LOCUS       40924_14355        8123 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Gateway(R) destination vector for high-level expression of 
            N-terminally GST-tagged proteins in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8123)
  AUTHORS   Invitrogen (Life Technologies)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 8123)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8123
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        15..318
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        319..522
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             632..1285
                     /codon_start=1
                     /label=GST
                     /note="glutathione S-transferase from Schistosoma
                     japonicum"
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     protein_bind    1313..1437
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        1462..1492
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             1546..2202
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             2547..2849
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(2893..3017)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(3050..3068)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3079..3095)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    3500..3634
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      3760..4215
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4334..4691
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             4742..5533
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    5600..5648
                     /label=poly(A) signal
                     /note="synthetic polyadenylation signal"
     promoter        5954..6058
                     /label=AmpR promoter
     CDS             6059..6916
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      7090..7678
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cattagatgc atgtcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
       61 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
      121 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
      181 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
      241 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
      301 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg
      361 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
      421 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
      481 ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctcgtttagt gaaccgtcag
      541 atcgcctgga gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc
      601 agcctccgga ctctagccta ggccgcggac catgtcccct atactaggtt attggaaaat
      661 taagggcctt gtgcaaccca ctcgacttct tttggaatat cttgaagaaa aatatgaaga
      721 gcatttgtat gagcgcgatg aaggtgataa atggcgaaac aaaaagtttg aattgggttt
      781 ggagtttccc aatcttcctt attatattga tggtgatgtt aaattaacac agtctatggc
      841 catcatacgt tatatagctg acaagcacaa catgttgggt ggttgtccaa aagagcgtgc
      901 agagatttca atgcttgaag gagcggtttt ggatattaga tacggtgttt cgagaattgc
      961 atatagtaaa gactttgaaa ctctcaaagt tgattttctt agcaagctac ctgaaatgct
     1021 gaaaatgttc gaagatcgtt tatgtcataa aacatattta aatggtgatc atgtaaccca
     1081 tcctgacttc atgttgtatg acgctcttga tgttgtttta tacatggacc caatgtgcct
     1141 ggatgcgttc ccaaaattag tttgttttaa aaaacgtatt gaagctatcc cacaaattga
     1201 taagtacttg aaatccagca agtatatagc atggcctttg cagggctggc aagccacgtt
     1261 tggtggtggc gaccatcctc caaaatcgga tctggttccg cgttctagat caacaagttt
     1321 gtacaaaaaa gctgaacgag aaacgtaaaa tgatataaat atcaatatat taaattagat
     1381 tttgcataaa aaacagacta cataatactg taaaacacaa catatccagt cactatggcg
     1441 gccgcattag gcaccccagg ctttacactt tatgcttccg gctcgtataa tgtgtggatt
     1501 ttgagttagg atccggcgag attttcagga gctaaggaag ctaaaatgga gaaaaaaatc
     1561 actggatata ccaccgttga tatatcccaa tggcatcgta aagaacattt tgaggcattt
     1621 cagtcagttg ctcaatgtac ctataaccag accgttcagc tggatattac ggccttttta
     1681 aagaccgtaa agaaaaataa gcacaagttt tatccggcct ttattcacat tcttgcccgc
     1741 ctgatgaatg ctcatccgga attccgtatg gcaatgaaag acggtgagct ggtgatatgg
     1801 gatagtgttc acccttgtta caccgttttc catgagcaaa ctgaaacgtt ttcatcgctc
     1861 tggagtgaat accacgacga tttccggcag tttctacaca tatattcgca agatgtggcg
     1921 tgttacggtg aaaacctggc ctatttccct aaagggttta ttgagaatat gtttttcgtc
     1981 tcagccaatc cctgggtgag tttcaccagt tttgatttaa acgtggccaa tatggacaac
     2041 ttcttcgccc ccgttttcac catgggcaaa tattatacgc aaggcgacaa ggtgctgatg
     2101 ccgctggcga ttcaggttca tcatgccgtc tgtgatggct tccatgtcgg cagaatgctt
     2161 aatgaattac aacagtactg cgatgagtgg cagggcgggg cgtaaagatc tggatccggc
     2221 ttactaaaag ccagataaca gtatgcgtat ttgcgcgctg atttttgcgg tataagaata
     2281 tatactgata tgtatacccg aagtatgtca aaaagaggtg tgctatgaag cagcgtatta
     2341 cagtgacagt tgacagcgac agctatcagt tgctcaaggc atatatgatg tcaatatctc
     2401 cggtctggta agcacaacca tgcagaatga agcccgtcgt ctgcgtgccg aacgctggaa
     2461 agcggaaaat caggaaggga tggctgaggt cgcccggttt attgaaatga acggctcttt
     2521 tgctgacgag aacagggact ggtgaaatgc agtttaaggt ttacacctat aaaagagaga
     2581 gccgttatcg tctgtttgtg gatgtacaga gtgatattat tgacacgccc gggcgacgga
     2641 tggtgatccc cctggccagt gcacgtctgc tgtcagataa agtctcccgt gaactttacc
     2701 cggtggtgca tatcggggat gaaagctggc gcatgatgac caccgatatg gccagtgtgc
     2761 cggtctccgt tatcggggaa gaagtggctg atctcagcca ccgcgaaaat gacatcaaaa
     2821 acgccattaa cctgatgttc tggggaatat aaatgtcagg ctcccttata cacagccagt
     2881 ctgcaggtcg accatagtga ctggatatgt tgtgttttac agtattatgt agtctgtttt
     2941 ttatgcaaaa tctaatttaa tatattgata tttatatcat tttacgtttc tcgttcagct
