LOCUS       sequence-472306-        9347 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 1, Position 3 containing CBS4-CMV::RUBY::OCS..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9347)
  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 9347)
  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: pCHW111040
FEATURES             Location/Qualifiers
     source          1..9347
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    42..66
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(222..1370)
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     rep_origin      complement(1706..2294)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1786..1805)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2465..3256,3257..3325))
                     /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     3088..3107
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3326..3430)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3517..4148
                     /label=oriV
                     /note="incP origin of replication"
     misc_feature    4240..4264
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             4669..8619
                     /label=RUBY
     CDS             6169..6225
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             7060..7116
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     terminator      8633..9340
                     /label=OCS terminator
                     /note="octopine synthase terminator"
ORIGIN
        1 ttacttgtct tctgcacgaa gtggtttaaa ctatcagtgt ttgacaggat atattggcgg
       61 gtaaacctaa gagaaaagag cgtttattag aataatcgga tatttaaaag ggcgtgaaaa
      121 ggtttatccg ttcgtccatt tgtatgtgca tgccaaccac agggttcccc agatcaggcg
      181 ctggctgctg aacccccagc cggaactgac cccacaaggc cctagcgttt gcaatgcacc
      241 aggtcatcat tgacccaggc gtgttccacc aggccgctgc ctcgcaactc ttcgcaggct
      301 tcgccgacct gctcgcgcca cttcttcacg cgggtggaat ccgatccgca catgaggcgg
      361 aaggtttcca gcttgagcgg gtacggctcc cggtgcgagc tgaaatagtc gaacatccgt
      421 cgggccgtcg gcgacagctt gcggtacttc tcccatatga atttcgtgta gtggtcgcca
      481 gcaaacagca cgacgatttc ctcgtcgatc aggacctggc aacgggacgt tttcttgcca
      541 cggtccagga cgcggaagcg gtgcagcagc gacaccgatt ccaggtgccc aacgcggtcg
      601 gacgtgaagc ccatcgccgt cgcctgtagg cgcgacaggc attcctcggc cttcgtgtaa
      661 taccggccat tgatcgacca gcccaggtcc tggcaaagct cgtagaacgt gaaggtgatc
      721 ggctcgccga taggggtgcg cttcgcgtac tccaacacct gctgccacac cagttcgtca
      781 tcgtcggccc gcagctcgac gccggtgtag gtgatcttca cgtccttgtt gacgtggaaa
      841 atgaccttgt tttgcagcgc ctcgcgcggg attttcttgt tgcgcgtggt gaacagggca
      901 gagcgggccg tgtcgtttgg catcgctcgc atcgtgtccg gccacggcgc aatatcgaac
      961 aaggaaagct gcatttcctt gatctgctgc ttcgtgtgtt tcagcaacgc ggcctgcttg
     1021 gcctcgctga cctgttttgc caggtcctcg ccggcggttt ttcgcttctt ggtcgtcata
     1081 gttcctcgcg tgtcgatggt catcgacttc gccaaacctg ccgcctcctg ttcaagacga
     1141 cgcgaacgct ccacggcggc cgatggcgcg ggcagggcag ggggagccag ttgcacgctg
     1201 tcgcgctcga tcttggccgt agcttgctgg accatcgagc cgacggactg gaaggtttcg
     1261 cggggcgcac gcatgacggt gcggcttgcg atggtttcgg catcctcggc ggaaaacccc
     1321 gcgtcgatca gttcttgcct gtatgccttc cggtcaaacg tccgattcat tcaccctcct
