LOCUS       sequence-311670-        8676 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BaSP and CuCBP co-expression: araBAD promoter controls BaSP 
            expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8676)
  AUTHORS   Schwaiger KN, Voit A, Dobiasova H, Luley C, Wiltschi B, Nidetzky B
  TITLE     Plasmid Design for Tunable Two-Enzyme Co-Expression Promotes 
            Whole-Cell Production of Cellobiose.
  JOURNAL   Biotechnol J. 2020 Jul 15:e2000063. doi: 10.1002/biot.202000063.
  PUBMED    32668097
REFERENCE   2  (bases 1 to 8676)
  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: "Biotechnol 
            J. 2020 Jul 15:e2000063. doi: 10.1002/biot.202000063."
COMMENT     Sequence Label: pDUAL_araBAD
FEATURES             Location/Qualifiers
     source          1..8676
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1..285)
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     complement(59..78)
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             312..1190
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     terminator      complement(1198..1225)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     misc_feature    1293..1433
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     1348..1367
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1448..1465)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1619..2207)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1699..1718)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2244..2263
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2244..2262
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2263..2287
                     /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)."
     RBS             2302..2324
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2343..2360
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      4842..4889
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(join(4932..5723,5724..5792))
                     /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     5555..5574
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     RBS             5799..5810
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     primer_bind     6865..6884
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6904..6981)
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     terminator      complement(6988..7082)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(8610..8633)
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     RBS             8649..8660
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
ORIGIN
        1 atggagaaac agtagagagt tgcgataaaa agcgtcaggt aggatccgct aatcttatgg
       61 ataaaaatgc tatggcatag caaagtgtga cgccgtgcaa ataatcaatg tggacttttc
      121 tgccgtgatt atagacactt ttgttacgcg tttttgtcat ggctttggtc ccgctttgtt
      181 acagaatgct tttaataagc ggggttaccg gtttggttag cgagaagagc cagtaaaaga
      241 cgcagtgacg gcaatgtctg atgcaatatg gacaattggt ttcttctctg aatggcggga
      301 gtatgaaaag tatggctgaa gcgcaaaatg atcccctgct gccgggatac tcgtttaatg
      361 cccatctggt ggcgggttta acgccgattg aggccaacgg ttatctcgat ttttttatcg
      421 accgaccgct gggaatgaaa ggttatattc tcaatctcac cattcgcggt cagggggtgg
      481 tgaaaaatca gggacgagaa tttgtttgcc gaccgggtga tattttgctg ttcccgccag
      541 gagagattca tcactacggt cgtcatccgg aggctcgcga atggtatcac cagtgggttt
      601 actttcgtcc gcgcgcctac tggcatgaat ggcttaactg gccgtcaata tttgccaata
      661 cggggttctt tcgcccggat gaagcgcacc agccgcattt cagcgacctg tttgggcaaa
      721 tcattaacgc cgggcaaggg gaagggcgct attcggagct gctggcgata aatctgcttg
      781 agcaattgtt actgcggcgc atggaagcga ttaacgagtc gctccatcca ccgatggata
      841 atcgggtacg cgaggcttgt cagtacatca gcgatcacct ggcagacagc aattttgata
      901 tcgccagcgt cgcacagcat gtttgcttgt cgccgtcgcg tctgtcacat cttttccgcc
      961 agcagttagg gattagcgtc ttaagctggc gcgaggacca acgtatcagc caggcgaagc
     1021 tgcttttgag caccacccgg atgcctatcg ccaccgtcgg tcgcaatgtt ggttttgacg
     1081 atcaactcta tttctcgcgg gtatttaaaa aatgcaccgg ggccagcccg agcgagttcc
     1141 gtgccggttg tgaagaaaaa gtgaatgatg tagccgtcaa gttgtcataa ggagctcaaa
     1201 aggccatccg tcaggatggc cttcttccgc gcacatttcc ccgaaaagtg ccacctgcat
     1261 cgatttagcg gccgcgttgg cgggtgtcgg ggcgcagcca tgacccagtc acgtagcgat
     1321 agcggagtgt atactggctt aactatgcgg catcagagca gattgtactg agagtgcacc
     1381 atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgctctt
