LOCUS       sequence-300116-        8409 bp    DNA     circular SYN 15-APR-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 8409)
  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 8409)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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..8409
                     /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"
     rep_origin      1348..1936
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1837..1856
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2015..2034
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2015..2033
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2034..2058
                     /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             2073..2095
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2114..2131
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      4613..4660
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(join(4703..5494,5495..5563))
                     /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     5326..5345
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     RBS             5570..5581
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             complement(5591..6673)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     6635..6654
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6674..6751)
                     /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(6758..6852)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(8380..8403)
                     /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 gcagaagatc
     1321 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     1381 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     1441 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     1501 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     1561 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     1621 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     1681 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     1741 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     1801 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     1861 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     1921 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     1981 gccttttgct cacaactagt cgatcccgcg aaattaatac gactcactat aggggaattg
     2041 tgagcggata acaattcccc tctagaaata attttgttta actttaagaa ggagatatca
     2101 tatgggcagc agccatcacc atcaccatca cgggtcgacc cgctatggtc attttgacga
     2161 tgaagcacgc gagtacgtta tcaccacacc gcatactccg tacccttgga tcaactatct
     2221 gggttcggaa cagttcttca gtcttttgtc gcaccaggct ggcggctatt cgttttatcg
     2281 cgatgccaag atgcgtagat tgacaagata cagatacaac aacataccag cagacgcggg
     2341 aggccggtat ctttatgtaa acgacggtgg ggacgtttgg acgccatcgt ggcttcctgt
     2401 taaagcagac ttagatcatt ttgaagccag acatggtctt ggctacagta ccattacagg
     2461 tgaaagaaat ggggtacggg ttgagacctt gtttttcgtc ccagtcggtg agaacgcgga
     2521 ggtccagaaa gttacggtca ctaatacctc tgactcgtat aaatccctga cgttgttttc
     2581 ttttgtcgaa ttttgtcttt ggaacgcgca agacgatcaa actaactacc agcgtaatct
     2641 gagcatagga gaggttgaag tggaacaaga atctccgcat ggcagtgcaa tttatcaccg
     2701 tacagagtac cgggagagac gcgatcacta cgcggtattc gccgttaata cgcaggccga
     2761 gggtttcgat acagatcgcg acacatttgt gggagcctat aactcgcttg gtgaagccgc
     2821 agttcccctt aagggagaaa gtgcaaatag cgttgcaagt ggatggtatc ctattggctc
     2881 tcatagcgtg gcagtgtctt tagcgcctgg agaatccaga gaattggttt atgtgcttgg
     2941 gtacgtggaa aatcctgatg aggagaagtg ggcagacgac gcaaaacagg tcgtgaataa
     3001 agaaagagct cacgctttat tatcaagatt tgctacgtcc gaacagactg acgcagcttt
     3061 cgccgctttg aaggattatt ggacggattt gttatcgacg tattccgtta gttccaatga
     3121 cgaaaaattg gaccggatgg ttaatatatg gaatcagtac caatgcatgg taacctttaa
     3181 catgagtcgc tcggcgagct ttttcgagac cggtattggg agagggatgg ggtttcgtga
     3241 tagcaaccaa gaccttctgg gctttgttca cttaatccca gagcgggcgc gtgagcgcat
     3301 cattgacata gcgtcaaccc agttcgcgga cggttccgct tatcaccaat accaaccttt
     3361 aacgaagcgc ggtaacaacg acattggctc tgggtttaac gacgatcctc tttggttaat
     3421 cgcgggaact gccgcctaca tcaaagaaac tggtgacttc tccatattag atgagcccgt
     3481 accttttgat aacgaaccgg ggtcagaagt tccgttattt gagcacctga cccgtagttt
     3541 tgagtttacc gtaacccatc ggggtcccca cggtttgcct cttattggcc gcgcagattg
     3601 gaacgattgt ttgaatctta attgcttcag tacaaccccg ggcgagtcat ttcagacaac
     3661 ggagaaccaa gccggtggcg tagcggagtc aacgttcata gccgcgcaat tcgttttata
     3721 tggggagcag tatgccgagt tggcagcgcg gcgcgggttg gccgacgtag ccgatagagc
     3781 aagagggcat gtggctgaaa tgagagacgc acttctgacc gacgggtggg acggcagctg
     3841 gttcttgaga gcttatgact attatggaaa ccctatcggc accgacgcac atgacgaagg
     3901 gaagatttgg atcgaaccgc aagggttcgc ggtgatggct ggagtcggtg taggcgaagg
     3961 gcctcaagat acggacgctc ccgctattaa ggcgcttgat tctgtgaacg aaatgttggc
     4021 aaccgatcac ggtatggtcc ttcaataccc agcatacact acataccaag tacacatggg
     4081 tgaagtatcc acttatccac ccgggtataa agagaatgga ggtatatttt gccacaataa
     4141 tccttgggtt atcatagcgg aaactgtcgt cggaagaggc ggacgggcat ttgattacta
     4201 caagagaatt actcccgcgt atcgcgagga cataagcgac gttcaccgtc ttgaacccta
