LOCUS       sequence-310563-        8676 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  see pDUAL_araBAD: exchanged origin of replication (pBR322) and 
            introduced bom site.
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_2_araBAD
FEATURES             Location/Qualifiers
     source          1..8676
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             10..21
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     promoter        complement(38..322)
                     /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(96..115)
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             349..1227
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     terminator      complement(1235..1262)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     misc_feature    1330..1470
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     1385..1404
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1485..1502)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1656..2244)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1736..1755)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2281..2300
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2281..2299
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2300..2324
                     /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             2339..2361
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2380..2397
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      4879..4926
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(join(4969..5760,5761..5829))
                     /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     5592..5611
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     RBS             5836..5847
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     primer_bind     6902..6921
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6941..7018)
                     /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(7025..7119)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(8647..8670)
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
ORIGIN
        1 catggagtct ttctcctctt tatccaaaaa aacgggtatg gagaaacagt agagagttgc
       61 gataaaaagc gtcaggtagg atccgctaat cttatggata aaaatgctat ggcatagcaa
      121 agtgtgacgc cgtgcaaata atcaatgtgg acttttctgc cgtgattata gacacttttg
      181 ttacgcgttt ttgtcatggc tttggtcccg ctttgttaca gaatgctttt aataagcggg
      241 gttaccggtt tggttagcga gaagagccag taaaagacgc agtgacggca atgtctgatg
      301 caatatggac aattggtttc ttctctgaat ggcgggagta tgaaaagtat ggctgaagcg
      361 caaaatgatc ccctgctgcc gggatactcg tttaatgccc atctggtggc gggtttaacg
      421 ccgattgagg ccaacggtta tctcgatttt tttatcgacc gaccgctggg aatgaaaggt
      481 tatattctca atctcaccat tcgcggtcag ggggtggtga aaaatcaggg acgagaattt
      541 gtttgccgac cgggtgatat tttgctgttc ccgccaggag agattcatca ctacggtcgt
      601 catccggagg ctcgcgaatg gtatcaccag tgggtttact ttcgtccgcg cgcctactgg
      661 catgaatggc ttaactggcc gtcaatattt gccaatacgg ggttctttcg cccggatgaa
      721 gcgcaccagc cgcatttcag cgacctgttt gggcaaatca ttaacgccgg gcaaggggaa
      781 gggcgctatt cggagctgct ggcgataaat ctgcttgagc aattgttact gcggcgcatg
      841 gaagcgatta acgagtcgct ccatccaccg atggataatc gggtacgcga ggcttgtcag
      901 tacatcagcg atcacctggc agacagcaat tttgatatcg ccagcgtcgc acagcatgtt
      961 tgcttgtcgc cgtcgcgtct gtcacatctt ttccgccagc agttagggat tagcgtctta
     1021 agctggcgcg aggaccaacg tatcagccag gcgaagctgc ttttgagcac cacccggatg
     1081 cctatcgcca ccgtcggtcg caatgttggt tttgacgatc aactctattt ctcgcgggta
     1141 tttaaaaaat gcaccggggc cagcccgagc gagttccgtg ccggttgtga agaaaaagtg
     1201 aatgatgtag ccgtcaagtt gtcataagga gctcaaaagg ccatccgtca ggatggcctt
     1261 cttccgcgca catttccccg aaaagtgcca cctgcatcga tttagcggcc gcgttggcgg
     1321 gtgtcggggc gcagccatga cccagtcacg tagcgatagc ggagtgtata ctggcttaac
     1381 tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac
     1441 agatgcgtaa ggagaaaata ccgcatcagg cgctcttccg cttcctcgct cactgactcg
