LOCUS       sequence-454070-        9405 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 1, Position 3 containing CBS4-miniDFR::RUBY::OCS..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9405)
  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 9405)
  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: pCHW91013
FEATURES             Location/Qualifiers
     source          1..9405
                     /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             4727..8677
                     /label=RUBY
     CDS             6227..6283
                     /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             7118..7174
                     /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      8691..9398
                     /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 gtcctaatac ctctacgtgc aagcagtata aaaacaaaat acataaacat
     4621 aactaaaatc tcatatcgtt gtaattgtca tatactagtg gtaattaagt ccagttacac
     4681 ttaaggttaa atttgctgac tcttttttac ctttcatttc tgaaaaatgg atcatgcgac
     4741 cctcgccatg atcctcgcga tctggttcat cagcttccac ttcatcaagc tgctgttctc
     4801 ccagcagacc accaagctgc ttccgccagg accaaagccg cttccgatca tcggcaacat
     4861 ccttgaggtg ggcaagaagc cgcatcggtc cttcgccaac ctcgccaaga ttcacggccc
     4921 actcatttcc ctcagactcg gctctgtgac caccatcgtt gtgtcctctg ccgacgtggc
     4981 caaagagatg ttcctcaaga aggatcaccc gctctccaac cgcacgatcc cgaatagtgt
     5041 tacagccggc gaccaccaca agctcaccat gtcttggctc ccggtgtctc cgaagtggcg
     5101 caacttccgc aagattaccg ccgtgcatct gctctcccca cagagactcg atgcctgcca
     5161 gacattcagg cacgccaagg tgcagcagct ctacgagtac gttcaagagt gcgcccagaa
     5221 aggccaggcc gtggatattg gcaaggccgc ctttacgacc agcctcaacc tcctcagcaa
     5281 gctgttcttc agcgtcgagc tggcgcacca caagtcccat accagccaag agttcaaaga
     5341 gctgatctgg aacatcatgg aagatatagg caagccgaac tacgccgact acttcccgat
     5401 tctcggctgc gttgacccat ctggcattag aagaaggctc gcctgctcct tcgacaagct
     5461 gatcgccgtg ttccagggca tcatctgcga gagactcgcc ccagattcct ccaccacaac
     5521 taccaccacc accgacgacg tgctcgatgt gctcctccag ctgttcaagc agaacgagct
     5581 gacgatgggc gagatcaacc acctcctcgt ggacatcttc gacgccggca ccgataccac
     5641 atcctccaca ttcgagtggg tgatgaccga gctgatccgc aatccagaga tgatggaaaa
     5701 ggcccaagag gaaatcaagc aggtcctcgg caaggacaag cagatccaag agtccgacat
     5761 catcaacctg ccgtacctcc aggcgatcat caaagagaca ctccgcctcc atccgccgac
     5821 cgtgttcttg ctcccaagaa aggccgacac cgatgtcgag ctgtacggct acatcgtgcc
     5881 gaaggatgcc cagatcctcg tgaacctctg ggccattggc agggacccaa acgcctggca
     5941 gaacgccgat attttcagcc cagagcgctt catcggctgc gagatcgatg ttaagggccg
     6001 cgatttcggc ctccttccat ttggcgctgg ccgcagaatt tgcccaggca tgaatctcgc
     6061 catcaggatg ctcaccctca tgctcgccac actcctccag ttcttcaact ggaagctcga
     6121 aggcgacatc tccccgaagg acctcgacat ggacgagaag ttcggcattg cgctccaaaa
     6181 gaccaagccg ctcaagctca tcccgattcc gcgctacggt agcggagcta ccaattttag
     6241 cctccttaag caggcaggtg atgtagaaga gaaccccggg cctatgaaga tgatgaacgg
     6301 cgaggacgcc aacgaccaga tgatcaaaga gtccttcttc atcacccacg gcaacccgat
     6361 cctcaccgtc gaggatacac atccgctcag gccgttcttc gagacatggc gcgagaagat
     6421 tttctccaag aagccgaagg ccatcctcat catctccggc cactgggaga cagtgaagcc
     6481 aaccgtgaac gccgtgcaca tcaacgacac catccacgac ttcgacgact acccagccgc
     6541 catgtaccag ttcaagtacc cagctccagg cgagccagag cttgcgagaa aggtggaaga
     6601 gatcctcaag aagtccgggt tcgagacagc cgagacagac caaaagaggg gccttgatca
