LOCUS       sequence-403395-        6090 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Backbone vector to clone toxic or unstable DNA using dual-control 
            (transcriptional control via naringenin inducible pfdeA promoter - 
            translational control via B12 riboswitch).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6090)
  AUTHORS   Vandierendonck J, Girardin Y, De Bruyn P, De Greve H, Loris R
  TITLE     A Multi-Layer-Controlled Strategy for Cloning and Expression of 
            Toxin Genes in Escherichia coli.
  JOURNAL   Toxins (Basel). 2023 Aug 18;15(8):508. doi: 10.3390/toxins15080508.
  PUBMED    37624265
REFERENCE   2  (bases 1 to 6090)
  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: "Toxins 
            (Basel)."; date: "2023-08-18"; volume: "15(8):508. doi"; pages: " 
            10.3390/toxins15080508"
COMMENT     Sequence Label: pJYP1
FEATURES             Location/Qualifiers
     source          1..6090
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(26..73)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     69..87
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(140..157)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     regulatory      1150..1159
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     terminator      complement(1468..1539)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             3252..3443
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(3437..3459)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3545..3687
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3600..3619
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3702..3719)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3873..4461)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3953..3972)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4632..5423,5424..5492))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5255..5274
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5493..5597)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5624..6079)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5761..5782)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5973..5992
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
       61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
      121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gaggctatca gcagatgcct
      181 ggcagcgccg catccatctc ctgcgcactc tccagaaaca cccgacgcag ccactggatg
      241 ccaggatcat tgctgcggta gcgatgccac tgcatcatct gccgcatctc gcccagcgac
      301 agcggactct ccttgatcac caccggccat tgcggagcca gcagctgcgc cagccgggca
      361 tgcaccgtgg cgatgcggtc cgtcccctgt accagcgcca gcgcagaagc gaagctgaag
      421 ctggtcactt ccacccggcg cgcaaagccc agcttcctgg ccatccacgc ctccaccgac
      481 gacgcattgg ccccaggcgg caccatcacc acatggcctg aggccatgta gcgttccagc
      541 gtcagctcgc cttgcgccag cgcactgtcg cgccagacca cgcagacatg ccgttcgcgg
      601 aagacctctt cggcaggatg atcgggcgtg cagaattcct gcggcaagac cagcaggtcc
      661 acctcggccc gatccagcga gcgggtcggg tcttgcacct gcggcatcag ggcaaagcgg
      721 atgtgcttgc cctcggcgtg cgcgcgcgcc agcacccggg ggatgaggac ggagagcgta
      781 aagtccgaaa ccgagatgcg aaactcgcgc gtggactcgg ccggcacgaa ggccggcagc
      841 gccgcgatgg agccatcgat acgccgcagc acatcatgca ccgcatcctt gagcacctcg
      901 gcgcgcggcg tgggctccat gcgccggccc acctggatca gcaattcatc atcgaaatac
      961 tcgcgcagcc gcgccagggc attgctcatg gccgactggc tcagatggat cttttcggcg
     1021 gcgcggctga tgctcatctc cgtgagcagt gcatccaggg cgaccagaag attgaggtcg
     1081 agcttgttga aacgcatctt tgtttcctcc gaattcgagg tggacgaatc ccaaccttct
     1141 actagagcgg ccgccatgga tatgtacaca ttatatcaca tctattccaa aatgtcaact
     1201 gaaggcgctc ctcatactgg ctgaatacat gacccgacac gttatcagaa tttatcatgt
     1261 catgattgct tacaatactc acataattgt gaaaattgta gaagccgcaa cattcaatta
     1321 tcaatcaaaa tcatgtccgc gccagtttcg ttcctcgccg ttgcactaat acagggtaat
     1381 tccctcacca cataaggaat taatgacgtc aggtggcact tttccacagc actaatacag
     1441 ggtaattccc tcaccacata aggacaggag agcgttcacc gacaaacaac agataaaacg
     1501 aaaggcccag tctttcgact gagcctttcg ttttatttga tgcctggcag ttccccaatc
     1561 agtcggaacg gcaatgctag cccatggtta ggcgctggtc tccgttgttg tgcttgttct
     1621 tgccgaccct cggatagacg acggatgggg tggtcaatgt attgatgccg tccatatcat
     1681 gaatcaaaac aatccatttg atcaatatca agctcactct taagcttcac tcatccgctg
     1741 catggcccca ccagaaaggg ctggcgcggc aagccggcgg cgcactcgca ctggatgcgc
     1801 cgctgttgag cctggccatg acaacgcgcc gatatcgccg gtcctgtgag ttaataggga
     1861 atccagtgcg aatctggagc tgacgcgcag cggtaaggaa aggtgcgatg attgcgttat
     1921 gcggacactg ccattcggtg ggaagtcatc atctcttagt atcttagata cccctccaag