     1381 tgcgggattg ccccgactca cgccggggca atgtgccctt attcctgatt tgacccgcct
     1441 ggtgccttgg tgtccagata atccacctta tcggcaatga agtcggtccc gtagaccgtc
     1501 tggccgtcct tctcgtactt ggtattccga atcttgccct gcacgaatac cagcgacccc
     1561 ttgcccaaat acttgccgtg ggcctcggcc tgagagccaa aacacttgat gcggaagaag
     1621 tcggtgcgct cctgcttgtc gccggcatcg ttgcgccaca tctaggatct gccaggaacc
     1681 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     1741 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     1801 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     1861 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     1921 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     1981 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     2041 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     2101 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     2161 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     2221 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     2281 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     2341 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     2401 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     2461 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     2521 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     2581 gccccagtgc tgcaatgata ccgcgagaac cacgctcacc ggctccagat ttatcagcaa
     2641 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     2701 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     2761 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     2821 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     2881 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     2941 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     3001 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     3061 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     3121 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     3181 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     3241 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     3301 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     3361 aaggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     3421 gggttccgcg cacgaattgg ccagcgctgc catttttggg gtgaggccgt tcgcggccga
     3481 ggggcgcagc ccctgggggg atgggaggcc cgcgttagcg ggccgggagg gttcgagaag
     3541 ggggggcacc ccccttcggc gtgcgcggtc acgcgcacag ggcgcagccc tggttaaaaa
     3601 caaggtttat aaatattggt ttaaaagcag gttaaaagac aggttagcgg tggccgaaaa
     3661 acgggcggaa acccttgcaa atgctggatt ttctgcctgt ggacagcccc tcaaatgtca
     3721 ataggtgcgc ccctcatctg tcagcactct gcccctcaag tgtcaaggat cgcgcccctc
     3781 atctgtcagt agtcgcgccc ctcaagtgtc aataccgcag ggcacttatc cccaggcttg
     3841 tccacatcat ctgtgggaaa ctcgcgtaaa atcaggcgtt ttcgccgatt tgcgaggctg
     3901 gccagctcca cgtcgccggc cgaaatcgag cctgcccctc atctgtcaac gccgcgccgg
     3961 gtgagtcggc ccctcaagtg tcaacgtccg cccctcatct gtcagtgagg gccaagtttt
     4021 ccgcgaggta tccacaacgc cggcggccgc ggtgtctcgc acacggcttc gacggcgttt
     4081 ctggcgcgtt tgcagggcca tagacggccg ccagcccagc ggcgagggca accagcccgg
     4141 tgagcgtcgc aaaggagatc ctgatctgac tgatgggctg cctgtatcga gtggtgattt