     1501 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     1561 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     1621 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     1681 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     1741 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     1801 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     1861 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     1921 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     1981 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     2041 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     2101 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     2161 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     2221 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct actagtcgat cccgcgaaat
     2281 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
     2341 tgtttaactt taagaaggag atatcatatg ggcagcagcc atcaccatca ccatcacggg
     2401 tcgacccgct atggtcattt tgacgatgaa gcacgcgagt acgttatcac cacaccgcat
     2461 actccgtacc cttggatcaa ctatctgggt tcggaacagt tcttcagtct tttgtcgcac
     2521 caggctggcg gctattcgtt ttatcgcgat gccaagatgc gtagattgac aagatacaga
     2581 tacaacaaca taccagcaga cgcgggaggc cggtatcttt atgtaaacga cggtggggac
     2641 gtttggacgc catcgtggct tcctgttaaa gcagacttag atcattttga agccagacat
     2701 ggtcttggct acagtaccat tacaggtgaa agaaatgggg tacgggttga gaccttgttt
     2761 ttcgtcccag tcggtgagaa cgcggaggtc cagaaagtta cggtcactaa tacctctgac
     2821 tcgtataaat ccctgacgtt gttttctttt gtcgaatttt gtctttggaa cgcgcaagac
     2881 gatcaaacta actaccagcg taatctgagc ataggagagg ttgaagtgga acaagaatct
     2941 ccgcatggca gtgcaattta tcaccgtaca gagtaccggg agagacgcga tcactacgcg
     3001 gtattcgccg ttaatacgca ggccgagggt ttcgatacag atcgcgacac atttgtggga
     3061 gcctataact cgcttggtga agccgcagtt ccccttaagg gagaaagtgc aaatagcgtt
     3121 gcaagtggat ggtatcctat tggctctcat agcgtggcag tgtctttagc gcctggagaa
     3181 tccagagaat tggtttatgt gcttgggtac gtggaaaatc ctgatgagga gaagtgggca
     3241 gacgacgcaa aacaggtcgt gaataaagaa agagctcacg ctttattatc aagatttgct
     3301 acgtccgaac agactgacgc agctttcgcc gctttgaagg attattggac ggatttgtta
     3361 tcgacgtatt ccgttagttc caatgacgaa aaattggacc ggatggttaa tatatggaat
     3421 cagtaccaat gcatggtaac ctttaacatg agtcgctcgg cgagcttttt cgagaccggt
     3481 attgggagag ggatggggtt tcgtgatagc aaccaagacc ttctgggctt tgttcactta
     3541 atcccagagc gggcgcgtga gcgcatcatt gacatagcgt caacccagtt cgcggacggt
     3601 tccgcttatc accaatacca acctttaacg aagcgcggta acaacgacat tggctctggg
     3661 tttaacgacg atcctctttg gttaatcgcg ggaactgccg cctacatcaa agaaactggt
     3721 gacttctcca tattagatga gcccgtacct tttgataacg aaccggggtc agaagttccg
     3781 ttatttgagc acctgacccg tagttttgag tttaccgtaa cccatcgggg tccccacggt
     3841 ttgcctctta ttggccgcgc agattggaac gattgtttga atcttaattg cttcagtaca
     3901 accccgggcg agtcatttca gacaacggag aaccaagccg gtggcgtagc ggagtcaacg
     3961 ttcatagccg cgcaattcgt tttatatggg gagcagtatg ccgagttggc agcgcggcgc
     4021 gggttggccg acgtagccga tagagcaaga gggcatgtgg ctgaaatgag agacgcactt
     4081 ctgaccgacg ggtgggacgg cagctggttc ttgagagctt atgactatta tggaaaccct
     4141 atcggcaccg acgcacatga cgaagggaag atttggatcg aaccgcaagg gttcgcggtg
     4201 atggctggag tcggtgtagg cgaagggcct caagatacgg acgctcccgc tattaaggcg
     4261 cttgattctg tgaacgaaat gttggcaacc gatcacggta tggtccttca atacccagca
     4321 tacactacat accaagtaca catgggtgaa gtatccactt atccacccgg gtataaagag
     4381 aatggaggta tattttgcca caataatcct tgggttatca tagcggaaac tgtcgtcgga
     4441 agaggcggac gggcatttga ttactacaag agaattactc ccgcgtatcg cgaggacata
     4501 agcgacgttc accgtcttga accctatgtc tatgcacaaa tgatagcagg gaaggaggcg
     4561 gtccgtcacg gtgaagcaaa gaacagctgg cttaccggga ctgcggcttg gaactttgtg
     4621 accgtgtcac agtaccttct tggggttcgc ccagagtacg acggcttggt ggtagacccg
     4681 cagatcggac ctgacgtacc tagctttacg gttacgcgtg tcgcccgtgg agccacttac
     4741 gagataacag tcactaacag tgggaccgac ggatcacgcg gtcgccttgt cgttgatgga
     4801 acacctgtag aaggcaactt ggttccttat gcccccgcag ggtcaacagt tcgtgtcgat
     4861 gtgacattgt agtgtacact agcataaccc cttggggcct ctaaacgggt cttgaggggt
     4921 tttttgctcg agtaaaaaaa atccttagct ttcgctaagg atctgcagtt accaatgctt
