LOCUS       sequence-169834-        6541 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 6541)
  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 6541)
  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-N6(eGFP)
FEATURES             Location/Qualifiers
     source          1..6541
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        18..36
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     18..35
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             join(71..73,74..76,77..787)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(71..73,74..76,77..787)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(116..137)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(377..396)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     724..745
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    830..954
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(991..996,997..1014,1015..1021)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             1075..1734
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1141..1160)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             2076..2381
                     /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     2260..2279
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(2422..2546)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     polyA_signal    2576..2710
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2600..2619)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2654..2673
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2823..2843)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2823..2841)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2862..2878)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2862..2878)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2875..2897)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2886..2902
                     /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(2910..2916,2917..2934,2935..2940))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2955..2976
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3093..3110)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3264..3852)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3344..3363)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4023..4814,4815..4883))
                     /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     4646..4665
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4884..4988)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5014..5469)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5151..5172)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5363..5382
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5540..6524
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 acgtcggagc aagcttgatt taggtgacac tatagaatac aagctacttg ttctttttgc
       61 aggatccacc atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
      121 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga
      181 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
      241 ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga
      301 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
      361 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
      421 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
      481 cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa
      541 gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt
      601 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
      661 cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga
      721 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
      781 gtacaagggc cggccaacaa gtttgtacga gctgtacaag ggccggccaa caagtttgta
      841 caaaaaagct gaacgagaaa cgtaaaatga tataaatatc aatatattaa attagatttt
      901 gcataaaaaa cagactacat aatactgtaa aacacaacat atccagtcac tatggcggcc
      961 cctcgagggg ccgcattagg caccccaggc tttacacttt atgcttccgg ctcgtataat
     1021 gtgtggattt tgagttagga tccgtcgaga ttttcaggag ctaaggaagc taaaatggag
     1081 aaaaaaatca ctggatatac caccgttgat atatcccaat ggcatcgtaa agaacatttt
     1141 gaggcatttc agtcagttgc tcaatgtacc tataaccaga ccgttcagct ggatattacg
     1201 gcctttttaa agaccgtaaa gaaaaataag cacaagtttt atccggcctt tattcacatt
     1261 cttgcccgcc tgatgaatgc tcatccggaa ttccgtatgg caatgaaaga cggtgagctg
     1321 gtgatatggg atagtgttca cccttgttac accgttttcc atgagcaaac tgaaacgttt
     1381 tcatcgctct ggagtgaata ccacgacgat ttccggcagt ttctacacat atattcgcaa
     1441 gatgtggcgt gttacggtga aaacctggcc tatttcccta aagggtttat tgagaatatg
     1501 tttttcgtct cagccaatcc ctgggtgagt ttcaccagtt ttgatttaaa cgtggccaat
     1561 atggacaact tcttcgcccc cgttttcacc atgggcaaat attatacgca aggcgacaag
     1621 gtgctgatgc cgctggcgat tcaggttcat catgccgttt gtgatggctt ccatgtcggc
     1681 agaatgctta atgaattaca acagtactgc gatgagtggc agggcggggc gtaaacgcgt
     1741 ggatccggct tactaaaagc cagataacag tatgcgtatt tgcgcgctga tttttgcggt
     1801 ataagaatat atactgatat gtatacccga agtatgtcaa aaagaggtat gctatgaagc
     1861 agcgtattac agtgacagtt gacagcgaca gctatcagtt gctcaaggca tatatgatgt
     1921 caatatctcc ggtctggtaa gcacaaccat gcagaatgaa gcccgtcgtc tgcgtgccga
     1981 acgctggaaa gcggaaaatc aggaagggat ggctgaggtc gcccggttta ttgaaatgaa
     2041 cggctctttt gctgacgaga acaggggctg gtgaaatgca gtttaaggtt tacacctata
     2101 aaagagagag ccgttatcgt ctgtttgtgg atgtacagag tgatattatt gacacgcccg
     2161 ggcgacggat ggtgatcccc ctggccagtg cacgtctgct gtcagataaa gtctcccgtg
     2221 aactttaccc ggtggtgcat atcggggatg aaagctggcg catgatgacc accgatatgg
     2281 ccagtgtgcc ggtctccgtt atcggggaag aagtggctga tctcagccac cgcgaaaatg
     2341 acatcaaaaa cgccattaac ctgatgttct ggggaatata aatgtcaggc tcccttatac
     2401 acagccagtc tgcaggtcga ccatagtgac tggatatgtt gtgttttaca gtattatgta
     2461 gtctgttttt tatgcaaaat ctaatttaat atattgatat ttatatcatt ttacgtttct
     2521 cgttcagctt tcttgtacaa agtggttcta gaactatagt gagtcgtatt acgtagatcc
     2581 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     2641 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     2701 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg
     2761 ggaggttttt taattcgcgg ccgcggcgcc aatgcattgg gcccggtacc cagcttttgt
     2821 tccctttagt gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg
     2881 tgaaattgtt atccgctcac aattccacac aacatacgag ccgggagcat aaagtgtaaa
     2941 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     3001 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     3061 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     3121 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     3181 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     3241 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     3301 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     3361 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     3421 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     3481 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     3541 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     3601 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     3661 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     3721 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     3781 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     3841 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     3901 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     3961 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     4021 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     4081 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     4141 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     4201 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     4261 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     4321 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     4381 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     4441 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     4501 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     4561 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     4621 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     4681 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     4741 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     4801 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     4861 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     4921 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     4981 gttccgcgca catttccccg aaaagtgcca cctaaattgt aagcgttaat attttgttaa
     5041 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
     5101 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
     5161 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     5221 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
     5281 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     5341 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     5401 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     5461 agggcgcgtc ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
     5521 cctcttcgct attacgccag tcgaccgcca attcaatatg gcgtatatgg actcatgcca
     5581 attcaatatg gtggatctgg acctgtgcca attcaatatg gcgtatatgg actcgtgcca
     5641 attcaatatg gtggatctgg accccagcca attcaatatg gcggacttgg caccatgcca
     5701 attcaatatg gcggacctgg cactgtgcca actggggagg ggtctacttg gcacggtgcc
     5761 aagtttgagg aggggtcttg gccctgtgcc aagtccgcca tattgaattg gcatggtgcc
     5821 aataatggcg gccatattgg ctatatgcca ggatcaatat ataggcaata tccaatatgg
     5881 ccctatgcca atatggctat tggccaggtt caatactatg tattggccct atgccatata
     5941 gtattccata tatgggtttt cctattgacg tagatagccc ctcccaatgg gcggtcccat
     6001 ataccatata tggggcttcc taataccgcc catagccact cccccattga cgtcaatggt
     6061 ctctatatat ggtctttcct attgacgtca tatgggcggt cctattgacg tatatggcgc
     6121 ctcccccatt gacgtcaatt acggtaaatg gcccgcctgg ctcaatgccc attgacgtca
     6181 ataggaccac ccaccattga cgtcaatggg atggctcatt gcccattcat atccgttctc
     6241 acgcccccta ttgacgtcaa tgacggtaaa tggcccactt ggcagtacat caatatctat
     6301 taatagtaac ttggcaagta cattactatt ggaagtacgc cagggtacat tggcagtact
     6361 cccattgacg tcaatggcgg taaatggccc gcgatggctg ccaagtacat ccccattgac
     6421 gtcaatgggg aggggcaatg acgcaaatgg gcgttccatt gacgtaaatg ggcggtaggc
     6481 gtgcctaatg ggaggtctat ataagcaatg ctcgtttagg gaaccgccat tctgcctggg
     6541 g
//