     6661 cggcgcctgg gttccactca tgctcatgta tccagaggcg gacatcccgg tgtgccagct
     6721 ctcagttcag ccacatctcg acggcaccta ccactacaat ctcggcagag ccctcgcgcc
     6781 gctcaagaat gatggcgtgc tcattattgg ctccggcagc gccacacatc cactcgatga
     6841 gacaccgcac tacttcgatg gtgttgcccc ttgggccgct gccttcgatt cttggcttag
     6901 gaaggccctc atcaacggcc gcttcgagga agtgaacatc tacgagagca aggccccgaa
     6961 ctggaagctc gcccatccat ttccagagca cttctacccg ctccacgttg tgctcggcgc
     7021 tgctggtgaa aagtggaagg ccgagctgat ccactcctcc tgggatcatg gcacactttg
     7081 ccacggctcc tacaagttca cctccgccgg atccggagca accaacttta gcctgctcaa
     7141 gcaagcagga gatgttgagg aaaatcctgg ccccatgacc gccatcaaga tgaacaccaa
     7201 cggcgagggc gagacacagc acatcctcat gatcccgttc atggcgcagg gccacctcag
     7261 gccatttctc gaactcgcca tgttcctcta caagcgctcc cacgtgatca tcaccctgct
     7321 cacaactccg ctcaatgccg gcttcctcag gcacctcctt caccaccatt cctactcctc
     7381 cagcggcatc aggatcgtcg agctgccatt caactccacc aaccacggac tcccaccggg
     7441 catcgagaac accgataagc tcacactccc gctcgtggtg tccctcttcc attccaccat
     7501 cagcctcgat ccgcacctcc gcgattacat ctccaggcat ttcagcccag ccaggccacc
     7561 actctgcgtg atccatgatg tgttcctcgg ctgggttgac caggtggcca aggatgtggg
     7621 ctctacaggc gtggtgttca caacaggcgg cgcttatggc acatccgcct acgtgtccat
     7681 ctggaacgat ctcccgcacc agaactactc cgacgaccaa gagttcccgc tgccaggctt
     7741 cccagagaac cataagttcc gcaggtccca gctccatcgg ttcctcagat atgccgacgg
     7801 ctccgacgat tggtccaagt atttccagcc gcagctccgc cagtccatga agtcttttgg
     7861 ctggctctgc aactccgtgg aagagatcga gacactcggc ttctccatcc tccgcaacta
     7921 caccaagctg ccgatctggg gcatcggccc acttattgct tccccagtgc agcactcctc
     7981 ctccgacaac aattcaacag gcgccgagtt cgtgcagtgg ctcagcctca aagagccgga
     8041 ctccgtcctc tacatctcct tcggctccca gaacacgatc agcccgacgc agatgatgga
     8101 actcgctgct ggccttgagt cctccgagaa gccattcctc tgggtgatca gagccccgtt
     8161 cggcttcgac atcaacgaag agatgcgccc agagtggctg ccagagggct ttgaggaacg
     8221 catgaaggtg aagaaacagg gcaagctcgt gtacaagctc ggcccgcagc ttgagatcct
     8281 caaccatgaa tccatcggcg gctttctcac ccactgcgga tggaacagca tccttgagtc
     8341 tcttcgcgag ggcgttccga tgcttggatg gccacttgct gccgagcagg cctacaacct
     8401 caagtacctc gaagatgaga tgggcgtcgc ggttgagctt gctagaggcc tcgaaggcga
     8461 gatctccaaa gagaaggtca agcgcatcgt cgagatgatc cttgagcgca acgagggctc
     8521 caaaggctgg gagatgaaga atcgcgccgt ggaaatgggc aaaaagctca aggacgccgt
     8581 gaacgaggaa aaagagctga agggctcctc cgtgaaggcg atcgacgatt tcctcgacgc
     8641 cgtcatgcag gccaaacttg agccaagcct ccagtgatag tgagcttgtc ctgctttaat
     8701 gagatatgcg agaagcctat gatcgcatga tatttgcttt caattctgtt gtgcacgttg
     8761 taaaaaacct gagcatgtgt agctcagatc cttaccgccg gtttcggttc attctaatga
     8821 atatatcacc cgttactatc gtatttttat gaataatatt ctccgttcaa tttactgatt
     8881 gtaccctact acttatatgt acaatattaa aatgaaaaca atatattgtg ctgaataggt
     8941 ttatagcgac atctatgata gagcgccaca ataacaaaca attgcgtttt attattacaa
     9001 atccaatttt aaaaaaagcg gcagaaccgg tcaaacctaa aagactgatt acataaatct
     9061 tattcaaatt tcaaaagtgc cccaggggct agtatctacg acacaccgag cggcgaacta
     9121 ataacgctca ctgaagggaa ctccggttcc ccgccggcgc gcatgggtga gattccttga
     9181 agttgagtat tggccgtccg ctctaccgaa agttacgggc accattcaac ccggtccagc
     9241 acggcggccg ggtaaccgac ttgctgcccc gagaattatg cagcattttt ttggtgtatg
     9301 tgggccccaa atgaagtgca ggtcaaacct tgacagtgac gacaaatcgt tgggcgggtc
     9361 cagggcgaat tttgcgacaa catgtcgagg ctcagcagga ccgct
//
