LOCUS       sequence-453911-        3053 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 0, C terminal tag module (TTCG-GCTT) containing GAL4AD-NLS..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3053)
  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 3053)
  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: pCHW00019
FEATURES             Location/Qualifiers
     source          1..3053
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(194..213)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     313..335
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(373..391)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        459..563
                     /gene="bla"
                     /label=AmpR promoter
     CDS             564..1379
                     /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             564..1379
                     /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     complement(637..656)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1550..2138
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2039..2058
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2292..2309
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2426..2447
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2462..2467,2468..2485,2486..2492)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2500..2516
                     /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)."
     primer_bind     2505..2527
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2524..2540
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2524..2540
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             complement(2612..2632)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(2633..2974)
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="activation domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 activation domain
                     /translation="ANFNQSGNIADSSLSFTFTNSSNGPNLITTQTNSQALSQPIASSN
                     VHDNFMNNEITASKIDDGNNSKPLSPGWTDQTAYNAFGITTGMFNTTTMDDVYNYLFDD
                     EDTPPNPKKE"
     primer_bind     complement(3022..3039)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3022..3038)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3031..3053)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc
       61 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
      121 aatggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatct
      181 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc
      241 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
      301 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
      361 cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
      421 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
      481 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
      541 aataatattg aaaaaggaag agtatgagcc atattcaacg ggaaacgtct tgctctaggc
      601 cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg
      661 tcgggcaatc aggtgcgaca atctatcgat tgtatgggaa gcccgatgcg ccagagttgt
      721 ttctgaaaca tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa
      781 actggctgac ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg
      841 atgcatggtt actcaccact gcgatccctg ggaaaacagc attccaggta ttagaagaat
      901 atcctgattc aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt
      961 cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc
     1021 aatcacgaat gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct
     1081 ggcctgttga acaagtctgg aaagaaatgc ataaactttt gccattctca ccggattcag
     1141 tcgtcactca tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag
     1201 gttgtattga tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat
     1261 ggaactgcct cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta
     1321 ttgataatcc tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaac
     1381 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     1441 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     1501 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     1561 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     1621 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     1681 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     1741 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     1801 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     1861 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     1921 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     1981 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     2041 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     2101 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     2161 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     2221 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     2281 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc
     2341 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
     2401 aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg
     2461 ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc
     2521 acacaggaaa cagctatgac catgctcgag ccaagcttgc atgcaggcct ctgcagtcga
     2581 cgggcccggg atccgatggt ctctaagctt agactttacg cttcttttta ggctcttttt
     2641 ttgggtttgg tggggtatct tcatcatcga atagatagtt atatacatca tccattgtag
     2701 tggtattaaa catccctgta gtgattccaa acgcgttata cgcagtttgg tccgtccaac
     2761 caggtgacag tggttttgaa ttattaccat catcaatttt actagccgtg atttcattat
     2821 tcatgaagtt atcatgaacg ttagaggagg caattggttg tgaaagcgct tgagaatttg
     2881 tttgagttgt tatgaggttc ggaccgttgc tactgttagt gaaagtgaag gacaatgagc
     2941 tatcagcaat attcccactt tgattaaaat tggccgaatg agaccatcta gatgcattcg
     3001 cgaggtaccg agctcgaatt cactggccgt cgttttacaa cgtcgtgact ggg
//
