LOCUS       sequence-169829-        5787 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 5787)
  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 5787)
  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: pGCS1
FEATURES             Location/Qualifiers
     source          1..5787
                     /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"
     protein_bind    93..217
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(254..259,260..277,278..284)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             338..997
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(404..423)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             1339..1644
                     /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     1523..1542
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(1685..1809)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     polyA_signal    1839..1973
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1863..1882)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1917..1936
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2086..2106)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2086..2104)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2125..2141)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2125..2141)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2138..2160)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2149..2165
                     /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(2173..2179,2180..2197,2198..2203))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2218..2239
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2356..2373)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2527..3115)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2607..2626)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3286..4077,4078..4146))
                     /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     3909..3928
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4147..4251)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4277..4732)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4414..4435)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4626..4645
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4803..5787
                     /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 atccggccgg ccacaagttt gtacaaaaaa gctgaacgag
      121 aaacgtaaaa tgatataaat atcaatatat taaattagat tttgcataaa aaacagacta
      181 cataatactg taaaacacaa catatccagt cactatggcg gcccctcgag gggccgcatt
      241 aggcacccca ggctttacac tttatgcttc cggctcgtat aatgtgtgga ttttgagtta
      301 ggatccgtcg agattttcag gagctaagga agctaaaatg gagaaaaaaa tcactggata
      361 taccaccgtt gatatatccc aatggcatcg taaagaacat tttgaggcat ttcagtcagt
      421 tgctcaatgt acctataacc agaccgttca gctggatatt acggcctttt taaagaccgt
      481 aaagaaaaat aagcacaagt tttatccggc ctttattcac attcttgccc gcctgatgaa
      541 tgctcatccg gaattccgta tggcaatgaa agacggtgag ctggtgatat gggatagtgt
      601 tcacccttgt tacaccgttt tccatgagca aactgaaacg ttttcatcgc tctggagtga
      661 ataccacgac gatttccggc agtttctaca catatattcg caagatgtgg cgtgttacgg
      721 tgaaaacctg gcctatttcc ctaaagggtt tattgagaat atgtttttcg tctcagccaa
      781 tccctgggtg agtttcacca gttttgattt aaacgtggcc aatatggaca acttcttcgc
      841 ccccgttttc accatgggca aatattatac gcaaggcgac aaggtgctga tgccgctggc
      901 gattcaggtt catcatgccg tttgtgatgg cttccatgtc ggcagaatgc ttaatgaatt
      961 acaacagtac tgcgatgagt ggcagggcgg ggcgtaaacg cgtggatccg gcttactaaa
     1021 agccagataa cagtatgcgt atttgcgcgc tgatttttgc ggtataagaa tatatactga
     1081 tatgtatacc cgaagtatgt caaaaagagg tatgctatga agcagcgtat tacagtgaca
     1141 gttgacagcg acagctatca gttgctcaag gcatatatga tgtcaatatc tccggtctgg
     1201 taagcacaac catgcagaat gaagcccgtc gtctgcgtgc cgaacgctgg aaagcggaaa
     1261 atcaggaagg gatggctgag gtcgcccggt ttattgaaat gaacggctct tttgctgacg
     1321 agaacagggg ctggtgaaat gcagtttaag gtttacacct ataaaagaga gagccgttat
     1381 cgtctgtttg tggatgtaca gagtgatatt attgacacgc ccgggcgacg gatggtgatc
     1441 cccctggcca gtgcacgtct gctgtcagat aaagtctccc gtgaacttta cccggtggtg
     1501 catatcgggg atgaaagctg gcgcatgatg accaccgata tggccagtgt gccggtctcc
     1561 gttatcgggg aagaagtggc tgatctcagc caccgcgaaa atgacatcaa aaacgccatt
     1621 aacctgatgt tctggggaat ataaatgtca ggctccctta tacacagcca gtctgcaggt
     1681 cgaccatagt gactggatat gttgtgtttt acagtattat gtagtctgtt ttttatgcaa
     1741 aatctaattt aatatattga tatttatatc attttacgtt tctcgttcag ctttcttgta
     1801 caaagtggtt ctagaactat agtgagtcgt attacgtaga tccagacatg ataagataca
     1861 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
     1921 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
     1981 acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaattcg
     2041 cggccgcggc gccaatgcat tgggcccggt acccagcttt tgttcccttt agtgagggtt
     2101 aattgcgcgc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     2161 cacaattcca cacaacatac gagccgggag cataaagtgt aaagcctggg gtgcctaatg
     2221 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     2281 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     2341 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     2401 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     2461 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     2521 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     2581 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     2641 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     2701 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     2761 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     2821 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     2881 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     2941 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     3001 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     3061 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     3121 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     3181 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     3241 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     3301 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     3361 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     3421 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     3481 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     3541 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     3601 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     3661 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     3721 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     3781 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     3841 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     3901 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     3961 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     4021 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     4081 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     4141 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     4201 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     4261 ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
     4321 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     4381 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     4441 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     4501 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     4561 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     4621 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     4681 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg
     4741 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
     4801 cagtcgaccg ccaattcaat atggcgtata tggactcatg ccaattcaat atggtggatc
     4861 tggacctgtg ccaattcaat atggcgtata tggactcgtg ccaattcaat atggtggatc
     4921 tggaccccag ccaattcaat atggcggact tggcaccatg ccaattcaat atggcggacc
     4981 tggcactgtg ccaactgggg aggggtctac ttggcacggt gccaagtttg aggaggggtc
     5041 ttggccctgt gccaagtccg ccatattgaa ttggcatggt gccaataatg gcggccatat
     5101 tggctatatg ccaggatcaa tatataggca atatccaata tggccctatg ccaatatggc
     5161 tattggccag gttcaatact atgtattggc cctatgccat atagtattcc atatatgggt
     5221 tttcctattg acgtagatag cccctcccaa tgggcggtcc catataccat atatggggct
     5281 tcctaatacc gcccatagcc actcccccat tgacgtcaat ggtctctata tatggtcttt
     5341 cctattgacg tcatatgggc ggtcctattg acgtatatgg cgcctccccc attgacgtca
     5401 attacggtaa atggcccgcc tggctcaatg cccattgacg tcaataggac cacccaccat
     5461 tgacgtcaat gggatggctc attgcccatt catatccgtt ctcacgcccc ctattgacgt
     5521 caatgacggt aaatggccca cttggcagta catcaatatc tattaatagt aacttggcaa
     5581 gtacattact attggaagta cgccagggta cattggcagt actcccattg acgtcaatgg
     5641 cggtaaatgg cccgcgatgg ctgccaagta catccccatt gacgtcaatg gggaggggca
     5701 atgacgcaaa tgggcgttcc attgacgtaa atgggcggta ggcgtgccta atgggaggtc
     5761 tatataagca atgctcgttt agggaac
//
