LOCUS       sequence-454078-        3364 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 0, CDSns module (AATG-TTCG) containing S. cerevisiae CUP2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3364)
  AUTHORS   Chiang BJ, Lin KY, Chen YF, Huang CY, Goh FJ, Huang LT, Chen LH, Wu 
            CH
  TITLE     Development of a tightly regulated copper-inducible transient gene 
            expression system in Nicotiana benthamiana incorporating suicide 
            exon and Cre recombinase
  JOURNAL   bioRxiv 2024.03.23.586378
REFERENCE   2  (bases 1 to 3364)
  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: "bioRxiv 
            2024.03.23.586378"
COMMENT     Sequence Label: pCHW00011
FEATURES             Location/Qualifiers
     source          1..3364
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1188..1204)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1188..1204)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1201..1223)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1212..1228
                     /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(1236..1242,1243..1260,1261..1266))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1281..1302
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1419..1436)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1590..2178)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1670..1689)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2349..3164)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(2349..3164)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     3072..3091
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(3165..3269)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3337..3355
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accagatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cgagctcggt acctcgcgaa
      421 tgcatctaga tggtctctaa tggtcgtaat taacggggtc aaatatgcct gtgaaacgtg
      481 tatcaggggt cacagggcgg cgcagtgtac tcacactgat ggtccgctac agatgatcag
      541 acgcaagggt agaccatcga ccacatgtgg ccattgtaaa gagctgagaa gaaccaagaa
      601 cttcaaccca tccggtgggt gcatgtgtgc ctctgcacga cggccagctg ttggcagcaa
      661 ggaagatgaa acacgatgtc gttgtgatga gggtgaacct tgtaaatgtc ataccaagag
      721 gaaaagcagc cggaaatcaa agggagggtc atgccacaga agggcaaatg atgaagcagc
      781 gcatgtcaat ggtcttggta ttgcagatct ggacgttctt ttgggcctaa atggtcgctc
      841 gtcggatgta gacatgacaa ccacattgcc gagtttgaag ccacctctgc aaaacggaga
      901 aattaaggcc gacagcattg acaatcttga tttggcttcc ctcgatccgc ttgagcaaag
      961 ccctagtata tctatggaac ctgttagtat caatgaaaca ggaagcgcat atacaactac
     1021 gaacacagca ctaaacgata ttgacattcc attctccatc aatgagttga acgagctata
     1081 caaacaagta tcttcgcata actcacattc acagccatat tcgtgagacc atcggatccc
     1141 gggcccgtcg actgcagagg cctgcatgca agcttggctc gagcatggtc atagctgttt
     1201 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     1261 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     1321 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     1381 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     1441 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     1501 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     1561 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     1621 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     1681 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     1741 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     1801 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     1861 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     1921 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     1981 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     2041 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     2101 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     2161 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     2221 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     2281 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     2341 tctgacagtt agaaaaactc atcgagcatc aaatgaaact gcaatttatt catatcagga
     2401 ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg
     2461 cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg tccaacatca
     2521 atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa atcaccatga
     2581 gtgacgactg aatccggtga gaatggcaaa agtttatgca tttctttcca gacttgttca
     2641 acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc gttattcatt
     2701 cgtgattgcg cctgagcgag acgaaatacg cgatcgctgt taaaaggaca attacaaaca
     2761 ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt ttcacctgaa
     2821 tcaggatatt cttctaatac ctggaatgct gttttcccag ggatcgcagt ggtgagtaac
     2881 catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat aaattccgtc
     2941 agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc tttgccatgt
     3001 ttcagaaaca actctggcgc atcgggcttc ccatacaatc gatagattgt cgcacctgat
     3061 tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat gttggaattt
     3121 aatcgcggcc tagagcaaga cgtttcccgt tgaatatggc tcatactctt cctttttcaa
     3181 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     3241 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     3301 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt
     3361 cgtc
//