     4981 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     5041 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     5101 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     5161 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     5221 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     5281 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     5341 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     5401 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     5461 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     5521 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     5581 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     5641 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     5701 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     5761 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     5821 aatactcatg cttaatttct cctctttaat agatcttcac tgcccgcttt ccagtcggga
     5881 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     5941 attgggcgcc agggtggttt ttcttttcac cagtgagacg ggcaacagct gattgccctt
     6001 caccgcctgg ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg
     6061 aaaatcctgt ttgatggtgg ttaacggcgg gatataacat gagctgtcct cggtatcgtc
     6121 gtatcccact accgagatat ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat
     6181 tgcgcccagc gccatctgat cgttggcaac cagcatcgca gtgggaacga tgccctcatt
     6241 cagcatttgc atggtttgtt gaaaaccgga catggcactc cagtcgcctt cccgttccgc
     6301 tatcggctga atttgattgc gagtgagata tttatgccag ccagccagac gcagacgcgc
     6361 cgagacagaa cttaatgggc ccgctaacag cgcgatttgc tggtgaccca atgcgaccag
     6421 atgctccacg cccagtcgcg taccgtcctc atgggagaaa ataatactgt tgatgggatg
     6481 tctggtcaga gacatcaaga aataacgccg gaacattagt gcaggcagct tccacagcaa
     6541 tggcatcctg gtcatccagc ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa
     6601 gattgtgcac cgccgcttta caggcttcga cgccgcttcg ttctaccatc gacaccacca
     6661 cgctggcacc cagttgatcg gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt
     6721 gcagggccag actggaggtg gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt
     6781 gtgccacgcg gttgggaatg taattcagct ccgccatcgc cgcttccact ttttcccgcg
     6841 ttttcgcaga aacgtggctg gcctggttca ccacgcggga aacggtctga taagagacac
     6901 cggcatactc tgcgacatcg tataacgtta ctggtttcac attcaccacc ctgaattgac
     6961 tctcttccgg gcgctatcat gccataccgc gaaaggtttt gcaccattcg atggtgtcga
     7021 attcattctc accaataaaa aacgcccggc ggcaaccgag cgttctgaac aaatccagat
     7081 ggagttctga ggtcattact ggatctatca acaggagtca agcttttacg caacaaccgg
     7141 cggattagcg atcaggtcgt cggagcgatg gtcaccagca ctgtcttccc attccagcat
     7201 tgcgaccggc gtggtattat caacacccag gccgcgtttc ggttcgaacg tcagggtggc
     7261 ctgcgacgtt tcaccacgcc aggtaaagga aatactcgta tcgtcatccg tggtatatga
     7321 gaaggtgccg tcaaaagcat ccagttcatt gcgaaatttc gccagagcgt tcagtgcttt
     7381 aaccaccgga cgtttcagat tttcatcgat ttcggccgtg ctgtaataat ggcggttaat
     7441 gtcacgacca ttgttggttt tgcgcagcag ttccatatca tttttacctg ccagggcgcc
     7501 cacgtaatag acttgcggaa cacccggcag gaaaaactgc accgcacgcg cagcgatgta
     7561 gtgctgatcg ttacagccca gagcggagta gtaggtacta ttaacttggt acagatccag
     7621 gttcgatgcc gccgcaccgg tagctgcctg tgattcgccg tgggtgttcg catgaatcgt
     7681 attcaccagg ttatccacat cttcgtccgg caccagacct ttcagcgagc ggtccagctg
     7741 atctgaaccg atatcaataa cgccgatgcc gtcatgcgta tccaggacgg taacagcatt
     7801 gttcggacga atatccgtcc aatgggccac cggttcgacg tggccggtac tcagggcatg
     7861 cagcagcagc ggcggcaggg caaagtcata aacgcgatcc actttagacg cgatttccac
     7921 ctgttttttg taatagctgt ggacttcgat cagaatttcc agaccgcgtt tcacgccttc
     7981 ttcgcgcaga cgggaaatca gtttgaacgt tttcggggtc ataaagcaac tggtgcctgc
     8041 ttctttggca ccgtagccaa ccgcatccag gcggatataa gacacatggc tggccgccat
     8101 ttggtcgaaa atggacatca gatattccca acctttatca ctgtccgtat cgatgtccac
     8161 ttgctgcggg gtaaaagaca cccagaccag acgcgttttg cctgcgaatt tgtagtgggt
     8221 aaacggcaga cccggacgcg gacgataaat gcctgccagg tcttcttccg tggcaccgtt
     8281 cgggaacacg ctgctcatgg tcagaaacat cgggtaatat tctgattctt cgcctttggc
     8341 cagaacatct tggaactgtt tcgattccca tgacatgtga ttaacgattg catccaccat
     8401 aatgttgtgg gttttagaca gttcggcgac gtcatcccag ctacccagac gttcatcaac
     8461 tttggtatgg tcgatcggat cgaaacctgc gtcggcgcca tcaaacgggg tgaaaaacgg
     8521 cagaatgtgg acaccgtcat aaacgccatc aaagcgggta cgcaggatgt ccgtcatgct
     8581 tttaatggtg ccatcaccca gacgatccgc ataggtaatc agttggactt tatttttcat
     8641 ggtacccttt tcgaactgcg ggtggctcca gcttgc
//
