LOCUS       sequence-169828-        6511 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 6511)
  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 6511)
  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-C6(eGFP)
FEATURES             Location/Qualifiers
     source          1..6511
                     /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             join(1814..1816,1817..1819,1820..2533)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(1859..1880)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2120..2139)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2467..2488
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2563..2697
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2587..2606)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2641..2660
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2810..2830)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2810..2828)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2849..2865)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2849..2865)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2862..2884)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2873..2889
                     /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(2897..2903,2904..2921,2922..2927))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2942..2963
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3080..3097)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3251..3839)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3331..3350)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4010..4801,4802..4870))
                     /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     4633..4652
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4871..4975)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5001..5456)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5138..5159)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5350..5369
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5527..6511
                     /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 atcatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
     1861 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg
     1921 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
     1981 gccctggccc accctcgtga ccaccttcac ctacggcgtg cagtgcttca gccgctaccc
     2041 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
     2101 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
     2161 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
     2221 catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga
     2281 caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag
     2341 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     2401 gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg
     2461 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactcacg gcatggacga
     2521 gctgtacaag taatctagaa ctatagtgag tcgtattacg tagatccaga catgataaga
     2581 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
     2641 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
     2701 aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa
     2761 ttcgcggccg cggcgccaat gcattgggcc cggtacccag cttttgttcc ctttagtgag
     2821 ggttaattgc gcgcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     2881 cgctcacaat tccacacaac atacgagccg ggagcataaa gtgtaaagcc tggggtgcct
     2941 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     3001 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     3061 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     3121 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     3181 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     3241 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     3301 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     3361 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     3421 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     3481 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     3541 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     3601 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     3661 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     3721 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     3781 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     3841 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     3901 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     3961 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     4021 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     4081 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     4141 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     4201 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     4261 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     4321 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     4381 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     4441 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     4501 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     4561 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     4621 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     4681 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     4741 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     4801 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     4861 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     4921 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     4981 ttccccgaaa agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa
     5041 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
     5101 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
     5161 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     5221 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     5281 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     5341 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
     5401 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca
     5461 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
     5521 acgccagtcg accgccaatt caatatggcg tatatggact catgccaatt caatatggtg
     5581 gatctggacc tgtgccaatt caatatggcg tatatggact cgtgccaatt caatatggtg
     5641 gatctggacc ccagccaatt caatatggcg gacttggcac catgccaatt caatatggcg
     5701 gacctggcac tgtgccaact ggggaggggt ctacttggca cggtgccaag tttgaggagg
     5761 ggtcttggcc ctgtgccaag tccgccatat tgaattggca tggtgccaat aatggcggcc
     5821 atattggcta tatgccagga tcaatatata ggcaatatcc aatatggccc tatgccaata
     5881 tggctattgg ccaggttcaa tactatgtat tggccctatg ccatatagta ttccatatat
     5941 gggttttcct attgacgtag atagcccctc ccaatgggcg gtcccatata ccatatatgg
     6001 ggcttcctaa taccgcccat agccactccc ccattgacgt caatggtctc tatatatggt
     6061 ctttcctatt gacgtcatat gggcggtcct attgacgtat atggcgcctc ccccattgac
     6121 gtcaattacg gtaaatggcc cgcctggctc aatgcccatt gacgtcaata ggaccaccca
     6181 ccattgacgt caatgggatg gctcattgcc cattcatatc cgttctcacg ccccctattg
     6241 acgtcaatga cggtaaatgg cccacttggc agtacatcaa tatctattaa tagtaacttg
     6301 gcaagtacat tactattgga agtacgccag ggtacattgg cagtactccc attgacgtca
     6361 atggcggtaa atggcccgcg atggctgcca agtacatccc cattgacgtc aatggggagg
     6421 ggcaatgacg caaatgggcg ttccattgac gtaaatgggc ggtaggcgtg cctaatggga
     6481 ggtctatata agcaatgctc gtttagggaa c
//
