LOCUS       sequence-310538-        7105 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BaSP and CuCBP co-expression: medium RBS controls BaSP expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7105)
  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 7105)
  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: pBICI_medium
FEATURES             Location/Qualifiers
     source          1..7105
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     10..29
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        10..28
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    29..53
                     /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             69..89
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             109..126
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2636..2659
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     terminator      complement(4187..4234)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      4241..4335
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      4368..4956
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4857..4876
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5057..5848,5849..5917))
                     /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     5680..5699
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     RBS             5924..5935
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             complement(5945..7027)
                     /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     6989..7008
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(7028..7105)
                     /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 gaattcagtt aatacgactc actatagggg aattgtgagc ggataacaat tcccctctag
       61 aaataatgtt gtttaacttt aagaaggaga tatcatatgg gcagcagcca tcaccatcac
      121 catcacgggt cgacccgcta tggtcatttt gacgatgaag cacgcgagta cgttatcacc
      181 acaccgcata ctccgtaccc ttggatcaac tatctgggtt cggaacagtt cttcagtctt
      241 ttgtcgcacc aggctggcgg ctattcgttt tatcgcgatg ccaagatgcg tagattgaca
      301 agatacagat acaacaacat accagcagac gcgggaggcc ggtatcttta tgtaaacgac
      361 ggtggggacg tttggacgcc atcgtggctt cctgttaaag cagacttaga tcattttgaa
      421 gccagacatg gtcttggcta cagtaccatt acaggtgaaa gaaatggggt acgggttgag
      481 accttgtttt tcgtcccagt cggtgagaac gcggaggtcc agaaagttac ggtcactaat
      541 acctctgact cgtataaatc cctgacgttg ttttcttttg tcgaattttg tctttggaac
      601 gcgcaagacg atcaaactaa ctaccagcgt aatctgagca taggagaggt tgaagtggaa
      661 caagaatctc cgcatggcag tgcaatttat caccgtacag agtaccggga gagacgcgat
      721 cactacgcgg tattcgccgt taatacgcag gccgagggtt tcgatacaga tcgcgacaca
      781 tttgtgggag cctataactc gcttggtgaa gccgcagttc cccttaaggg agaaagtgca
      841 aatagcgttg caagtggatg gtatcctatt ggctctcata gcgtggcagt gtctttagcg
      901 cctggagaat ccagagaatt ggtttatgtg cttgggtacg tggaaaatcc tgatgaggag
      961 aagtgggcag acgacgcaaa acaggtcgtg aataaagaaa gagctcacgc tttattatca
     1021 agatttgcta cgtccgaaca gactgacgca gctttcgccg ctttgaagga ttattggacg
     1081 gatttgttat cgacgtattc cgttagttcc aatgacgaaa aattggaccg gatggttaat
     1141 atatggaatc agtaccaatg catggtaacc tttaacatga gtcgctcggc gagctttttc
     1201 gagaccggta ttgggagagg gatggggttt cgtgatagca accaagacct tctgggcttt
     1261 gttcacttaa tcccagagcg ggcgcgtgag cgcatcattg acatagcgtc aacccagttc
     1321 gcggacggtt ccgcttatca ccaataccaa cctttaacga agcgcggtaa caacgacatt
     1381 ggctctgggt ttaacgacga tcctctttgg ttaatcgcgg gaactgccgc ctacatcaaa
     1441 gaaactggtg acttctccat attagatgag cccgtacctt ttgataacga accggggtca
     1501 gaagttccgt tatttgagca cctgacccgt agttttgagt ttaccgtaac ccatcggggt
     1561 ccccacggtt tgcctcttat tggccgcgca gattggaacg attgtttgaa tcttaattgc
     1621 ttcagtacaa ccccgggcga gtcatttcag acaacggaga accaagccgg tggcgtagcg
     1681 gagtcaacgt tcatagccgc gcaattcgtt ttatatgggg agcagtatgc cgagttggca
     1741 gcgcggcgcg ggttggccga cgtagccgat agagcaagag ggcatgtggc tgaaatgaga
     1801 gacgcacttc tgaccgacgg gtgggacggc agctggttct tgagagctta tgactattat
     1861 ggaaacccta tcggcaccga cgcacatgac gaagggaaga tttggatcga accgcaaggg
     1921 ttcgcggtga tggctggagt cggtgtaggc gaagggcctc aagatacgga cgctcccgct
     1981 attaaggcgc ttgattctgt gaacgaaatg ttggcaaccg atcacggtat ggtccttcaa
     2041 tacccagcat acactacata ccaagtacac atgggtgaag tatccactta tccacccggg
     2101 tataaagaga atggaggtat attttgccac aataatcctt gggttatcat agcggaaact
     2161 gtcgtcggaa gaggcggacg ggcatttgat tactacaaga gaattactcc cgcgtatcgc
     2221 gaggacataa gcgacgttca ccgtcttgaa ccctatgtct atgcacaaat gatagcaggg
     2281 aaggaggcgg tccgtcacgg tgaagcaaag aacagctggc ttaccgggac tgcggcttgg
     2341 aactttgtga ccgtgtcaca gtaccttctt ggggttcgcc cagagtacga cggcttggtg
     2401 gtagacccgc agatcggacc tgacgtacct agctttacgg ttacgcgtgt cgcccgtgga
     2461 gccacttacg agataacagt cactaacagt gggaccgacg gatcacgcgg tcgccttgtc
     2521 gttgatggaa cacctgtaga aggcaacttg gttccttatg cccccgcagg gtcaacagtt
     2581 cgtgtcgatg tgacattgta gtgtacatca cacaggaaag acccccatgg caagctggag
     2641 ccacccgcag ttcgaaaagg gtaccatgaa aaataaagtc caactgatta cctatgcgga
     2701 tcgtctgggt gatggcacca ttaaaagcat gacggacatc ctgcgtaccc gctttgatgg
