LOCUS       40924_10366        8577 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector pCeMM-NTAP(GS)-Gw, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8577)
  AUTHORS   Burckstummer T, Bennett KL, Preradovic A, Schutze G, Hantschel O, 
            Superti-Furga G, Bauch A.
  TITLE     An efficient tandem affinity purification procedure for interaction 
            proteomics in mammalian cells
  JOURNAL   Nat. Methods 3 (12), 1013-1019 (2006)
  PUBMED    17060908
REFERENCE   2  (bases 1 to 8577)
  AUTHORS   Burckstummer T, Bennett KL, Preradovic A, Schutze G, Hantschel O, 
            Bauch A, Superti-Furga G.
  TITLE     Direct Submission
  JOURNAL   Submitted (27-NOV-2006) Director's Laboratory, Research Center for 
            Molecular Medicine, Lazarettgasse 19/3, Vienna 1090, Austria
REFERENCE   3  (bases 1 to 8577)
  AUTHORS   Burckstummer T, Bennett KL, Preradovic A, Schutze G, Hantschel O, 
            Bauch A, Superti-Furga G.
  TITLE     Direct Submission
  JOURNAL   Submitted (14-DEC-2006) Director's Laboratory, Research Center for 
            Molecular Medicine, Lazarettgasse 19/3, Vienna 1090, Austria
REFERENCE   4  (bases 1 to 8577)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 8577)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat. 
            Methods"; date: "2006"; volume: "3"; issue: "12"; pages: "1013-1019"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (27-NOV-2006) Director's Laboratory, Research Center for Molecular 
            Medicine, Lazarettgasse 19/3, Vienna 1090, Austria"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Submitted 
            (14-DEC-2006) Director's Laboratory, Research Center for Molecular 
            Medicine, Lazarettgasse 19/3, Vienna 1090, Austria"
COMMENT     SGRef: number: 4; type: "Journal Article"
COMMENT     On Dec 14, 2006 this sequence version replaced EF143813.1.
FEATURES             Location/Qualifiers
     source          1..8577
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    20..605
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             606..1322
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     LTR             1491..2082
                     /label=LTR
                     /note="long terminal repeat from Moloney murine leukemia
                     virus"
     CDS             2167..3024
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3198..3786
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    4074..4095
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        4110..4140
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4148..4164
                     /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)."
     primer_bind     4172..4188
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        4270..4573
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4578..4781
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     repeat_region   4782..4967
                     /label=5' LTR
                     /note="5' LTR"
     misc_feature    5020..5377
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus
                     (MMLV)"
     CDS             5442..5858
                     /codon_start=1
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag
                     gene lacking the start codon"
                     /translation="GQTVTTPLSLTLGHWKDVERIAHNQSVDVKKRRWVTFCSAEWPTF
                     NVGWPRDGTFNRDLITQVKIKVFSPGPHGHPDQVPYIVTWEALAFDPPPWVKPFVHPKP
                     PPPLPPSAPSLPLEPPRSTPPRSSLYPALTPSLGA"
     promoter        6205..6223
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             6653..6673
                     /codon_start=1
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
                     /translation="ENLYFQG"
     CDS             6680..6793
                     /codon_start=1
                     /label=SBP
                     /note="streptavidin-binding peptide"
                     /translation="MDEKTTGWRGGHVVEGLAGELEQLRARLEHHPQGQREP"
     protein_bind    6869..6993
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        7018..7048
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             7102..7758
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             8103..8405
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(8449..8573)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 ggccgctacg taaattccgc ccctctccct cccccccccc taacgttact ggccgaagcc
       61 gcttggaata aggccggtgt gcgtttgtct atatgttatt ttccaccata ttgccgtctt
      121 ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt gacgagcatt cctaggggtc
      181 tttcccctct cgccaaagga atgcaaggtc tgttgaatgt cgtgaaggaa gcagttcctc
      241 tggaagcttc ttgaagacaa acaacgtctg tagcgaccct ttgcaggcag cggaaccccc
      301 cacctggcga caggtgcctc tgcggccaaa agccacgtgt ataagataca cctgcaaagg
