LOCUS       sequence-169838-        6529 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 6529)
  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 6529)
  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-C5(GST)
FEATURES             Location/Qualifiers
     source          1..6529
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             540..845
                     /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     724..743
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(886..1010)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             1018..1671
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     1629..1651
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             1678..1701
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     polyA_signal    1782..1916
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1806..1825)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1860..1879
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2029..2049)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2029..2047)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2068..2084)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2068..2084)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2081..2103)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2092..2108
                     /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(2116..2122,2123..2140,2141..2146))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2161..2182
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2299..2316)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2470..3058)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2550..2569)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3229..4020,4021..4089))
                     /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     3852..3871
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4090..4194)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4220..4675)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4357..4378)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4569..4588
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4746..5730
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        5765..5783
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     5765..5782
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    5823..5947
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(5984..5989,5990..6007,6008..6014)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             join(6068..6529,1..198)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(6134..6153)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 agttttgatt taaacgtggc caatatggac aacttcttcg cccccgtttt caccatgggc
       61 aaatattata cgcaaggcga caaggtgctg atgccgctgg cgattcaggt tcatcatgcc
      121 gtttgtgatg gcttccatgt cggcagaatg cttaatgaat tacaacagta ctgcgatgag
      181 tggcagggcg gggcgtaaac gcgtggatcc ggcttactaa aagccagata acagtatgcg
      241 tatttgcgcg ctgatttttg cggtataaga atatatactg atatgtatac ccgaagtatg
      301 tcaaaaagag gtatgctatg aagcagcgta ttacagtgac agttgacagc gacagctatc
      361 agttgctcaa ggcatatatg atgtcaatat ctccggtctg gtaagcacaa ccatgcagaa
      421 tgaagcccgt cgtctgcgtg ccgaacgctg gaaagcggaa aatcaggaag ggatggctga
      481 ggtcgcccgg tttattgaaa tgaacggctc ttttgctgac gagaacaggg gctggtgaaa
      541 tgcagtttaa ggtttacacc tataaaagag agagccgtta tcgtctgttt gtggatgtac
      601 agagtgatat tattgacacg cccgggcgac ggatggtgat ccccctggcc agtgcacgtc
      661 tgctgtcaga taaagtctcc cgtgaacttt acccggtggt gcatatcggg gatgaaagct
      721 ggcgcatgat gaccaccgat atggccagtg tgccggtctc cgttatcggg gaagaagtgg
      781 ctgatctcag ccaccgcgaa aatgacatca aaaacgccat taacctgatg ttctggggaa
      841 tataaatgtc aggctccctt atacacagcc agtctgcagg tcgaccatag tgactggata
      901 tgttgtgttt tacagtatta tgtagtctgt tttttatgca aaatctaatt taatatattg
      961 atatttatat cattttacgt ttctcgttca gctttcttgt acaaagtggt cgagctcatg
     1021 tcccctatac taggttattg gaaaattaag ggccttgtgc aacccactcg acttcttttg
     1081 gaatatcttg aagaaaaata tgaagagcat ttgtatgagc gcgatgaagg tgataaatgg
     1141 cgaaacaaaa agtttgaatt gggtttggag tttcccaatc ttccttatta tattgatggt
     1201 gatgttaaat taacacagtc tatggccatc atacgttata tagctgacaa gcacaacatg
     1261 ttgggtggtt gtccaaaaga gcgtgcagag atttcaatgc ttgaaggagc ggttttggat
     1321 attagatacg gtgtttcgag aattgcatat agtaaagact ttgaaactct caaagttgat
     1381 tttcttagca agctacctga aatgctgaaa atgttcgaag atcgtttatg tcataaaaca
     1441 tatttaaatg gtgatcatgt aacccatcct gacttcatgt tgtatgacgc tcttgatgtt
     1501 gttttataca tggacccaat gtgcctggat gcgttcccaa aattagtttg ttttaaaaaa
     1561 cgtattgaag ctatcccaca aattgataag tacttgaaat ccagcaagta tatagcatgg
     1621 cctttgcagg gctggcaagc cacgtttggt ggtggcgacc atcctccaaa atcggatctg
     1681 gaagttctgt tccaggggcc cctgggatcc ccggaattcc cgggtcgact cgagcggccg
     1741 catcgtgact gatctagaac tatagtgagt cgtattacgt agatccagac atgataagat
     1801 acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg
     1861 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca
     1921 acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaat
