LOCUS       sequence-454056-        3083 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 0, C terminal tag module (TTCG-GCTT) containing p65AD-NLS..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3083)
  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 3083)
  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: pCHW00016
FEATURES             Location/Qualifiers
     source          1..3083
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      632..1220
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1121..1140
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1374..1391
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1508..1529
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1544..1549,1550..1567,1568..1574)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1582..1598
                     /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     1587..1609
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1606..1622
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1606..1622
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             complement(1694..1714)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(1715..2086)
                     /codon_start=1
                     /product="transcriptional activation domain of human RelA, 
                     also known as p65 (O'Shea and Perkins, 2008)"
                     /label=RelA (p65) AD
                     /translation="PTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNS
                     EFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGD
                     EDFSSIADMDFSALLSQISS"
     primer_bind     complement(2134..2151)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2134..2150)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2143..2165)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2359..2378)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2478..2500
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2538..2556)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2624..2728
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2729..3083,1..461)
                     /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             join(2729..3083,1..461)
                     /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(2802..2821)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 atattgttga tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt
       61 gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga atgaataacg
      121 gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct
      181 ggaaagaaat gcataaactt ttgccattct caccggattc agtcgtcact catggtgatt
      241 tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac
      301 gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt
      361 tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga
      421 ataaattgca gtttcatttg atgctcgatg agtttttcta actgtcagac caagtttact
      481 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
      541 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
      601 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
      661 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
      721 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
      781 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
      841 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
      901 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
      961 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     1021 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     1081 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     1141 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     1201 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     1261 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     1321 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     1381 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     1441 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     1501 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     1561 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     1621 accatgctcg agccaagctt gcatgcaggc ctctgcagtc gacgggcccg ggatccgatg
     1681 gtctctaagc ttagacttta cgcttctttt taggggagct tatctggctc agcaaagcgg
     1741 agaaatccat atcagcgatg ctggagaaat cctcatcgcc gctgaggagg ccattgggca
     1801 aacccggggc acccagtggg gcaggggctg gatcgggtgg tcgctgtgcg ccggtcacca
     1861 gtcgcgtgat agcctccggg tattccatca gcatgggctc tgtggtatga ggtgcgaccg
     1921 ggattccctg attcaagagc tgctgaaact cgctgttgtc cacggatgcg agatcggtga
     1981 atacggcagg gtctgtgctg ttacccagca gggctcccaa atcttcgtca tcgaactgaa
     2041 gctgcaagag ggcctcgctc agggttcctt cgcccgcctg cgttggcgaa tgagaccatc
     2101 tagatgcatt cgcgaggtac cgagctcgaa ttcactggcc gtcgttttac aacgtcgtga
     2161 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
     2221 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
     2281 tggcgaatgg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     2341 catctggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca
     2401 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
     2461 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
     2521 acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     2581 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     2641 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     2701 gcttcaataa tattgaaaaa ggaagagtat gagccatatt caacgggaaa cgtcttgctc
     2761 taggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga
     2821 taatgtcggg caatcaggtg cgacaatcta tcgattgtat gggaagcccg atgcgccaga
     2881 gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag
     2941 actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc
     3001 tgatgatgca tggttactca ccactgcgat ccctgggaaa acagcattcc aggtattaga
     3061 agaatatcct gattcaggtg aaa
//
