LOCUS       sequence-169833-        5816 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 5816)
  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 5816)
  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: pGCS2
FEATURES             Location/Qualifiers
     source          1..5816
                     /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    1868..2002
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1892..1911)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1946..1965
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2115..2135)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2115..2133)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2154..2170)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2154..2170)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2167..2189)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2178..2194
                     /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(2202..2208,2209..2226,2227..2232))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2247..2268
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2385..2402)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2556..3144)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2636..2655)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3315..4106,4107..4175))
                     /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     3938..3957
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4176..4280)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4306..4761)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4443..4464)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4655..4674
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4832..5816
                     /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 caaagtggtg aagacatgat ccaagctagc cgtcttcttc tagaactata gtgagtcgta
     1861 ttacgtagat ccagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     1921 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     1981 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     2041 gggggaggtg tgggaggttt tttaattcgc ggccgcggcg ccaatgcatt gggcccggta
     2101 cccagctttt gttcccttta gtgagggtta attgcgcgct tggcgtaatc atggtcatag
     2161 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccgggagc
     2221 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     2281 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     2341 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     2401 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     2461 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     2521 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     2581 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     2641 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     2701 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     2761 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     2821 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     2881 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     2941 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     3001 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     3061 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     3121 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     3181 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     3241 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     3301 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     3361 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     3421 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     3481 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     3541 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     3601 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     3661 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     3721 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     3781 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     3841 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     3901 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     3961 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     4021 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     4081 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     4141 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     4201 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     4261 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta
     4321 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     4381 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     4441 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     4501 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     4561 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     4621 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     4681 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     4741 atgcgccgct acagggcgcg tcccattcgc cattcaggct gcgcaactgt tgggaagggc
     4801 gatcggtgcg ggcctcttcg ctattacgcc agtcgaccgc caattcaata tggcgtatat
     4861 ggactcatgc caattcaata tggtggatct ggacctgtgc caattcaata tggcgtatat
     4921 ggactcgtgc caattcaata tggtggatct ggaccccagc caattcaata tggcggactt
     4981 ggcaccatgc caattcaata tggcggacct ggcactgtgc caactgggga ggggtctact
     5041 tggcacggtg ccaagtttga ggaggggtct tggccctgtg ccaagtccgc catattgaat
     5101 tggcatggtg ccaataatgg cggccatatt ggctatatgc caggatcaat atataggcaa
     5161 tatccaatat ggccctatgc caatatggct attggccagg ttcaatacta tgtattggcc
     5221 ctatgccata tagtattcca tatatgggtt ttcctattga cgtagatagc ccctcccaat
     5281 gggcggtccc atataccata tatggggctt cctaataccg cccatagcca ctcccccatt
     5341 gacgtcaatg gtctctatat atggtctttc ctattgacgt catatgggcg gtcctattga
     5401 cgtatatggc gcctccccca ttgacgtcaa ttacggtaaa tggcccgcct ggctcaatgc
     5461 ccattgacgt caataggacc acccaccatt gacgtcaatg ggatggctca ttgcccattc
     5521 atatccgttc tcacgccccc tattgacgtc aatgacggta aatggcccac ttggcagtac
     5581 atcaatatct attaatagta acttggcaag tacattacta ttggaagtac gccagggtac
     5641 attggcagta ctcccattga cgtcaatggc ggtaaatggc ccgcgatggc tgccaagtac
     5701 atccccattg acgtcaatgg ggaggggcaa tgacgcaaat gggcgttcca ttgacgtaaa
     5761 tgggcggtag gcgtgcctaa tgggaggtct atataagcaa tgctcgttta gggaac
//
