LOCUS       sequence-310554-        7187 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BaSP and CuCBP co-expression: lacZ promoter controls BaSP 
            expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7187)
  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 7187)
  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_lacZ
FEATURES             Location/Qualifiers
     source          1..7187
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(7..101)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1629..1652)
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(1672..1694)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1683..1699
                     /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)."
     promoter        complement(join(1707..1713,1714..1731,1732..1737))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     rep_origin      1784..2372
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2273..2292
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2451..2470
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2451..2469
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2470..2494
                     /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             2509..2531
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2550..2567
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      5049..5096
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(join(5139..5930,5931..5999))
                     /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     5762..5781
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     RBS             6006..6017
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             complement(6027..7109)
                     /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     7071..7090
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(7110..7187)
                     /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."
ORIGIN
        1 gaattcattc tcaccaataa aaaacgcccg gcggcaaccg agcgttctga acaaatccag
       61 atggagttct gaggtcatta ctggatctat caacaggagt caagctttta cgcaacaacc
      121 ggcggattag cgatcaggtc gtcggagcga tggtcaccag cactgtcttc ccattccagc
      181 attgcgaccg gcgtggtatt atcaacaccc aggccgcgtt tcggttcgaa cgtcagggtg
      241 gcctgcgacg tttcaccacg ccaggtaaag gaaatactcg tatcgtcatc cgtggtatat
      301 gagaaggtgc cgtcaaaagc atccagttca ttgcgaaatt tcgccagagc gttcagtgct
      361 ttaaccaccg gacgtttcag attttcatcg atttcggccg tgctgtaata atggcggtta
      421 atgtcacgac cattgttggt tttgcgcagc agttccatat catttttacc tgccagggcg
      481 cccacgtaat agacttgcgg aacacccggc aggaaaaact gcaccgcacg cgcagcgatg
      541 tagtgctgat cgttacagcc cagagcggag tagtaggtac tattaacttg gtacagatcc
      601 aggttcgatg ccgccgcacc ggtagctgcc tgtgattcgc cgtgggtgtt cgcatgaatc
      661 gtattcacca ggttatccac atcttcgtcc ggcaccagac ctttcagcga gcggtccagc
      721 tgatctgaac cgatatcaat aacgccgatg ccgtcatgcg tatccaggac ggtaacagca
      781 ttgttcggac gaatatccgt ccaatgggcc accggttcga cgtggccggt actcagggca
      841 tgcagcagca gcggcggcag ggcaaagtca taaacgcgat ccactttaga cgcgatttcc
      901 acctgttttt tgtaatagct gtggacttcg atcagaattt ccagaccgcg tttcacgcct
      961 tcttcgcgca gacgggaaat cagtttgaac gttttcgggg tcataaagca actggtgcct
     1021 gcttctttgg caccgtagcc aaccgcatcc aggcggatat aagacacatg gctggccgcc
     1081 atttggtcga aaatggacat cagatattcc caacctttat cactgtccgt atcgatgtcc
     1141 acttgctgcg gggtaaaaga cacccagacc agacgcgttt tgcctgcgaa tttgtagtgg
     1201 gtaaacggca gacccggacg cggacgataa atgcctgcca ggtcttcttc cgtggcaccg
     1261 ttcgggaaca cgctgctcat ggtcagaaac atcgggtaat attctgattc ttcgcctttg
     1321 gccagaacat cttggaactg tttcgattcc catgacatgt gattaacgat tgcatccacc
     1381 ataatgttgt gggttttaga cagttcggcg acgtcatccc agctacccag acgttcatca
     1441 actttggtat ggtcgatcgg atcgaaacct gcgtcggcgc catcaaacgg ggtgaaaaac
     1501 ggcagaatgt ggacaccgtc ataaacgcca tcaaagcggg tacgcaggat gtccgtcatg
     1561 cttttaatgg tgccatcacc cagacgatcc gcataggtaa tcagttggac tttatttttc
     1621 atggtaccct tttcgaactg cgggtggctc cagcttgcca tggagctgtt tcctgtgtga
     1681 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     1741 tggggtgcct aggatccccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     1801 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     1861 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     1921 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     1981 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     2041 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     2101 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     2161 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     2221 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     2281 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     2341 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     2401 tcctggcctt ttgctggcct tttgctcaca actagtcgat cccgcgaaat taatacgact
     2461 cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt tgtttaactt
     2521 taagaaggag atatcatatg ggcagcagcc atcaccatca ccatcacggg tcgacccgct
     2581 atggtcattt tgacgatgaa gcacgcgagt acgttatcac cacaccgcat actccgtacc
     2641 cttggatcaa ctatctgggt tcggaacagt tcttcagtct tttgtcgcac caggctggcg
     2701 gctattcgtt ttatcgcgat gccaagatgc gtagattgac aagatacaga tacaacaaca
     2761 taccagcaga cgcgggaggc cggtatcttt atgtaaacga cggtggggac gtttggacgc
