LOCUS       sequence-452566-        3040 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 0, promoter module (GGAG-CCAT) containing CBS4 and tomato 
            miniDFR promoters..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3040)
  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 3040)
  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: pCHW90004
FEATURES             Location/Qualifiers
     source          1..3040
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(608..1423)
                     /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(608..1423)
                     /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     1331..1350
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(1424..1528)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1596..1614
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1652..1674)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1774..1793
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1987..2009
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2001..2018
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2002..2018
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2487..2503)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2487..2503)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2500..2522)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2511..2527
                     /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(2535..2541,2542..2559,2560..2565))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2580..2601
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2718..2735)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(2889..3040,1..437))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2969..2988)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
       61 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
      121 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
      181 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
      241 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
      301 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
      361 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
      421 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
      481 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
      541 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
      601 ctgacagtta gaaaaactca tcgagcatca aatgaaactg caatttattc atatcaggat
      661 tatcaatacc atatttttga aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc
      721 agttccatag gatggcaaga tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa
      781 tacaacctat taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag
      841 tgacgactga atccggtgag aatggcaaaa gtttatgcat ttctttccag acttgttcaa
      901 caggccagcc attacgctcg tcatcaaaat cactcgcatc aaccaaaccg ttattcattc
      961 gtgattgcgc ctgagcgaga cgaaatacgc gatcgctgtt aaaaggacaa ttacaaacag
     1021 gaatcgaatg caaccggcgc aggaacactg ccagcgcatc aacaatattt tcacctgaat
     1081 caggatattc ttctaatacc tggaatgctg ttttcccagg gatcgcagtg gtgagtaacc
     1141 atgcatcatc aggagtacgg ataaaatgct tgatggtcgg aagaggcata aattccgtca
     1201 gccagtttag tctgaccatc tcatctgtaa catcattggc aacgctacct ttgccatgtt
     1261 tcagaaacaa ctctggcgca tcgggcttcc catacaatcg atagattgtc gcacctgatt
     1321 gcccgacatt atcgcgagcc catttatacc catataaatc agcatccatg ttggaattta
     1381 atcgcggcct agagcaagac gtttcccgtt gaatatggct catactcttc ctttttcaat
     1441 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     1501 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     1561 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     1621 gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg
     1681 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
     1741 gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag
     1801 tgcaccagat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc
     1861 gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
     1921 tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag
     1981 ggttttccca gtcacgacgt tgtaaaacga cggccagtga attcgagctc ggtacctcgc
     2041 gaatgcatct agatggtctc tggagagtgg tcttgggaga gcgatgcgtc ttttccgatg
     2101 aaccgttcca gcaaaaaaga ctaagcgatg cgtcttttcc gctgaaccgt tccagcaaaa
     2161 aagactaagc gatgcgtctt ttccgctgaa ccgttccagc aaaaaagact aggagagcga
     2221 tgcgtctttt ccgctgaacc gttccagcaa aaaagactat cccgtcctaa tacctctacg
     2281 tgcaagcagt ataaaaacaa aatacataaa cataactaaa atctcatatc gttgtaattg
     2341 tcatatacta gtggtaatta agtccagtta cacttaaggt taaatttgct gactcttttt
     2401 tacctttcat ttctgaaacc attgagacca tcggatcccg ggcccgtcga ctgcagaggc
     2461 ctgcatgcaa gcttggctcg agcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     2521 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     2581 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     2641 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     2701 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     2761 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     2821 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     2881 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     2941 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     3001 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
//