     4201 tgtgccgagc tgccggtcgg ggagctgttg gctggctggt ggcaggatat attgtggtgt
     4261 aaacaaattg acgcttagac aacttaataa cacattgcgg acgtttttaa tgtactgggg
     4321 tggatgcagt gggccccact ctgtgaagac aaactagaat tcgagctcgg agagtggtct
     4381 tgggagagcg atgcgtcttt tccgatgaac cgttccagca aaaaagacta agcgatgcgt
     4441 cttttccgct gaaccgttcc agcaaaaaag actaagcgat gcgtcttttc cgctgaaccg
     4501 ttccagcaaa aaagactagg agagcgatgc gtcttttccg ctgaaccgtt ccagcaaaaa
     4561 agactatccc tactgtttat ttacaagagc gtacggttca atccctgcct cccctgtaaa
     4621 actacccttt gaaaacctct ctttcttaat cttttctttg taattccaat ggatcatgcg
     4681 accctcgcca tgatcctcgc gatctggttc atcagcttcc acttcatcaa gctgctgttc
     4741 tcccagcaga ccaccaagct gcttccgcca ggaccaaagc cgcttccgat catcggcaac
     4801 atccttgagg tgggcaagaa gccgcatcgg tccttcgcca acctcgccaa gattcacggc
     4861 ccactcattt ccctcagact cggctctgtg accaccatcg ttgtgtcctc tgccgacgtg
     4921 gccaaagaga tgttcctcaa gaaggatcac ccgctctcca accgcacgat cccgaatagt
     4981 gttacagccg gcgaccacca caagctcacc atgtcttggc tcccggtgtc tccgaagtgg
     5041 cgcaacttcc gcaagattac cgccgtgcat ctgctctccc cacagagact cgatgcctgc
     5101 cagacattca ggcacgccaa ggtgcagcag ctctacgagt acgttcaaga gtgcgcccag
     5161 aaaggccagg ccgtggatat tggcaaggcc gcctttacga ccagcctcaa cctcctcagc
     5221 aagctgttct tcagcgtcga gctggcgcac cacaagtccc ataccagcca agagttcaaa
     5281 gagctgatct ggaacatcat ggaagatata ggcaagccga actacgccga ctacttcccg
     5341 attctcggct gcgttgaccc atctggcatt agaagaaggc tcgcctgctc cttcgacaag
     5401 ctgatcgccg tgttccaggg catcatctgc gagagactcg ccccagattc ctccaccaca
     5461 actaccacca ccaccgacga cgtgctcgat gtgctcctcc agctgttcaa gcagaacgag
     5521 ctgacgatgg gcgagatcaa ccacctcctc gtggacatct tcgacgccgg caccgatacc
     5581 acatcctcca cattcgagtg ggtgatgacc gagctgatcc gcaatccaga gatgatggaa
     5641 aaggcccaag aggaaatcaa gcaggtcctc ggcaaggaca agcagatcca agagtccgac
     5701 atcatcaacc tgccgtacct ccaggcgatc atcaaagaga cactccgcct ccatccgccg
     5761 accgtgttct tgctcccaag aaaggccgac accgatgtcg agctgtacgg ctacatcgtg
     5821 ccgaaggatg cccagatcct cgtgaacctc tgggccattg gcagggaccc aaacgcctgg
     5881 cagaacgccg atattttcag cccagagcgc ttcatcggct gcgagatcga tgttaagggc
     5941 cgcgatttcg gcctccttcc atttggcgct ggccgcagaa tttgcccagg catgaatctc
     6001 gccatcagga tgctcaccct catgctcgcc acactcctcc agttcttcaa ctggaagctc
     6061 gaaggcgaca tctccccgaa ggacctcgac atggacgaga agttcggcat tgcgctccaa
     6121 aagaccaagc cgctcaagct catcccgatt ccgcgctacg gtagcggagc taccaatttt
     6181 agcctcctta agcaggcagg tgatgtagaa gagaaccccg ggcctatgaa gatgatgaac
     6241 ggcgaggacg ccaacgacca gatgatcaaa gagtccttct tcatcaccca cggcaacccg
     6301 atcctcaccg tcgaggatac acatccgctc aggccgttct tcgagacatg gcgcgagaag
     6361 attttctcca agaagccgaa ggccatcctc atcatctccg gccactggga gacagtgaag
     6421 ccaaccgtga acgccgtgca catcaacgac accatccacg acttcgacga ctacccagcc
     6481 gccatgtacc agttcaagta cccagctcca ggcgagccag agcttgcgag aaaggtggaa
     6541 gagatcctca agaagtccgg gttcgagaca gccgagacag accaaaagag gggccttgat
     6601 cacggcgcct gggttccact catgctcatg tatccagagg cggacatccc ggtgtgccag
     6661 ctctcagttc agccacatct cgacggcacc taccactaca atctcggcag agccctcgcg
     6721 ccgctcaaga atgatggcgt gctcattatt ggctccggca gcgccacaca tccactcgat