     1441 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     1501 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     1561 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     1621 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     1681 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     1741 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     1801 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     1861 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     1921 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     1981 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     2041 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     2101 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     2161 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     2221 tctactagtc gatcccgcga aattaatacg actcactata ggggaattgt gagcggataa
     2281 caattcccct ctagaaataa ttttgtttaa ctttaagaag gagatatcat atgggcagca
     2341 gccatcacca tcaccatcac gggtcgaccc gctatggtca ttttgacgat gaagcacgcg
     2401 agtacgttat caccacaccg catactccgt acccttggat caactatctg ggttcggaac
     2461 agttcttcag tcttttgtcg caccaggctg gcggctattc gttttatcgc gatgccaaga
     2521 tgcgtagatt gacaagatac agatacaaca acataccagc agacgcggga ggccggtatc
     2581 tttatgtaaa cgacggtggg gacgtttgga cgccatcgtg gcttcctgtt aaagcagact
     2641 tagatcattt tgaagccaga catggtcttg gctacagtac cattacaggt gaaagaaatg
     2701 gggtacgggt tgagaccttg tttttcgtcc cagtcggtga gaacgcggag gtccagaaag
     2761 ttacggtcac taatacctct gactcgtata aatccctgac gttgttttct tttgtcgaat
     2821 tttgtctttg gaacgcgcaa gacgatcaaa ctaactacca gcgtaatctg agcataggag
     2881 aggttgaagt ggaacaagaa tctccgcatg gcagtgcaat ttatcaccgt acagagtacc
     2941 gggagagacg cgatcactac gcggtattcg ccgttaatac gcaggccgag ggtttcgata
     3001 cagatcgcga cacatttgtg ggagcctata actcgcttgg tgaagccgca gttcccctta
     3061 agggagaaag tgcaaatagc gttgcaagtg gatggtatcc tattggctct catagcgtgg
     3121 cagtgtcttt agcgcctgga gaatccagag aattggttta tgtgcttggg tacgtggaaa
     3181 atcctgatga ggagaagtgg gcagacgacg caaaacaggt cgtgaataaa gaaagagctc
     3241 acgctttatt atcaagattt gctacgtccg aacagactga cgcagctttc gccgctttga
     3301 aggattattg gacggatttg ttatcgacgt attccgttag ttccaatgac gaaaaattgg
     3361 accggatggt taatatatgg aatcagtacc aatgcatggt aacctttaac atgagtcgct
     3421 cggcgagctt tttcgagacc ggtattggga gagggatggg gtttcgtgat agcaaccaag
     3481 accttctggg ctttgttcac ttaatcccag agcgggcgcg tgagcgcatc attgacatag
     3541 cgtcaaccca gttcgcggac ggttccgctt atcaccaata ccaaccttta acgaagcgcg
     3601 gtaacaacga cattggctct gggtttaacg acgatcctct ttggttaatc gcgggaactg
     3661 ccgcctacat caaagaaact ggtgacttct ccatattaga tgagcccgta ccttttgata
     3721 acgaaccggg gtcagaagtt ccgttatttg agcacctgac ccgtagtttt gagtttaccg
     3781 taacccatcg gggtccccac ggtttgcctc ttattggccg cgcagattgg aacgattgtt
     3841 tgaatcttaa ttgcttcagt acaaccccgg gcgagtcatt tcagacaacg gagaaccaag
     3901 ccggtggcgt agcggagtca acgttcatag ccgcgcaatt cgttttatat ggggagcagt
     3961 atgccgagtt ggcagcgcgg cgcgggttgg ccgacgtagc cgatagagca agagggcatg
     4021 tggctgaaat gagagacgca cttctgaccg acgggtggga cggcagctgg ttcttgagag
     4081 cttatgacta ttatggaaac cctatcggca ccgacgcaca tgacgaaggg aagatttgga
     4141 tcgaaccgca agggttcgcg gtgatggctg gagtcggtgt aggcgaaggg cctcaagata
     4201 cggacgctcc cgctattaag gcgcttgatt ctgtgaacga aatgttggca accgatcacg
     4261 gtatggtcct tcaataccca gcatacacta cataccaagt acacatgggt gaagtatcca
     4321 cttatccacc cgggtataaa gagaatggag gtatattttg ccacaataat ccttgggtta
     4381 tcatagcgga aactgtcgtc ggaagaggcg gacgggcatt tgattactac aagagaatta
     4441 ctcccgcgta tcgcgaggac ataagcgacg ttcaccgtct tgaaccctat gtctatgcac
     4501 aaatgatagc agggaaggag gcggtccgtc acggtgaagc aaagaacagc tggcttaccg
     4561 ggactgcggc ttggaacttt gtgaccgtgt cacagtacct tcttggggtt cgcccagagt
     4621 acgacggctt ggtggtagac ccgcagatcg gacctgacgt acctagcttt acggttacgc
     4681 gtgtcgcccg tggagccact tacgagataa cagtcactaa cagtgggacc gacggatcac
     4741 gcggtcgcct tgtcgttgat ggaacacctg tagaaggcaa cttggttcct tatgcccccg
     4801 cagggtcaac agttcgtgtc gatgtgacat tgtagtgtac actagcataa ccccttgggg
     4861 cctctaaacg ggtcttgagg ggttttttgc tcgagtaaaa aaaatcctta gctttcgcta
