LOCUS       sequence-176309-        5872 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 5872)
  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 5872)
  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-N2(3xHA)
FEATURES             Location/Qualifiers
     source          1..5872
                     /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             88..168
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     protein_bind    178..302
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(339..344,345..362,363..369)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             423..1082
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(489..508)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             1424..1729
                     /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     1608..1627
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(1770..1894)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     polyA_signal    1924..2058
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1948..1967)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2002..2021
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2171..2191)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2171..2189)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2210..2226)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2210..2226)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2223..2245)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2234..2250
                     /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(2258..2264,2265..2282,2283..2288))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2303..2324
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2441..2458)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2612..3200)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2692..2711)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3371..4162,4163..4231))
                     /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     3994..4013
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4232..4336)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4362..4817)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4499..4520)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4711..4730
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4888..5872
                     /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 atccatgtac ccatacgatg ttccagatta cgcttatcct
      121 tatgacgtac ctgactatgc atacccttat gatgtaccag actacgctgg ccggccaaca
      181 agtttgtaca aaaaagctga acgagaaacg taaaatgata taaatatcaa tatattaaat
      241 tagattttgc ataaaaaaca gactacataa tactgtaaaa cacaacatat ccagtcacta
      301 tggcggcccc tcgaggggcc gcattaggca ccccaggctt tacactttat gcttccggct
      361 cgtataatgt gtggattttg agttaggatc cgtcgagatt ttcaggagct aaggaagcta
      421 aaatggagaa aaaaatcact ggatatacca ccgttgatat atcccaatgg catcgtaaag
      481 aacattttga ggcatttcag tcagttgctc aatgtaccta taaccagacc gttcagctgg
      541 atattacggc ctttttaaag accgtaaaga aaaataagca caagttttat ccggccttta
      601 ttcacattct tgcccgcctg atgaatgctc atccggaatt ccgtatggca atgaaagacg
      661 gtgagctggt gatatgggat agtgttcacc cttgttacac cgttttccat gagcaaactg
      721 aaacgttttc atcgctctgg agtgaatacc acgacgattt ccggcagttt ctacacatat
      781 attcgcaaga tgtggcgtgt tacggtgaaa acctggccta tttccctaaa gggtttattg
      841 agaatatgtt tttcgtctca gccaatccct gggtgagttt caccagtttt gatttaaacg
      901 tggccaatat ggacaacttc ttcgcccccg ttttcaccat gggcaaatat tatacgcaag
      961 gcgacaaggt gctgatgccg ctggcgattc aggttcatca tgccgtttgt gatggcttcc
     1021 atgtcggcag aatgcttaat gaattacaac agtactgcga tgagtggcag ggcggggcgt
     1081 aaacgcgtgg atccggctta ctaaaagcca gataacagta tgcgtatttg cgcgctgatt
     1141 tttgcggtat aagaatatat actgatatgt atacccgaag tatgtcaaaa agaggtatgc
     1201 tatgaagcag cgtattacag tgacagttga cagcgacagc tatcagttgc tcaaggcata
     1261 tatgatgtca atatctccgg tctggtaagc acaaccatgc agaatgaagc ccgtcgtctg
     1321 cgtgccgaac gctggaaagc ggaaaatcag gaagggatgg ctgaggtcgc ccggtttatt
     1381 gaaatgaacg gctcttttgc tgacgagaac aggggctggt gaaatgcagt ttaaggttta
     1441 cacctataaa agagagagcc gttatcgtct gtttgtggat gtacagagtg atattattga
     1501 cacgcccggg cgacggatgg tgatccccct ggccagtgca cgtctgctgt cagataaagt
     1561 ctcccgtgaa ctttacccgg tggtgcatat cggggatgaa agctggcgca tgatgaccac
     1621 cgatatggcc agtgtgccgg tctccgttat cggggaagaa gtggctgatc tcagccaccg
     1681 cgaaaatgac atcaaaaacg ccattaacct gatgttctgg ggaatataaa tgtcaggctc
     1741 ccttatacac agccagtctg caggtcgacc atagtgactg gatatgttgt gttttacagt
     1801 attatgtagt ctgtttttta tgcaaaatct aatttaatat attgatattt atatcatttt
     1861 acgtttctcg ttcagctttc ttgtacaaag tggttctaga actatagtga gtcgtattac
     1921 gtagatccag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     1981 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     2041 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
     2101 gaggtgtggg aggtttttta attcgcggcc gcggcgccaa tgcattgggc ccggtaccca
     2161 gcttttgttc cctttagtga gggttaattg cgcgcttggc gtaatcatgg tcatagctgt
     2221 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc gggagcataa
     2281 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     2341 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     2401 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     2461 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     2521 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     2581 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     2641 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     2701 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     2761 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     2821 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     2881 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     2941 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     3001 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     3061 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     3121 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     3181 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     3241 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     3301 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     3361 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     3421 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     3481 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     3541 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     3601 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     3661 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     3721 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     3781 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     3841 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     3901 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     3961 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     4021 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     4081 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     4141 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     4201 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     4261 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     4321 aaataggggt tccgcgcaca tttccccgaa aagtgccacc taaattgtaa gcgttaatat
     4381 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     4441 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     4501 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     4561 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     4621 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     4681 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     4741 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     4801 gccgctacag ggcgcgtccc attcgccatt caggctgcgc aactgttggg aagggcgatc
     4861 ggtgcgggcc tcttcgctat tacgccagtc gaccgccaat tcaatatggc gtatatggac
     4921 tcatgccaat tcaatatggt ggatctggac ctgtgccaat tcaatatggc gtatatggac
     4981 tcgtgccaat tcaatatggt ggatctggac cccagccaat tcaatatggc ggacttggca
     5041 ccatgccaat tcaatatggc ggacctggca ctgtgccaac tggggagggg tctacttggc
     5101 acggtgccaa gtttgaggag gggtcttggc cctgtgccaa gtccgccata ttgaattggc
     5161 atggtgccaa taatggcggc catattggct atatgccagg atcaatatat aggcaatatc
     5221 caatatggcc ctatgccaat atggctattg gccaggttca atactatgta ttggccctat
     5281 gccatatagt attccatata tgggttttcc tattgacgta gatagcccct cccaatgggc
     5341 ggtcccatat accatatatg gggcttccta ataccgccca tagccactcc cccattgacg
     5401 tcaatggtct ctatatatgg tctttcctat tgacgtcata tgggcggtcc tattgacgta
     5461 tatggcgcct cccccattga cgtcaattac ggtaaatggc ccgcctggct caatgcccat
     5521 tgacgtcaat aggaccaccc accattgacg tcaatgggat ggctcattgc ccattcatat
     5581 ccgttctcac gccccctatt gacgtcaatg acggtaaatg gcccacttgg cagtacatca
     5641 atatctatta atagtaactt ggcaagtaca ttactattgg aagtacgcca gggtacattg
     5701 gcagtactcc cattgacgtc aatggcggta aatggcccgc gatggctgcc aagtacatcc
     5761 ccattgacgt caatggggag gggcaatgac gcaaatgggc gttccattga cgtaaatggg
     5821 cggtaggcgt gcctaatggg aggtctatat aagcaatgct cgtttaggga ac
//