     6781 gagacaccgc actacttcga tggtgttgcc ccttgggccg ctgccttcga ttcttggctt
     6841 aggaaggccc tcatcaacgg ccgcttcgag gaagtgaaca tctacgagag caaggccccg
     6901 aactggaagc tcgcccatcc atttccagag cacttctacc cgctccacgt tgtgctcggc
     6961 gctgctggtg aaaagtggaa ggccgagctg atccactcct cctgggatca tggcacactt
     7021 tgccacggct cctacaagtt cacctccgcc ggatccggag caaccaactt tagcctgctc
     7081 aagcaagcag gagatgttga ggaaaatcct ggccccatga ccgccatcaa gatgaacacc
     7141 aacggcgagg gcgagacaca gcacatcctc atgatcccgt tcatggcgca gggccacctc
     7201 aggccatttc tcgaactcgc catgttcctc tacaagcgct cccacgtgat catcaccctg
     7261 ctcacaactc cgctcaatgc cggcttcctc aggcacctcc ttcaccacca ttcctactcc
     7321 tccagcggca tcaggatcgt cgagctgcca ttcaactcca ccaaccacgg actcccaccg
     7381 ggcatcgaga acaccgataa gctcacactc ccgctcgtgg tgtccctctt ccattccacc
     7441 atcagcctcg atccgcacct ccgcgattac atctccaggc atttcagccc agccaggcca
     7501 ccactctgcg tgatccatga tgtgttcctc ggctgggttg accaggtggc caaggatgtg
     7561 ggctctacag gcgtggtgtt cacaacaggc ggcgcttatg gcacatccgc ctacgtgtcc
     7621 atctggaacg atctcccgca ccagaactac tccgacgacc aagagttccc gctgccaggc
     7681 ttcccagaga accataagtt ccgcaggtcc cagctccatc ggttcctcag atatgccgac
     7741 ggctccgacg attggtccaa gtatttccag ccgcagctcc gccagtccat gaagtctttt
     7801 ggctggctct gcaactccgt ggaagagatc gagacactcg gcttctccat cctccgcaac
     7861 tacaccaagc tgccgatctg gggcatcggc ccacttattg cttccccagt gcagcactcc
     7921 tcctccgaca acaattcaac aggcgccgag ttcgtgcagt ggctcagcct caaagagccg
     7981 gactccgtcc tctacatctc cttcggctcc cagaacacga tcagcccgac gcagatgatg
     8041 gaactcgctg ctggccttga gtcctccgag aagccattcc tctgggtgat cagagccccg
     8101 ttcggcttcg acatcaacga agagatgcgc ccagagtggc tgccagaggg ctttgaggaa
     8161 cgcatgaagg tgaagaaaca gggcaagctc gtgtacaagc tcggcccgca gcttgagatc
     8221 ctcaaccatg aatccatcgg cggctttctc acccactgcg gatggaacag catccttgag
     8281 tctcttcgcg agggcgttcc gatgcttgga tggccacttg ctgccgagca ggcctacaac
     8341 ctcaagtacc tcgaagatga gatgggcgtc gcggttgagc ttgctagagg cctcgaaggc
     8401 gagatctcca aagagaaggt caagcgcatc gtcgagatga tccttgagcg caacgagggc
     8461 tccaaaggct gggagatgaa gaatcgcgcc gtggaaatgg gcaaaaagct caaggacgcc
     8521 gtgaacgagg aaaaagagct gaagggctcc tccgtgaagg cgatcgacga tttcctcgac
     8581 gccgtcatgc aggccaaact tgagccaagc ctccagtgat agtgagcttg tcctgcttta
     8641 atgagatatg cgagaagcct atgatcgcat gatatttgct ttcaattctg ttgtgcacgt
     8701 tgtaaaaaac ctgagcatgt gtagctcaga tccttaccgc cggtttcggt tcattctaat
     8761 gaatatatca cccgttacta tcgtattttt atgaataata ttctccgttc aatttactga
     8821 ttgtacccta ctacttatat gtacaatatt aaaatgaaaa caatatattg tgctgaatag
     8881 gtttatagcg acatctatga tagagcgcca caataacaaa caattgcgtt ttattattac
     8941 aaatccaatt ttaaaaaaag cggcagaacc ggtcaaacct aaaagactga ttacataaat
     9001 cttattcaaa tttcaaaagt gccccagggg ctagtatcta cgacacaccg agcggcgaac
     9061 taataacgct cactgaaggg aactccggtt ccccgccggc gcgcatgggt gagattcctt
     9121 gaagttgagt attggccgtc cgctctaccg aaagttacgg gcaccattca acccggtcca
     9181 gcacggcggc cgggtaaccg acttgctgcc ccgagaatta tgcagcattt ttttggtgta
     9241 tgtgggcccc aaatgaagtg caggtcaaac cttgacagtg acgacaaatc gttgggcggg
     9301 tccagggcga attttgcgac aacatgtcga ggctcagcag gaccgct
//
