LOCUS       pcDNA6.2_EmGFP-B        8198 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Vivid Colors(TM) Gateway(R) destination vector for expressing 
            proteins in mammalian cells, with an EmGFP-Bsd fusion as a 
            transfection marker.
ACCESSION   .
VERSION     .
KEYWORDS    pcDNA6.2 EmGFP-Bsd V5-DEST.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8198)
  AUTHORS   Invitrogen (Life Technologies)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 8198)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8198
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    911..1035
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(1467..1769)
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     CDS             complement(2117..2773)
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     promoter        complement(2774..2876)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     protein_bind    complement(3054..3178)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             3204..3245
                     /label=V5 tag
                     /note="epitope tag from simian virus 5"
     polyA_signal    3329..3377
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3579..4007
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4022..4521
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     promoter        4535..4582
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter"
     CDS             4601..5317
                     /label=EmGFP
                     /note="Emerald GFP"
     CDS             5318..5713
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=Aspergillus terreus BSD
                     /note="BSD"
                     /note="confers resistance to blasticidin"
                     /translation="AKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTGV
                     NVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGIK
                     AIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    5871..6004
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6041..6057)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(6065..6081)
                     /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(6089..6119)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(6134..6155)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6443..7031)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7205..8062)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(8063..8167)
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct 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 atgggtggag 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 gctggctagt
      901 taagctatca acaagtttgt acaaaaaagc tgaacgagaa acgtaaaatg atataaatat
      961 caatatatta aattagattt tgcataaaaa acagactaca taatactgta aaacacaaca
     1021 tatccagtca ctatgaatca actacttaga tggtattagt gacctgtagt cgaccgacag
     1081 ccttccaaat gttcttcggg tgatgctgcc aacttagtcg accgacagcc ttccaaatgt
     1141 tcttctcaaa cggaatcgtc gtatccagcc tactcgctat tgtcctcaat gccgtattaa
     1201 atcataaaaa gaaataagaa aaagaggtgc gagcctcttt tttgtgtgac aaaataaaaa
     1261 catctaccta ttcatatacg ctagtgtcat agtcctgaaa atcatctgca tcaagaacaa
     1321 tttcacaact cttatacttt tctcttacaa gtcgttcggc ttcatctgga ttttcagcct
     1381 ctatacttac taaacgtgat aaagtttctg taatttctac tgtatcgacc tgcagactgg
     1441 ctgtgtataa gggagcctga catttatatt ccccagaaca tcaggttaat ggcgtttttg
     1501 atgtcatttt cgcggtggct gagatcagcc acttcttccc cgataacgga gaccggcaca
     1561 ctggccatat cggtggtcat catgcgccag ctttcatccc cgatatgcac caccgggtaa
     1621 agttcacggg agactttatc tgacagcaga cgtgcactgg ccagggggat caccatccgt
     1681 cgcccgggcg tgtcaataat atcactctgt acatccacaa acagacgata acggctctct
     1741 cttttatagg tgtaaacctt aaactgcatt tcaccagtcc ctgttctcgt cagcaaaaga
     1801 gccgttcatt tcaataaacc gggcgacctc agccatccct tcctgatttt ccgctttcca
     1861 gcgttcggca cgcagacgac gggcttcatt ctgcatggtt gtgcttacca gaccggagat
     1921 attgacatca tatatgcctt gagcaactga tagctgtcgc tgtcaactgt cactgtaata
     1981 cgctgcttca tagcacacct ctttttgaca tacttcgggt atacatatca gtatatattc
     2041 ttataccgca aaaatcagcg cgcaaatacg catactgtta tctggctttt agtaagccgg
     2101 atccacgcgg cgtttacgcc ccgccctgcc actcatcgca gtactgttgt aattcattaa
     2161 gcattctgcc gacatggaag ccatcacaaa cggcatgatg aacctgaatc gccagcggca
     2221 tcagcacctt gtcgccttgc gtataatatt tgcccatggt gaaaacgggg gcgaagaagt
     2281 tgtccatatt ggccacgttt aaatcaaaac tggtgaaact cacccaggga ttggctgaga
     2341 cgaaaaacat attctcaata aaccctttag ggaaataggc caggttttca ccgtaacacg
     2401 ccacatcttg cgaatatatg tgtagaaact gccggaaatc gtcgtggtat tcactccaga
     2461 gcgatgaaaa cgtttcagtt tgctcatgga aaacggtgta acaagggtga acactatccc
     2521 atatcaccag ctcaccgtct ttcattgcca tacggaattc cggatgagca ttcatcaggc
     2581 gggcaagaat gtgaataaag gccggataaa acttgtgctt atttttcttt acggtcttta
     2641 aaaaggccgt aatatccagc tgaacggtct ggttataggt acattgagca actgactgaa
     2701 atgcctcaaa atgttcttta cgatgccatt gggatatatc aacggtggta tatccagtga
     2761 tttttttctc cattttagct tccttagctc ctgaaaatct cgataactca aaaaatacgc
     2821 ccggtagtga tcttatttca ttatggtgaa agttggaacc tcttacgtgc cgatcaacgt
     2881 ctcattttcg ccaaaagttg gcccagggct tcccggtatc aacagggaca ccaggattta
     2941 tttattctgc gaagtgatct tccgtcacag gtatttattc ggcgcaaagt gcgtcgggtg
     3001 atgctgccaa cttagtcgac tacaggtcac taataccatc taagtagttg attcatagtg
     3061 actggatatg ttgtgtttta cagtattatg tagtctgttt tttatgcaaa atctaattta
     3121 atatattgat atttatatca ttttacgttt ctcgttcagc tttcttgtac aaagtggttg
     3181 atctagaggg cccgcggttc gaaggtaagc ctatccctaa ccctctcctc ggtctcgatt