      361 cggcacaacc ccagtgccac gttgtgagtt ggatagttgt ggaaagagtc aaatggctct
      421 cctcaagcgt attcaacaag gggctgaagg atgcccagaa ggtaccccat tgtatgggat
      481 ctgatctggg gcctcggtgc acatgcttta catgtgttta gtcgaggtta aaaaacgtct
      541 aggccccccg aaccacgggg acgtggtttt cctttgaaaa acacgatgat aatatggcca
      601 caaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
      661 tggacggcga cgtgaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
      721 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
      781 ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca
      841 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
      901 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
      961 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     1021 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga
     1081 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc
     1141 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     1201 accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca
     1261 tggtcctgct ggagttcgtg accgccgccg ggatcactca cggcatggac gagctgtaca
     1321 agtaaagcgc cagcacagtg gtcgagatcc ggattagtcc aatttgttaa agacaggata
     1381 tcagtggtcc aggctctagt tttgactcaa caatatcacc agctgaagcc tatagagtac
     1441 gagccataga taaaataaaa gattttattt agtctccaga aaaagggggg aatgaaagac
     1501 cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc atggaaaaat
     1561 acataactga gaatagagaa gttcagatca aggtcaggaa cagatggaac agctgaatat
     1621 gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc aagaacagat
     1681 ggaacagctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca
     1741 gggccaagaa cagatggtcc ccagatgcgg tccagccctc agcagtttct agagaaccat
     1801 cagatgtttc cagggtgccc caaggacctg aaatgaccct gtgccttatt tgaactaacc
     1861 aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc cccgagctca ataaaagagc
     1921 ccacaacccc tcactcgggg cgccagtcct ccgattgact gagtcgcccg ggtacccgtg
     1981 tatccaataa accctcttgc agttgcatcc gacttgtggt ctcgctgttc cttgggaggg
     2041 tctcctctga gtgattgact acccgtcagc gggggtcttt cacacatgca gcatgtatca
     2101 aaattaattt ggtttttttt cttaagtatt tacattaaat ggccatagta ttgaaaaagg
     2161 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     2221 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     2281 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     2341 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     2401 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     2461 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     2521 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     2581 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     2641 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     2701 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     2761 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     2821 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     2881 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     2941 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     3001 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     3061 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     3121 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     3181 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     3241 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     3301 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg
     3361 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     3421 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     3481 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     3541 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     3601 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     3661 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     3721 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     3781 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     3841 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     3901 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     3961 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     4021 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg
     4081 agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg
     4141 tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc
     4201 aagcttgact agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
     4261 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
     4321 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
     4381 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
     4441 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
     4501 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
     4561 cgctattacc atgcatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
     4621 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
     4681 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
