LOCUS       sequence-169839-        6457 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway destination vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6457)
  AUTHORS   Wang HY, Li Y, Xue T, Cheng N, Du HN
  TITLE     Construction of a series of pCS2+ backbone-based Gateway vectors for
            overexpressing various tagged proteins in vertebrates.
  JOURNAL   Acta Biochim Biophys Sin (Shanghai). 2016 Oct 26.
  PUBMED    27797719
REFERENCE   2  (bases 1 to 6457)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Acta 
            Biochim Biophys Sin (Shanghai). 2016 Oct 26."
COMMENT     Sequence Label: pGCS-N5(GST)
FEATURES             Location/Qualifiers
     source          1..6457
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             91..744
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     702..724
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     protein_bind    763..887
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(924..929,930..947,948..954)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             1008..1667
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1074..1093)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             2009..2314
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     2193..2212
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(2355..2479)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     polyA_signal    2509..2643
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2533..2552)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2587..2606
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2756..2776)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2756..2774)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2795..2811)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2795..2811)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2808..2830)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2819..2835
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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(join(2843..2849,2850..2867,2868..2873))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2888..2909
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3026..3043)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3197..3785)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3277..3296)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3956..4747,4748..4816))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4579..4598
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4817..4921)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4947..5402)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5084..5105)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5296..5315
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5473..6457
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atccgccacc atgtccccta tactaggtta ttggaaaatt
      121 aagggccttg tgcaacccac tcgacttctt ttggaatatc ttgaagaaaa atatgaagag
      181 catttgtatg agcgcgatga aggtgataaa tggcgaaaca aaaagtttga attgggtttg
      241 gagtttccca atcttcctta ttatattgat ggtgatgtta aattaacaca gtctatggcc
      301 atcatacgtt atatagctga caagcacaac atgttgggtg gttgtccaaa agagcgtgca
      361 gagatttcaa tgcttgaagg agcggttttg gatattagat acggtgtttc gagaattgca
      421 tatagtaaag actttgaaac tctcaaagtt gattttctta gcaagctacc tgaaatgctg
      481 aaaatgttcg aagatcgttt atgtcataaa acatatttaa atggtgatca tgtaacccat
      541 cctgacttca tgttgtatga cgctcttgat gttgttttat acatggaccc aatgtgcctg
      601 gatgcgttcc caaaattagt ttgttttaaa aaacgtattg aagctatccc acaaattgat
      661 aagtacttga aatccagcaa gtatatagca tggcctttgc agggctggca agccacgttt
      721 ggtggtggcg accatcctcc aaaatcggat ctgggccggc caacaagttt gtacaaaaaa
      781 gctgaacgag aaacgtaaaa tgatataaat atcaatatat taaattagat tttgcataaa
      841 aaacagacta cataatactg taaaacacaa catatccagt cactatggcg gcccctcgag
      901 gggccgcatt aggcacccca ggctttacac tttatgcttc cggctcgtat aatgtgtgga
      961 ttttgagtta ggatccgtcg agattttcag gagctaagga agctaaaatg gagaaaaaaa
     1021 tcactggata taccaccgtt gatatatccc aatggcatcg taaagaacat tttgaggcat
     1081 ttcagtcagt tgctcaatgt acctataacc agaccgttca gctggatatt acggcctttt
     1141 taaagaccgt aaagaaaaat aagcacaagt tttatccggc ctttattcac attcttgccc
     1201 gcctgatgaa tgctcatccg gaattccgta tggcaatgaa agacggtgag ctggtgatat
     1261 gggatagtgt tcacccttgt tacaccgttt tccatgagca aactgaaacg ttttcatcgc
     1321 tctggagtga ataccacgac gatttccggc agtttctaca catatattcg caagatgtgg
     1381 cgtgttacgg tgaaaacctg gcctatttcc ctaaagggtt tattgagaat atgtttttcg
     1441 tctcagccaa tccctgggtg agtttcacca gttttgattt aaacgtggcc aatatggaca
     1501 acttcttcgc ccccgttttc accatgggca aatattatac gcaaggcgac aaggtgctga
     1561 tgccgctggc gattcaggtt catcatgccg tttgtgatgg cttccatgtc ggcagaatgc
     1621 ttaatgaatt acaacagtac tgcgatgagt ggcagggcgg ggcgtaaacg cgtggatccg
     1681 gcttactaaa agccagataa cagtatgcgt atttgcgcgc tgatttttgc ggtataagaa
     1741 tatatactga tatgtatacc cgaagtatgt caaaaagagg tatgctatga agcagcgtat
     1801 tacagtgaca gttgacagcg acagctatca gttgctcaag gcatatatga tgtcaatatc
     1861 tccggtctgg taagcacaac catgcagaat gaagcccgtc gtctgcgtgc cgaacgctgg