     3241 ctacgcgtac cggttagtaa tgagtttaaa cgggggaggc taactgaaac acggaaggag
     3301 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggg
     3361 tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac
     3421 cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc
     3481 cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat
     3541 agcagatctg cgcagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
     3601 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     3661 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     3721 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     3781 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     3841 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     3901 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     3961 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattatc ctcgagctac
     4021 cgggtagggg aggcgctttt cccaaggcag tctggagcat gcgctttagc agccccgctg
     4081 ggcacttggc gctacacaag tggcctctgg cctcgcacac attccacatc caccggtagg
     4141 cgccaaccgg ctccgttctt tggtggcccc ttcgcgccac cttctactcc tcccctagtc
     4201 aggaagttcc cccccgcccc gcagctcgcg tcgtgcagga cgtgacaaat ggaagtagca
     4261 cgtctcacta gtctcgtgca gatggacagc accgctgagc aatggaagcg ggtaggcctt
     4321 tggggcagcg gccaatagca gctttgctcc ttcgctttct gggctcagag gctgggaagg
     4381 ggtgggtccg ggggcgggct caggggcggg ctcaggggcg gggcgggcgc ccgaaggtcc
     4441 tccggaggcc cggcattctg cacgcttcaa aagcgcacgt ctgccgcgct gttctcctct
     4501 tcctcatctc cgggcctttc gactctagac acgtgttgac aattaatcat cggcatagta
     4561 tatcggcata gtataatacg acaaggtgag gaactaaacc atggtgagca agggcgagga
     4621 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
     4681 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt
     4741 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccttcaccta
     4801 cggcgtgcag tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     4861 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
     4921 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     4981 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
     5041 cagccacaag gtctatatca ccgccgacaa gcagaagaac ggcatcaagg tgaacttcaa
     5101 gacccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
     5161 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc
     5221 cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
     5281 cgccgggatc actctcggca tggacgagct gtacaaggcc aagcctttgt ctcaagaaga
     5341 atccaccctc attgaaagag caacggctac aatcaacagc atccccatct ctgaagacta
     5401 cagcgtcgcc agcgcagctc tctctagcga cggccgcatc ttcactggtg tcaatgtata
     5461 tcattttact gggggacctt gtgcagaact cgtggtgctg ggcactgctg ctgctgcggc
     5521 agctggcaac ctgacttgta tcgtcgcgat cggaaatgag aacaggggca tcttgagccc
     5581 ctgcggacgg tgccgacagg tgcttctcga tctgcatcct gggatcaaag ccatagtgaa
     5641 ggacagtgat ggacagccga cggcagttgg gattcgtgaa ttgctgccct ctggttatgt
     5701 gtgggagggc taagcacttc gtggccgagg agcaggactg acacgtgcta cgagatttcg
     5761 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     5821 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
     5881 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     5941 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     6001 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     6061 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     6121 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     6181 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     6241 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     6301 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     6361 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     6421 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     6481 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     6541 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     6601 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     6661 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     6721 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     6781 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     6841 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     6901 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     6961 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     7021 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     7081 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     7141 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     7201 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     7261 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     7321 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     7381 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     7441 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     7501 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     7561 tcagcttcgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     7621 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     7681 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     7741 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     7801 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     7861 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     7921 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     7981 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     8041 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     8101 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     8161 ttccgcgcac atttccccga aaagtgccac ctgacgtc
//