     4741 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tcaataaaag agcccacaac
     4801 ccctcactcg gcgcgccagt cctccgattg actgagtcgc ccgggtaccc gtgtatccaa
     4861 taaaccctct tgcagttgca tccgacttgt ggtctcgctg ttccttggga gggtctcctc
     4921 tgagtgattg actacccgtc agcgggggtc tttcatttgg gggctcgtcc gggatcggga
     4981 gacccctgcc cagggaccac cgacccacca ccgggaggta agctggccag caacttatct
     5041 gtgtctgtcc gattgtctag tgtctatgac tgattttatg cgcctgcgtc ggtactagtt
     5101 agctaactag ctctgtatct ggcggacccg tggtggaact gacgagttcg gaacacccgg
     5161 ccgcaaccct gggagacgtc ccagggactt cgggggccgt ttttgtggcc cgacctgagt
     5221 ccaaaaatcc cgatcgtttt ggactctttg gtgcaccccc cttagaggag ggatatgtgg
     5281 ttctggtagg agacgagaac ctaaaacagt tcccgcctcc gtctgaattt ttgctttcgg
     5341 tttgggaccg aagccgcgcc gcgcgtcttg tctgctgcag catcgttctg tgttgtctct
     5401 gtctgactgt gtttctgtat ttgtctgaaa atatgggccc gggccagact gttaccactc
     5461 ccttaagttt gaccttaggt cactggaaag atgtcgagcg gatcgctcac aaccagtcgg
     5521 tagatgtcaa gaagagacgt tgggttacct tctgctctgc agaatggcca acctttaacg
     5581 tcggatggcc gcgagacggc acctttaacc gagacctcat cacccaggtt aagatcaagg
     5641 tcttttcacc tggcccgcat ggacacccag accaggtccc ctacatcgtg acctgggaag
     5701 ccttggcttt tgacccccct ccctgggtca agccctttgt acaccctaag cctccgcctc
     5761 ctcttcctcc atccgccccg tctctccccc ttgaacctcc tcgttcgacc ccgcctcgat
     5821 cctcccttta tccagccctc actccttctc taggcgcccc catatggcca tatgagatct
     5881 tatatggggc acccccgccc cttgtaaact tccctgaccc tgacatgaca agagttacta
     5941 acagcccctc tctccaagct cacttacagg ctctctactt agtccagcac gaagtctgga
     6001 gacctctggc ggcagcctac caagaacaac tggaccgacc ggtggtacct cacccttacc
     6061 gagtcggcga cacagtgtgg gtccgccgac accagactaa gaacctagaa cctcgctgga
     6121 aaggacctta cacagtcctg ctgaccaccc ccaccgccct caaagtagac ggcatcgcag
     6181 cttggataca cgccgcccac gtgttaatac gactcactat agggaaggct gccgaccccg
     6241 ggggtggacc atcctctaga ctgccatgga attcatgggc acccccgcag tcaccaccta
     6301 caagctggtc attaacggca agactctgaa gggcgagacc accaccaagg ccgtggacgc
     6361 agaaaccgcg gaaaaggcgt tcaagcagta cgcgaacgac aacggggtgg acggagtctg
     6421 gacctacgat gacgctacaa aaacgttcac cgtgaccgag gttaacaccc ccgctgtgac
     6481 tacgtacaag ctggtgatca atgggaagac cctgaagggc gagaccacca cgaaagctgt
     6541 agacgccgag acagccgaga aggccttcaa gcagtacgcc aatgacaacg gcgtggatgg
     6601 cgtgtggacc tatgacgacg ccacgaagac attcactgtg actgagatcg atgagaacct
     6661 ctacttccag ggcggcagta tggacgagaa gaccaccggc tggagaggcg gccacgtggt
     6721 cgagggactg gccggtgagc tggagcagct acgggcccgc cttgagcacc acccccaggg
     6781 gcagcgcgag ccgggccggc cggcgcgcca tatggctagc ggccatcaaa caagtttgta
     6841 ctcgagggcc ggcgcgccgc ggccatcaac aagtttgtac aaaaaagctg aacgagaaac
     6901 gtaaaatgat ataaatatca atatattaaa ttagattttg cataaaaaac agactacata
     6961 atactgtaaa acacaacata tccagtcact atggcggccg cattaggcac cccaggcttt
     7021 acactttatg cttccggctc gtataatgtg tggattttga gttaggatcc gtcgagattt
     7081 tcaggagcta aggaagctaa aatggagaaa aaaatcactg gatataccac cgttgatata
     7141 tcccaatggc atcgtaaaga acattttgag gcatttcagt cagttgctca atgtacctat
     7201 aaccagaccg ttcagctgga tattacggcc tttttaaaga ccgtaaagaa aaataagcac
     7261 aagttttatc cggcctttat tcacattctt gcccgcctga tgaatgctca tccggaattc
     7321 cgtatggcaa tgaaagacgg tgagctggtg atatgggata gtgttcaccc ttgttacacc
     7381 gttttccatg agcaaactga aacgttttca tcgctctgga gtgaatacca cgacgatttc
     7441 cggcagtttc tacacatata ttcgcaagat gtggcgtgtt acggtgaaaa cctggcctat
     7501 ttccctaaag ggtttattga gaatatgttt ttcgtctcag ccaatccctg ggtgagtttc
     7561 accagttttg atttaaacgt ggccaatatg gacaacttct tcgcccccgt tttcaccatg
     7621 ggcaaatatt atacgcaagg cgacaaggtg ctgatgccgc tggcgattca ggttcatcat
     7681 gccgtctgtg atggcttcca tgtcggcaga atgcttaatg aattacaaca gtactgcgat
     7741 gagtggcagg gcggggcgta atctagagga tccggcttac taaaagccag ataacagtat
     7801 gcgtatttgc gcgctgattt ttgcggtata agaatatata ctgatatgta tacccgaagt
     7861 atgtcaaaaa gaggtgtgct atgaagcagc gtattacagt gacagttgac agcgacagct
     7921 atcagttgct caaggcatat atgatgtcaa tatctccggt ctggtaagca caaccatgca
     7981 gaatgaagcc cgtcgtctgc gtgccgaacg ctggaaagcg gaaaatcagg aagggatggc
     8041 tgaggtcgcc cggtttattg aaatgaacgg ctcttttgct gacgagaaca gggactggtg
     8101 aaatgcagtt taaggtttac acctataaaa gagagagccg ttatcgtctg tttgtggatg
     8161 tacagagtga tattattgac acgcccgggc gacggatggt gatccccctg gccagtgcac
     8221 gtctgctgtc agataaagtc tcccgtgaac tttacccggt ggtgcatatc ggggatgaaa
     8281 gctggcgcat gatgaccacc gatatggcca gtgtgccggt ctccgttatc ggggaagaag
     8341 tggctgatct cagccaccgc gaaaatgaca tcaaaaacgc cattaacctg atgttctggg
     8401 gaatataaat gtcaggctcc cttatacaca gccagtctgc aggtcgacca tagtgactgg
     8461 atatgttgtg ttttacagta ttatgtagtc tgttttttat gcaaaatcta atttaatata
     8521 ttgatattta tatcatttta cgtttctcgt tcagctttct tgtacaaagt ggttgat
//