     3001 ttcttgtaca aagtggttga tcgcgtgcat gcgacgtcat agctctctcc ctatagtgag
     3061 tcgtattata agctaggcac tggccgtcgt tttacaacgt cgtgactggg aaaactgcta
     3121 gcttgggatc tttgtgaagg aaccttactt ctgtggtgtg acataattgg acaaactacc
     3181 tacagagatt taaagctcta aggtaaatat aaaattttta agtgtataat gtgttaaact
     3241 agctgcatat gcttgctgct tgagagtttt gcttactgag tatgatttat gaaaatatta
     3301 tacacaggag ctagtgattc taattgtttg tgtattttag attcacagtc ccaaggctca
     3361 tttcaggccc ctcagtcctc acagtctgtt catgatcata atcagccata ccacatttgt
     3421 agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat
     3481 gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa
     3541 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     3601 caaactcatc aatgtatctt atcatgtctg gatcgatcct gcattaatga atcggccaac
     3661 gcgcggggag aggcggtttg cgtattggct ggcgtaatag cgaagaggcc cgcaccgatc
     3721 gcccttccca acagttgcgc agcctgaatg gcgaatggga cgcgccctgt agcggcgcat
     3781 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     3841 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     3901 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     3961 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     4021 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     4081 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     4141 cctattggtt aaaaaatgag ctgatttaac aaatatttaa cgcgaatttt aacaaaatat
     4201 taacgtttac aatttcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     4261 caccgcatac gcggatctgc gcagcaccat ggcctgaaat aacctctgaa agaggaactt
     4321 ggttaggtac cttctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg
     4381 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     4441 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     4501 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     4561 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     4621 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     4681 gcttttgcaa aaagcttgat tcttctgaca caacagtctc gaacttaagg ctagagccac
     4741 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     4801 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     4861 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     4921 gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     4981 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     5041 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     5101 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     5161 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     5221 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga
     5281 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     5341 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     5401 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     5461 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     5521 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     5581 ctgccatcac gatggccgca ataaaatatc tttattttca ttacatctgt gtgttggttt
     5641 tttgtgtgaa tcgatagcga taaggatccg cgtatggtgc actctcagta caatctgctc
     5701 tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg
     5761 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     5821 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg
     5881 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     5941 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     6001 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     6061 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     6121 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     6181 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     6241 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     6301 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     6361 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     6421 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     6481 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     6541 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     6601 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     6661 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     6721 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     6781 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     6841 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     6901 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     6961 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     7021 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     7081 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     7141 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     7201 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
     7261 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     7321 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     7381 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     7441 gcgaacgacc tacaccgaac tgagatacct acagcgtgag cattgagaaa gcgccacgct
     7501 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     7561 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     7621 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     7681 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     7741 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     7801 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     7861 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagagcttg
     7921 caattcgcgc gtttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     7981 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     8041 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     8101 gcgtagtacg aggccctttc act
//
