LOCUS       sequence-169836-        5882 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 5882)
  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 5882)
  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-C3m(3xFLAG)
FEATURES             Location/Qualifiers
     source          1..5882
                     /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"
     CDS             1829..1852
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1856..1879
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1883..1906
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    1934..2068
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1958..1977)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2012..2031
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2181..2201)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2181..2199)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2220..2236)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2220..2236)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2233..2255)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2244..2260
                     /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(2268..2274,2275..2292,2293..2298))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2313..2334
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2451..2468)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2622..3210)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2702..2721)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3381..4172,4173..4241))
                     /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     4004..4023
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4242..4346)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4372..4827)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4509..4530)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4721..4740
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4898..5882
                     /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 aagacaggat ccaccatgga ttataaggat gacgatgaca aagcagacta
     1861 caaagacgac gatgataagg ctgattataa agatgatgac gacaaataga actatagtga
     1921 gtcgtattac gtagatccag acatgataag atacattgat gagtttggac aaaccacaac
     1981 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
     2041 aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca
     2101 ggttcagggg gaggtgtggg aggtttttta attcgcggcc gcggcgccaa tgcattgggc
     2161 ccggtaccca gcttttgttc cctttagtga gggttaattg cgcgcttggc gtaatcatgg
     2221 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     2281 gggagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     2341 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     2401 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     2461 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     2521 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     2581 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     2641 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     2701 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     2761 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     2821 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     2881 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     2941 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     3001 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     3061 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     3121 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     3181 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     3241 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     3301 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     3361 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     3421 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     3481 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     3541 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     3601 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     3661 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     3721 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     3781 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     3841 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     3901 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     3961 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     4021 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     4081 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     4141 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     4201 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     4261 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     4321 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc taaattgtaa
     4381 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     4441 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     4501 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     4561 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     4621 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     4681 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     4741 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     4801 cgcttaatgc gccgctacag ggcgcgtccc attcgccatt caggctgcgc aactgttggg
     4861 aagggcgatc ggtgcgggcc tcttcgctat tacgccagtc gaccgccaat tcaatatggc
     4921 gtatatggac tcatgccaat tcaatatggt ggatctggac ccgtgccaat tcaatatggc
     4981 gtatatggac tcgtgccaat tcaatatggt ggatctggac cccagccaat tcaatatggc
     5041 ggacttggca ccatgccaat tcaatatggc ggacctggca ctgtgccaac tggggagggg
     5101 tctacttggc acggtgccaa gtttgaggag gggtcttggc cctgtgccaa gtccgccata
     5161 ttgaattggc atggtgccaa taatggcggc catattggct atatgccagg atcaatatat
     5221 aggcaatatc caatatggcc ctatgccaat atggctattg gccaggttca atactatgta
     5281 ttggccctat gccatatagt attccatata tgggttttcc tattgacgta gatagcccct
     5341 cccaatgggc ggtcccatat accatatatg gggcttccta ataccgccca tagccactcc
     5401 cccattgacg tcaatggtct ctatatatgg tctttcctat tgacgtcata tgggcggtcc
     5461 tattgacgta tatggcgcct cccccattga cgtcaattac ggtaaatggc ccgcctggct
     5521 caatgcccat tgacgtcaat aggaccaccc accattgacg tcaatgggat ggctcattgc
     5581 ccattcatat ccgttctcac gccccctatt gacgtcaatg acggtaaatg gcccacttgg
     5641 cagtacatca atatctatta atagtaactt ggcaagtaca ttactattgg aagtacgcca
     5701 gggtacattg gcagtactcc cattgacgtc aatggcggta aatggcccgc gatggctgcc
     5761 aagtacatcc ccattgacgt caatggggag gggcaatgac gcaaatgggc gttccattga
     5821 cgtaaatggg cggtaggcgt gcctaatggg aggtctatat aagcaatgct cgtttaggga
     5881 ac
//
