LOCUS       pcDNA3_N3FLAG_MY        6474 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..912,1999..2022)
                     /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(2026..2044)
                     /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    2070..2294
                     /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      2340..2768
                     /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
     promoter        2782..3112
                     /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      2963..3098
                     /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             3179..3973
                     /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    4147..4268
                     /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
     misc_feature    1018..1995
                     /label=MYB-(1-327)
                     /ApEinfo_fwdcolor=#ff6666
                     /ApEinfo_revcolor=green
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     primer_bind     complement(4317..4333)
                     /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(4719..5307)
                     /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(5478..6338)
                     /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(6339..6443)
                     /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 GCAGGAGCCT
     1621 TCCAAAGCCA GCCAGACGCC AGTGGCCACG AGCTTCCAGA AGAACAATCA TTTGATGGGG
     1681 TTTGGGCATG CCTCACCTCC ATCTCAGCTC TCTCCAAGTG GCCAGTCCTC CGTCAACAGC
     1741 GAATATCCCT ATTACCACAT CGCCGAAGCA CAAAACATCT CCAGTCACGT TCCCTATCCT
     1801 GTCGCATTGC ATGTTAATAT AGTCAACGTC CCTCAGCCGG CTGCGGCAGC CATCCAGAGA
     1861 CACTATAACG ACGAAGACCC TGAGAAGGAA AAGCGAATAA AGGAGCTGGA GTTGCTCCTG
     1921 ATGTCAACAG AGAACGAGCT GAAGGGACAG CAGGCATTAC CAACACAGAA CCACACTTGC
     1981 AGCTACCCCG GGTGGtgact cgagcatgca tctagagggc cctattctat agtgtcacct
     2041 aaatgctaga gctcgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     2101 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     2161 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     2221 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
     2281 ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca
     2341 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     2401 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     2461 gttcgccggc tttccccgtc aagctctaaa tcggggcatc cctttagggt tccgatttag
     2521 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     2581 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     2641 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     2701 agggattttg gggatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     2761 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     2821 ggcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     2881 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     2941 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     3001 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga
     3061 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc
     3121 gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat
     3181 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     3241 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     3301 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     3361 ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     3421 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     3481 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     3541 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     3601 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     3661 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg
     3721 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     3781 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     3841 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     3901 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     3961 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
     4021 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     4081 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     4141 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     4201 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     4261 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca
     4321 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     4381 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     4441 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     4501 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     4561 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     4621 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     4681 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     4741 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     4801 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     4861 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcaatgctca
     4921 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     4981 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     5041 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     5101 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     5161 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     5221 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     5281 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     5341 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     5401 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     5461 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     5521 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     5581 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     5641 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     5701 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     5761 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     5821 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     5881 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     5941 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     6001 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     6061 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     6121 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     6181 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     6241 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     6301 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     6361 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     6421 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtc
//