     4921 aggatctgca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     4981 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     5041 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     5101 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     5161 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     5221 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     5281 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     5341 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     5401 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     5461 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     5521 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     5581 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     5641 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     5701 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     5761 ataagggcga cacggaaatg ttgaatactc atgcttaatt tctcctcttt aatagatctt
     5821 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     5881 gcgcggggag aggcggtttg cgtattgggc gccagggtgg tttttctttt caccagtgag
     5941 acgggcaaca gctgattgcc cttcaccgcc tggccctgag agagttgcag caagcggtcc
     6001 acgctggttt gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa
     6061 catgagctgt cctcggtatc gtcgtatccc actaccgaga tatccgcacc aacgcgcagc
     6121 ccggactcgg taatggcgcg cattgcgccc agcgccatct gatcgttggc aaccagcatc
     6181 gcagtgggaa cgatgccctc attcagcatt tgcatggttt gttgaaaacc ggacatggca
     6241 ctccagtcgc cttcccgttc cgctatcggc tgaatttgat tgcgagtgag atatttatgc
     6301 cagccagcca gacgcagacg cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt
     6361 tgctggtgac ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc ctcatgggag
     6421 aaaataatac tgttgatggg atgtctggtc agagacatca agaaataacg ccggaacatt
     6481 agtgcaggca gcttccacag caatggcatc ctggtcatcc agcggatagt taatgatcag
     6541 cccactgacg cgttgcgcga gaagattgtg caccgccgct ttacaggctt cgacgccgct
     6601 tcgttctacc atcgacacca ccacgctggc acccagttga tcggcgcgag atttaatcgc
     6661 cgcgacaatt tgcgacggcg cgtgcagggc cagactggag gtggcaacgc caatcagcaa
     6721 cgactgtttg cccgccagtt gttgtgccac gcggttggga atgtaattca gctccgccat
     6781 cgccgcttcc actttttccc gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg
     6841 ggaaacggtc tgataagaga caccggcata ctctgcgaca tcgtataacg ttactggttt
     6901 cacattcacc accctgaatt gactctcttc cgggcgctat catgccatac cgcgaaaggt
     6961 tttgcaccat tcgatggtgt cgaattcatt ctcaccaata aaaaacgccc ggcggcaacc
     7021 gagcgttctg aacaaatcca gatggagttc tgaggtcatt actggatcta tcaacaggag
     7081 tcaagctttt acgcaacaac cggcggatta gcgatcaggt cgtcggagcg atggtcacca
     7141 gcactgtctt cccattccag cattgcgacc ggcgtggtat tatcaacacc caggccgcgt
     7201 ttcggttcga acgtcagggt ggcctgcgac gtttcaccac gccaggtaaa ggaaatactc
     7261 gtatcgtcat ccgtggtata tgagaaggtg ccgtcaaaag catccagttc attgcgaaat
     7321 ttcgccagag cgttcagtgc tttaaccacc ggacgtttca gattttcatc gatttcggcc
     7381 gtgctgtaat aatggcggtt aatgtcacga ccattgttgg ttttgcgcag cagttccata
     7441 tcatttttac ctgccagggc gcccacgtaa tagacttgcg gaacacccgg caggaaaaac
     7501 tgcaccgcac gcgcagcgat gtagtgctga tcgttacagc ccagagcgga gtagtaggta
     7561 ctattaactt ggtacagatc caggttcgat gccgccgcac cggtagctgc ctgtgattcg
     7621 ccgtgggtgt tcgcatgaat cgtattcacc aggttatcca catcttcgtc cggcaccaga
     7681 cctttcagcg agcggtccag ctgatctgaa ccgatatcaa taacgccgat gccgtcatgc
     7741 gtatccagga cggtaacagc attgttcgga cgaatatccg tccaatgggc caccggttcg
     7801 acgtggccgg tactcagggc atgcagcagc agcggcggca gggcaaagtc ataaacgcga
     7861 tccactttag acgcgatttc cacctgtttt ttgtaatagc tgtggacttc gatcagaatt
     7921 tccagaccgc gtttcacgcc ttcttcgcgc agacgggaaa tcagtttgaa cgttttcggg
     7981 gtcataaagc aactggtgcc tgcttctttg gcaccgtagc caaccgcatc caggcggata
     8041 taagacacat ggctggccgc catttggtcg aaaatggaca tcagatattc ccaaccttta
     8101 tcactgtccg tatcgatgtc cacttgctgc ggggtaaaag acacccagac cagacgcgtt
     8161 ttgcctgcga atttgtagtg ggtaaacggc agacccggac gcggacgata aatgcctgcc
     8221 aggtcttctt ccgtggcacc gttcgggaac acgctgctca tggtcagaaa catcgggtaa
     8281 tattctgatt cttcgccttt ggccagaaca tcttggaact gtttcgattc ccatgacatg
     8341 tgattaacga ttgcatccac cataatgttg tgggttttag acagttcggc gacgtcatcc
     8401 cagctaccca gacgttcatc aactttggta tggtcgatcg gatcgaaacc tgcgtcggcg
     8461 ccatcaaacg gggtgaaaaa cggcagaatg tggacaccgt cataaacgcc atcaaagcgg
     8521 gtacgcagga tgtccgtcat gcttttaatg gtgccatcac ccagacgatc cgcataggta
     8581 atcagttgga ctttattttt catggtaccc ttttcgaact gcgggtggct ccagcttgcc
     8641 atggagtctt tctcctcttt atccaaaaaa acgggt
//