     1981 cccgaagacc tgccggccaa cgtcgcatct ggttctcatc atcgcgtaat attgatgaaa
     2041 cctgcggcat ccttcttcta ttgtggatgc tttacagatc tcttcgtaaa tctggcgagt
     2101 ggggaactgc cagacatcaa ataaaacaaa aggctcagtc ggaagactgg gccttttgtt
     2161 ttatctgttg tttgtcggtg aacactctcc cgtgtaggct ggagctgctt cgaagttaac
     2221 caccatcaaa caggattttc gcctgctggg gcaaaccagc gtggaccgct tgctgcaact
     2281 ctctcagggc caggcggtga agggcaatca gctgttgccc gtctcactgg tgaaaagaaa
     2341 aaccaccctg gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     2401 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
     2461 taagttagct cactcattag gcaccgggat ctcgaccgat gcccttgaga gccttcaacc
     2521 cagtcagctc cttccggtgg gcgcggggca tgactatcgt cgccgcactt atgactgtct
     2581 tctttatcat gcaactcgta ggacaggtgc cggcagcgct ctgggtcatt ttcggcgagg
     2641 accgctttcg ctggagcgcg acgatgatcg gcctgtcgct tgcggtattc ggaatcttgc
     2701 acgccctcgc tcaagccttc gtcactggtc ccgccaccaa acgtttcggc gagaagcagg
     2761 ccattatcgc cggcatggcg gccccacggg tgcgcatgat cgtgctcctg tcgttgagga
     2821 cccggctagg ctggcggggt tgccttactg gttagcagaa tgaatcaccg atacgcgagc
     2881 gaacgtgaag cgactgctgc tgcaaaacgt ctgcgacctg agcaacaaca tgaatggtct
     2941 tcggtttccg tgtttcgtaa agtctggaaa cgcggaagtc agcgccctgc accattatgt
     3001 tccggatctg catcgcagga tgctgctggc taccctgtgg aacacctaca tctgtattaa
     3061 cgaagcgctg gcattgaccc tgagtgattt ttctctggtc ccgccgcatc cataccgcca
     3121 gttgtttacc ctcacaacgt tccagtaacc gggcatgttc atcatcagta acccgtatcg
     3181 tgagcatcct ctctcgtttc atcggtatca ttacccccat gaacagaaat cccccttaca
     3241 cggaggcatc agtgaccaaa caggaaaaaa ccgcccttaa catggcccgc tttatcagaa
     3301 gccagacatt aacgcttctg gagaaactca acgagctgga cgcggatgaa caggcagaca
     3361 tctgtgaatc gcttcacgac cacgctgatg agctttaccg cagctgcctc gcgcgtttcg
     3421 gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt
     3481 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
     3541 ggggcgcagc catgacccag tcacgtagcg atagcggagt gtatactggc ttaactatgc
     3601 ggcatcagag cagattgtac tgagagtgca ccatatatgc ggtgtgaaat accgcacaga
     3661 tgcgtaagga gaaaataccg catcaggcgc tcttccgctt cctcgctcac tgactcgctg
     3721 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     3781 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     3841 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     3901 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     3961 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     4021 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     4081 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     4141 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     4201 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     4261 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     4321 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     4381 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     4441 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     4501 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     4561 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     4621 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     4681 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     4741 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     4801 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     4861 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     4921 agtttgcgca acgttgttgc cattgctgca ggcatcgtgg tgtcacgctc gtcgtttggt
     4981 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     5041 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     5101 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     5161 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     5221 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     5281 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     5341 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     5401 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     5461 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     5521 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     5581 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgaaattgt aaacgttaat
     5641 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     5701 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     5761 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     5821 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     5881 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     5941 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     6001 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     6061 gcgccgctac agggcgcgtc ccattcgcca
//