     4261 tgtctatgca caaatgatag cagggaagga ggcggtccgt cacggtgaag caaagaacag
     4321 ctggcttacc gggactgcgg cttggaactt tgtgaccgtg tcacagtacc ttcttggggt
     4381 tcgcccagag tacgacggct tggtggtaga cccgcagatc ggacctgacg tacctagctt
     4441 tacggttacg cgtgtcgccc gtggagccac ttacgagata acagtcacta acagtgggac
     4501 cgacggatca cgcggtcgcc ttgtcgttga tggaacacct gtagaaggca acttggttcc
     4561 ttatgccccc gcagggtcaa cagttcgtgt cgatgtgaca ttgtagtgta cactagcata
     4621 accccttggg gcctctaaac gggtcttgag gggttttttg ctcgagtaaa aaaaatcctt
     4681 agctttcgct aaggatctgc agttaccaat gcttaatcag tgaggcacct atctcagcga
     4741 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     4801 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     4861 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     4921 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     4981 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     5041 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     5101 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     5161 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     5221 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     5281 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     5341 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     5401 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     5461 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     5521 caaaaaaggg aataagggcg acacggaaat gttgaatact catgcttaat ttctcctctt
     5581 taatagatct tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     5641 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgccagggtg gtttttcttt
     5701 tcaccagtga gacgggcaac agctgattgc ccttcaccgc ctggccctga gagagttgca
     5761 gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg
     5821 gcgggatata acatgagctg tcttcggtat cgtcgtatcc cactaccgag atatccgcac
     5881 caacgcgcag cccggactcg gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg
     5941 caaccagcat cgcagtggga acgatgccct cattcagcat ttgcatggtt tgttgaaaac
     6001 cggacatggc actccagtcg ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga
     6061 gatatttatg ccagccagcc agacgcagac gcgccgagac agaacttaat gggcccgcta
     6121 acagcgcgat ttgctggtga cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt
     6181 cttcatggga gaaaataata ctgttgatgg gtgtctggtc agagacatca agaaataacg
     6241 ccggaacatt agtgcaggca gcttccacag caatggcatc ctggtcatcc agcggatagt
     6301 taatgatcag cccactgacg cgttgcgcga gaagattgtg caccgccgct ttacaggctt
     6361 cgacgccgct tcgttctacc atcgacacca ccacgctggc acccagttga tcggcgcgag
     6421 atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc cagactggag gtggcaacgc
     6481 caatcagcaa cgactgtttg cccgccagtt gttgtgccac gcggttggga atgtaattca
     6541 gctccgccat cgccgcttcc actttttccc gcgttttcgc agaaacgtgg ctggcctggt
     6601 tcaccacgcg ggaaacggtc tgataagaga caccggcata ctctgcgaca tcgtataacg
     6661 ttactggttt cacattcacc accctgaatt gactctcttc cgggcgctat catgccatac
     6721 cgcgaaaggt tttgcaccat tcgatggtgt cgaattcatt ctcaccaata aaaaacgccc
     6781 ggcggcaacc gagcgttctg aacaaatcca gatggagttc tgaggtcatt actggatcta
     6841 tcaacaggag tcaagctttt acgcaacaac cggcggatta gcgatcaggt cgtcggagcg
     6901 atggtcacca gcactgtctt cccattccag cattgcgacc ggcgtggtat tatcaacacc
     6961 caggccgcgt ttcggttcga acgtcagggt ggcctgcgac gtttcaccac gccaggtaaa
     7021 ggaaatactc gtatcgtcat ccgtggtata tgagaaggtg ccgtcaaaag catccagttc
     7081 attgcgaaat ttcgccagag cgttcagtgc tttaaccacc ggacgtttca gattttcatc
     7141 gatttcggcc gtgctgtaat aatggcggtt aatgtcacga ccattgttgg ttttgcgcag
     7201 cagttccata tcatttttac ctgccagggc gcccacgtaa tagacttgcg gaacacccgg
     7261 caggaaaaac tgcaccgcac gcgcagcgat gtagtgctga tcgttacagc ccagagcgga
     7321 gtagtaggta ctattaactt ggtacagatc caggttcgat gccgccgcac cggtagctgc
     7381 ctgtgattcg ccgtgggtgt tcgcatgaat cgtattcacc aggttatcca catcttcgtc
     7441 cggcaccaga cctttcagcg agcggtccag ctgatctgaa ccgatatcaa taacgccgat
     7501 gccgtcatgc gtatccagga cggtaacagc attgttcgga cgaatatccg tccaatgggc
     7561 caccggttcg acgtggccgg tactcagggc atgcagcagc agcggcggca gggcaaagtc
     7621 ataaacgcga tccactttag acgcgatttc cacctgtttt ttgtaatagc tgtggacttc
     7681 gatcagaatt tccagaccgc gtttcacgcc ttcttcgcgc agacgggaaa tcagtttgaa
     7741 cgttttcggg gtcataaagc aactggtgcc tgcttctttg gcaccgtagc caaccgcatc
     7801 caggcggata taagacacat ggctggccgc catttggtcg aaaatggaca tcagatattc
     7861 ccaaccttta tcactgtccg tatcgatgtc cacttgctgc ggggtaaaag acacccagac
     7921 cagacgcgtt ttgcctgcga atttgtagtg ggtaaacggc agacccggac gcggacgata
     7981 aatgcctgcc aggtcttctt ccgtggcacc gttcgggaac acgctgctca tggtcagaaa
     8041 catcgggtaa tattctgatt cttcgccttt ggccagaaca tcttggaact gtttcgattc
     8101 ccatgacatg tgattaacga ttgcatccac cataatgttg tgggttttag acagttcggc
     8161 gacgtcatcc cagctaccca gacgttcatc aactttggta tggtcgatcg gatcgaaacc
     8221 tgcgtcggcg ccatcaaacg gggtgaaaaa cggcagaatg tggacaccgt cataaacgcc
     8281 atcaaagcgg gtacgcagga tgtccgtcat gcttttaatg gtgccatcac ccagacgatc
     8341 cgcataggta atcagttgga ctttattttt catggtaccc ttttcgaact gcgggtggct
     8401 ccagcttgc
//