     2821 catcgtggct tcctgttaaa gcagacttag atcattttga agccagacat ggtcttggct
     2881 acagtaccat tacaggtgaa agaaatgggg tacgggttga gaccttgttt ttcgtcccag
     2941 tcggtgagaa cgcggaggtc cagaaagtta cggtcactaa tacctctgac tcgtataaat
     3001 ccctgacgtt gttttctttt gtcgaatttt gtctttggaa cgcgcaagac gatcaaacta
     3061 actaccagcg taatctgagc ataggagagg ttgaagtgga acaagaatct ccgcatggca
     3121 gtgcaattta tcaccgtaca gagtaccggg agagacgcga tcactacgcg gtattcgccg
     3181 ttaatacgca ggccgagggt ttcgatacag atcgcgacac atttgtggga gcctataact
     3241 cgcttggtga agccgcagtt ccccttaagg gagaaagtgc aaatagcgtt gcaagtggat
     3301 ggtatcctat tggctctcat agcgtggcag tgtctttagc gcctggagaa tccagagaat
     3361 tggtttatgt gcttgggtac gtggaaaatc ctgatgagga gaagtgggca gacgacgcaa
     3421 aacaggtcgt gaataaagaa agagctcacg ctttattatc aagatttgct acgtccgaac
     3481 agactgacgc agctttcgcc gctttgaagg attattggac ggatttgtta tcgacgtatt
     3541 ccgttagttc caatgacgaa aaattggacc ggatggttaa tatatggaat cagtaccaat
     3601 gcatggtaac ctttaacatg agtcgctcgg cgagcttttt cgagaccggt attgggagag
     3661 ggatggggtt tcgtgatagc aaccaagacc ttctgggctt tgttcactta atcccagagc
     3721 gggcgcgtga gcgcatcatt gacatagcgt caacccagtt cgcggacggt tccgcttatc
     3781 accaatacca acctttaacg aagcgcggta acaacgacat tggctctggg tttaacgacg
     3841 atcctctttg gttaatcgcg ggaactgccg cctacatcaa agaaactggt gacttctcca
     3901 tattagatga gcccgtacct tttgataacg aaccggggtc agaagttccg ttatttgagc
     3961 acctgacccg tagttttgag tttaccgtaa cccatcgggg tccccacggt ttgcctctta
     4021 ttggccgcgc agattggaac gattgtttga atcttaattg cttcagtaca accccgggcg
     4081 agtcatttca gacaacggag aaccaagccg gtggcgtagc ggagtcaacg ttcatagccg
     4141 cgcaattcgt tttatatggg gagcagtatg ccgagttggc agcgcggcgc gggttggccg
     4201 acgtagccga tagagcaaga gggcatgtgg ctgaaatgag agacgcactt ctgaccgacg
     4261 ggtgggacgg cagctggttc ttgagagctt atgactatta tggaaaccct atcggcaccg
     4321 acgcacatga cgaagggaag atttggatcg aaccgcaagg gttcgcggtg atggctggag
     4381 tcggtgtagg cgaagggcct caagatacgg acgctcccgc tattaaggcg cttgattctg
     4441 tgaacgaaat gttggcaacc gatcacggta tggtccttca atacccagca tacactacat
     4501 accaagtaca catgggtgaa gtatccactt atccacccgg gtataaagag aatggaggta
     4561 tattttgcca caataatcct tgggttatca tagcggaaac tgtcgtcgga agaggcggac
     4621 gggcatttga ttactacaag agaattactc ccgcgtatcg cgaggacata agcgacgttc
     4681 accgtcttga accctatgtc tatgcacaaa tgatagcagg gaaggaggcg gtccgtcacg
     4741 gtgaagcaaa gaacagctgg cttaccggga ctgcggcttg gaactttgtg accgtgtcac
     4801 agtaccttct tggggttcgc ccagagtacg acggcttggt ggtagacccg cagatcggac
     4861 ctgacgtacc tagctttacg gttacgcgtg tcgcccgtgg agccacttac gagataacag
     4921 tcactaacag tgggaccgac ggatcacgcg gtcgccttgt cgttgatgga acacctgtag
     4981 aaggcaactt ggttccttat gcccccgcag ggtcaacagt tcgtgtcgat gtgacattgt
     5041 agtgtacact agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctcg
     5101 agtaaaaaaa atccttagct ttcgctaagg atctgcagtt accaatgctt aatcagtgag
     5161 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     5221 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     5281 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     5341 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     5401 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     5461 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     5521 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     5581 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     5641 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     5701 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     5761 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     5821 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     5881 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     5941 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcatg
     6001 cttaatttct cctctttaat agatcttcac tgcccgcttt ccagtcggga aacctgtcgt
     6061 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgcc
     6121 agggtggttt ttcttttcac cagtgagacg ggcaacagct gattgccctt caccgcctgg
     6181 ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt
     6241 ttgatggtgg ttaacggcgg gatataacat gagctgtcct cggtatcgtc gtatcccact
     6301 accgagatat ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc
     6361 gccatctgat cgttggcaac cagcatcgca gtgggaacga tgccctcatt cagcatttgc
     6421 atggtttgtt gaaaaccgga catggcactc cagtcgcctt cccgttccgc tatcggctga
     6481 atttgattgc gagtgagata tttatgccag ccagccagac gcagacgcgc cgagacagaa
     6541 cttaatgggc ccgctaacag cgcgatttgc tggtgaccca atgcgaccag atgctccacg
     6601 cccagtcgcg taccgtcctc atgggagaaa ataatactgt tgatgggtgt ctggtcagag
     6661 acatcaagaa ataacgccgg aacattagtg caggcagctt ccacagcaat ggcatcctgg
     6721 tcatccagcg gatagttaat gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc
     6781 gccgctttac aggcttcgac gccgcttcgt tctaccatcg acaccaccac gctggcaccc
     6841 agttgatcgg cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg cagggccaga
     6901 ctggaggtgg caacgccaat cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg
     6961 ttgggaatgt aattcagctc cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa
     7021 acgtggctgg cctggttcac cacgcgggaa acggtctgat aagagacacc ggcatactct
     7081 gcgacatcgt ataacgttac tggtttcaca ttcaccaccc tgaattgact ctcttccggg
     7141 cgctatcatg ccataccgcg aaaggttttg caccattcga tggtgtc
//