     1981 tcgcggccgc ggcgccaatg cattgggccc ggtacccagc ttttgttccc tttagtgagg
     2041 gttaattgcg cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     2101 gctcacaatt ccacacaaca tacgagccgg gagcataaag tgtaaagcct ggggtgccta
     2161 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     2221 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     2281 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     2341 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     2401 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     2461 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     2521 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     2581 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     2641 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     2701 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     2761 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     2821 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     2881 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     2941 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     3001 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     3061 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     3121 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     3181 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     3241 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     3301 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     3361 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     3421 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     3481 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     3541 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     3601 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     3661 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     3721 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     3781 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     3841 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     3901 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     3961 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     4021 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     4081 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     4141 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     4201 tccccgaaaa gtgccaccta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     4261 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     4321 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     4381 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     4441 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     4501 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     4561 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     4621 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat
     4681 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
     4741 cgccagtcga ccgccaattc aatatggcgt atatggactc atgccaattc aatatggtgg
     4801 atctggacct gtgccaattc aatatggcgt atatggactc gtgccaattc aatatggtgg
     4861 atctggaccc cagccaattc aatatggcgg acttggcacc atgccaattc aatatggcgg
     4921 acctggcact gtgccaactg gggaggggtc tacttggcac ggtgccaagt ttgaggaggg
     4981 gtcttggccc tgtgccaagt ccgccatatt gaattggcat ggtgccaata atggcggcca
     5041 tattggctat atgccaggat caatatatag gcaatatcca atatggccct atgccaatat
     5101 ggctattggc caggttcaat actatgtatt ggccctatgc catatagtat tccatatatg
     5161 ggttttccta ttgacgtaga tagcccctcc caatgggcgg tcccatatac catatatggg
     5221 gcttcctaat accgcccata gccactcccc cattgacgtc aatggtctct atatatggtc
     5281 tttcctattg acgtcatatg ggcggtccta ttgacgtata tggcgcctcc cccattgacg
     5341 tcaattacgg taaatggccc gcctggctca atgcccattg acgtcaatag gaccacccac
     5401 cattgacgtc aatgggatgg ctcattgccc attcatatcc gttctcacgc cccctattga
     5461 cgtcaatgac ggtaaatggc ccacttggca gtacatcaat atctattaat agtaacttgg
     5521 caagtacatt actattggaa gtacgccagg gtacattggc agtactccca ttgacgtcaa
     5581 tggcggtaaa tggcccgcga tggctgccaa gtacatcccc attgacgtca atggggaggg
     5641 gcaatgacgc aaatgggcgt tccattgacg taaatgggcg gtaggcgtgc ctaatgggag
     5701 gtctatataa gcaatgctcg tttagggaac cgccattctg cctggggacg tcggagcaag
     5761 cttgatttag gtgacactat agaatacaag ctacttgttc tttttgcagg atccggccgg
     5821 ccacaagttt gtacaaaaaa gctgaacgag aaacgtaaaa tgatataaat atcaatatat
     5881 taaattagat tttgcataaa aaacagacta cataatactg taaaacacaa catatccagt
     5941 cactatggcg gcccctcgag gggccgcatt aggcacccca ggctttacac tttatgcttc
     6001 cggctcgtat aatgtgtgga ttttgagtta ggatccgtcg agattttcag gagctaagga
     6061 agctaaaatg gagaaaaaaa tcactggata taccaccgtt gatatatccc aatggcatcg
     6121 taaagaacat tttgaggcat ttcagtcagt tgctcaatgt acctataacc agaccgttca
     6181 gctggatatt acggcctttt taaagaccgt aaagaaaaat aagcacaagt tttatccggc
     6241 ctttattcac attcttgccc gcctgatgaa tgctcatccg gaattccgta tggcaatgaa
     6301 agacggtgag ctggtgatat gggatagtgt tcacccttgt tacaccgttt tccatgagca
     6361 aactgaaacg ttttcatcgc tctggagtga ataccacgac gatttccggc agtttctaca
     6421 catatattcg caagatgtgg cgtgttacgg tgaaaacctg gcctatttcc ctaaagggtt
     6481 tattgagaat atgtttttcg tctcagccaa tccctgggtg agtttcacc
//