     1921 aaagcggaaa atcaggaagg gatggctgag gtcgcccggt ttattgaaat gaacggctct
     1981 tttgctgacg agaacagggg ctggtgaaat gcagtttaag gtttacacct ataaaagaga
     2041 gagccgttat cgtctgtttg tggatgtaca gagtgatatt attgacacgc ccgggcgacg
     2101 gatggtgatc cccctggcca gtgcacgtct gctgtcagat aaagtctccc gtgaacttta
     2161 cccggtggtg catatcgggg atgaaagctg gcgcatgatg accaccgata tggccagtgt
     2221 gccggtctcc gttatcgggg aagaagtggc tgatctcagc caccgcgaaa atgacatcaa
     2281 aaacgccatt aacctgatgt tctggggaat ataaatgtca ggctccctta tacacagcca
     2341 gtctgcaggt cgaccatagt gactggatat gttgtgtttt acagtattat gtagtctgtt
     2401 ttttatgcaa aatctaattt aatatattga tatttatatc attttacgtt tctcgttcag
     2461 ctttcttgta caaagtggtt ctagaactat agtgagtcgt attacgtaga tccagacatg
     2521 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
     2581 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
     2641 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt
     2701 ttttaattcg cggccgcggc gccaatgcat tgggcccggt acccagcttt tgttcccttt
     2761 agtgagggtt aattgcgcgc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     2821 gttatccgct cacaattcca cacaacatac gagccgggag cataaagtgt aaagcctggg
     2881 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     2941 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     3001 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     3061 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     3121 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     3181 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     3241 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     3301 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     3361 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     3421 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     3481 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     3541 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     3601 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     3661 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     3721 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     3781 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     3841 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     3901 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     3961 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     4021 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     4081 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     4141 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     4201 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     4261 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     4321 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     4381 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     4441 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     4501 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     4561 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     4621 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     4681 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     4741 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     4801 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     4861 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     4921 gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt taaaattcgc
     4981 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     5041 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     5101 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     5161 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     5221 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     5281 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     5341 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     5401 gtcccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
     5461 gctattacgc cagtcgaccg ccaattcaat atggcgtata tggactcatg ccaattcaat
     5521 atggtggatc tggacctgtg ccaattcaat atggcgtata tggactcgtg ccaattcaat
     5581 atggtggatc tggaccccag ccaattcaat atggcggact tggcaccatg ccaattcaat
     5641 atggcggacc tggcactgtg ccaactgggg aggggtctac ttggcacggt gccaagtttg
     5701 aggaggggtc ttggccctgt gccaagtccg ccatattgaa ttggcatggt gccaataatg
     5761 gcggccatat tggctatatg ccaggatcaa tatataggca atatccaata tggccctatg
     5821 ccaatatggc tattggccag gttcaatact atgtattggc cctatgccat atagtattcc
     5881 atatatgggt tttcctattg acgtagatag cccctcccaa tgggcggtcc catataccat
     5941 atatggggct tcctaatacc gcccatagcc actcccccat tgacgtcaat ggtctctata
     6001 tatggtcttt cctattgacg tcatatgggc ggtcctattg acgtatatgg cgcctccccc
     6061 attgacgtca attacggtaa atggcccgcc tggctcaatg cccattgacg tcaataggac
     6121 cacccaccat tgacgtcaat gggatggctc attgcccatt catatccgtt ctcacgcccc
     6181 ctattgacgt caatgacggt aaatggccca cttggcagta catcaatatc tattaatagt
     6241 aacttggcaa gtacattact attggaagta cgccagggta cattggcagt actcccattg
     6301 acgtcaatgg cggtaaatgg cccgcgatgg ctgccaagta catccccatt gacgtcaatg
     6361 gggaggggca atgacgcaaa tgggcgttcc attgacgtaa atgggcggta ggcgtgccta
     6421 atgggaggtc tatataagca atgctcgttt agggaac
//
