LOCUS       sequence-169868-        6026 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 6026)
  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 6026)
  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-C1m(5xMYC)
FEATURES             Location/Qualifiers
     source          1..6026
                     /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             1841..1870
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1880..1909
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1919..1948
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1958..1987
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1997..2026
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     polyA_signal    2078..2212
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2102..2121)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2156..2175
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2325..2345)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2325..2343)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2364..2380)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2364..2380)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2377..2399)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2388..2404
                     /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(2412..2418,2419..2436,2437..2442))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2457..2478
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2595..2612)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2766..3354)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2846..2865)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3525..4316,4317..4385))
                     /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     4148..4167
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4386..4490)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4516..4971)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4653..4674)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4865..4884
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5042..6026
                     /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 cccatcgatt taaagctatg gagcaaaagc tcatttctga
     1861 agaggacttg aatgaaatgg agcaaaagct catttctgaa gaggacttga atgaaatgga
     1921 gcaaaagctc atttctgaag aggacttgaa tgaaatggag caaaagctca tttctgaaga
     1981 ggacttgaat gaaatggagc aaaagctcat ttctgaagag gacttgaatg aaatggagag
     2041 cttgggagac tagaactata gtgagtcgta ttacgtagat ccagacatga taagatacat
     2101 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     2161 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
     2221 caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaattcgc
     2281 ggccgcggcg ccaatgcatt gggcccggta cccagctttt gttcccttta gtgagggtta
     2341 attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     2401 acaattccac acaacatacg agccgggagc ataaagtgta aagcctgggg tgcctaatga
     2461 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     2521 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     2581 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     2641 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     2701 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     2761 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     2821 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     2881 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     2941 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     3001 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     3061 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     3121 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     3181 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     3241 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     3301 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     3361 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     3421 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     3481 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     3541 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     3601 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     3661 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     3721 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     3781 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     3841 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     3901 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     3961 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     4021 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     4081 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     4141 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     4201 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     4261 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     4321 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     4381 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     4441 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     4501 cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     4561 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     4621 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     4681 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     4741 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     4801 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     4861 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     4921 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc
     4981 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
     5041 agtcgaccgc caattcaata tggcgtatat ggactcatgc caattcaata tggtggatct
     5101 ggacctgtgc caattcaata tggcgtatat ggactcgtgc caattcaata tggtggatct
     5161 ggaccccagc caattcaata tggcggactt ggcaccatgc caattcaata tggcggacct
     5221 ggcactgtgc caactgggga ggggtctact tggcacggtg ccaagtttga ggaggggtct
     5281 tggccctgtg ccaagtccgc catattgaat tggcatggtg ccaataatgg cggccatatt
     5341 ggctatatgc caggatcaat atataggcaa tatccaatat ggccctatgc caatatggct
     5401 attggccagg ttcaatacta tgtattggcc ctatgccata tagtattcca tatatgggtt
     5461 ttcctattga cgtagatagc ccctcccaat gggcggtccc atataccata tatggggctt
     5521 cctaataccg cccatagcca ctcccccatt gacgtcaatg gtctctatat atggtctttc
     5581 ctattgacgt catatgggcg gtcctattga cgtatatggc gcctccccca ttgacgtcaa
     5641 ttacggtaaa tggcccgcct ggctcaatgc ccattgacgt caataggacc acccaccatt
     5701 gacgtcaatg ggatggctca ttgcccattc atatccgttc tcacgccccc tattgacgtc
     5761 aatgacggta aatggcccac ttggcagtac atcaatatct attaatagta acttggcaag
     5821 tacattacta ttggaagtac gccagggtac attggcagta ctcccattga cgtcaatggc
     5881 ggtaaatggc ccgcgatggc tgccaagtac atccccattg acgtcaatgg ggaggggcaa
     5941 tgacgcaaat gggcgttcca ttgacgtaaa tgggcggtag gcgtgcctaa tgggaggtct
     6001 atataagcaa tgctcgttta gggaac
//