     2761 cgtttatgac ggtgtccaca ttctgccgtt tttcaccccg tttgatggcg ccgacgcagg
     2821 tttcgatccg atcgaccata ccaaagttga tgaacgtctg ggtagctggg atgacgtcgc
     2881 cgaactgtct aaaacccaca acattatggt ggatgcaatc gttaatcaca tgtcatggga
     2941 atcgaaacag ttccaagatg ttctggccaa aggcgaagaa tcagaatatt acccgatgtt
     3001 tctgaccatg agcagcgtgt tcccgaacgg tgccacggaa gaagacctgg caggcattta
     3061 tcgtccgcgt ccgggtctgc cgtttaccca ctacaaattc gcaggcaaaa cgcgtctggt
     3121 ctgggtgtct tttaccccgc agcaagtgga catcgatacg gacagtgata aaggttggga
     3181 atatctgatg tccattttcg accaaatggc ggccagccat gtgtcttata tccgcctgga
     3241 tgcggttggc tacggtgcca aagaagcagg caccagttgc tttatgaccc cgaaaacgtt
     3301 caaactgatt tcccgtctgc gcgaagaagg cgtgaaacgc ggtctggaaa ttctgatcga
     3361 agtccacagc tattacaaaa aacaggtgga aatcgcgtct aaagtggatc gcgtttatga
     3421 ctttgccctg ccgccgctgc tgctgcatgc cctgagtacc ggccacgtcg aaccggtggc
     3481 ccattggacg gatattcgtc cgaacaatgc tgttaccgtc ctggatacgc atgacggcat
     3541 cggcgttatt gatatcggtt cagatcagct ggaccgctcg ctgaaaggtc tggtgccgga
     3601 cgaagatgtg gataacctgg tgaatacgat tcatgcgaac acccacggcg aatcacaggc
     3661 agctaccggt gcggcggcat cgaacctgga tctgtaccaa gttaatagta cctactactc
     3721 cgctctgggc tgtaacgatc agcactacat cgctgcgcgt gcggtgcagt ttttcctgcc
     3781 gggtgttccg caagtctatt acgtgggcgc cctggcaggt aaaaatgata tggaactgct
     3841 gcgcaaaacc aacaatggtc gtgacattaa ccgccattat tacagcacgg ccgaaatcga
     3901 tgaaaatctg aaacgtccgg tggttaaagc actgaacgct ctggcgaaat ttcgcaatga
     3961 actggatgct tttgacggca ccttctcata taccacggat gacgatacga gtatttcctt
     4021 tacctggcgt ggtgaaacgt cgcaggccac cctgacgttc gaaccgaaac gcggcctggg
     4081 tgttgataat accacgccgg tcgcaatgct ggaatgggaa gacagtgctg gtgaccatcg
     4141 ctccgacgac ctgatcgcta atccgccggt tgttgcgtaa aagcttcaaa aaacccctca
     4201 agacccgttt agaggcccca aggggttatg ctaggagctc gactcctgtt gatagatcca
     4261 gtaatgacct cagaactcca tctggatttg ttcagaacgc tcggttgccg ccgggcgttt
     4321 tttattggtg agaatggatc ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     4381 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     4441 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     4501 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     4561 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     4621 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     4681 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     4741 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     4801 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     4861 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     4921 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     4981 cggttcctgg ccttttgctg gccttttgct cacactcgag taaaaaaaat ccttagcttt
     5041 cgctaaggat ctgcagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     5101 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     5161 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     5221 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     5281 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     5341 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     5401 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     5461 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     5521 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     5581 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     5641 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     5701 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     5761 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     5821 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     5881 agggaataag ggcgacacgg aaatgttgaa tactcatgct taatttctcc tctttaatag
     5941 atcttcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     6001 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgccag ggtggttttt cttttcacca
     6061 gtgagacggg caacagctga ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc
     6121 ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt aacggcggga
     6181 tataacatga gctgtcctcg gtatcgtcgt atcccactac cgagatatcc gcaccaacgc
     6241 gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg ttggcaacca
     6301 gcatcgcagt gggaacgatg ccctcattca gcatttgcat ggtttgttga aaaccggaca
     6361 tggcactcca gtcgccttcc cgttccgcta tcggctgaat ttgattgcga gtgagatatt
     6421 tatgccagcc agccagacgc agacgcgccg agacagaact taatgggccc gctaacagcg
     6481 cgatttgctg gtgacccaat gcgaccagat gctccacgcc cagtcgcgta ccgtcctcat
     6541 gggagaaaat aatactgttg atgggtgtct ggtcagagac atcaagaaat aacgccggaa
     6601 cattagtgca ggcagcttcc acagcaatgg catcctggtc atccagcgga tagttaatga
     6661 tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc
     6721 cgcttcgttc taccatcgac accaccacgc tggcacccag ttgatcggcg cgagatttaa
     6781 tcgccgcgac aatttgcgac ggcgcgtgca gggccagact ggaggtggca acgccaatca
     6841 gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg
     6901 ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac gtggctggcc tggttcacca
     6961 cgcgggaaac ggtctgataa gagacaccgg catactctgc gacatcgtat aacgttactg
     7021 gtttcacatt caccaccctg aattgactct cttccgggcg ctatcatgcc ataccgcgaa
     7081 aggttttgca ccattcgatg gtgtc
//
