGACAGA AGAGGAGGAC AGAATCATTT ACCAGGCACA CAAGCGTCTG
     1501 GGGAACAGAT GGGCAGAGAT CGCAAAGCTG CTGCCCGGAC GGACTGATAA TGCTATCAAG
     1561 AACCACTGGA ATTCCACCAT GCGTCGCAAG GTGGAACAGG AAGGCTACCT GCAGTGActc
     1621 gagcatgcat ctagagggcc ctattctata gtgtcaccta aatgctagag ctcgctgatc
     1681 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     1741 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     1801 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     1861 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga
     1921 ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
     1981 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     2041 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     2101 agctctaaat cggggcatcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     2161 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     2221 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     2281 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgg ggatttcggc
     2341 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
     2401 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag gcaggcagaa gtatgcaaag
     2461 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     2521 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     2581 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     2641 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg
     2701 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
     2761 cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     2821 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     2881 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     2941 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
     3001 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     3061 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     3121 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     3181 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     3241 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     3301 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
     3361 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     3421 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     3481 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     3541 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     3601 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     3661 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     3721 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
     3781 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     3841 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
     3901 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     3961 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     4021 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     4081 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     4141 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     4201 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     4261 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     4321 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     4381 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     4441 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     4501 tttctccctt cgggaagcgt ggcgctttct caatgctcac gctgtaggta tctcagttcg
     4561 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     4621 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     4681 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     4741 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
     4801 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     4861 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     4921 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     4981 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     5041 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     5101 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     5161 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     5221 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     5281 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     5341 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     5401 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     5461 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     5521 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     5581 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     5641 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     5701 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     5761 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     5821 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     5881 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     5941 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     6001 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     6061 cgcacatttc cccgaaaagt gccacctgac gtc  
//
