LOCUS       sequence-310535-        7103 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BaSP and CuCBP co-expression: weak RBS controls BaSP expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7103)
  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 7103)
  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_weak
FEATURES             Location/Qualifiers
     source          1..7103
                     /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             2634..2657
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     terminator      complement(4185..4232)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      4239..4333
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      4366..4954
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4855..4874
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5055..5846,5847..5915))
                     /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     5678..5697
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     RBS             5922..5933
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             complement(5943..7025)
                     /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     6987..7006
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(7026..7103)
                     /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 cacaggacac ccccatggca agctggagcc
     2641 acccgcagtt cgaaaagggt accatgaaaa ataaagtcca actgattacc tatgcggatc
     2701 gtctgggtga tggcaccatt aaaagcatga cggacatcct gcgtacccgc tttgatggcg
     2761 tttatgacgg tgtccacatt ctgccgtttt tcaccccgtt tgatggcgcc gacgcaggtt
     2821 tcgatccgat cgaccatacc aaagttgatg aacgtctggg tagctgggat gacgtcgccg
     2881 aactgtctaa aacccacaac attatggtgg atgcaatcgt taatcacatg tcatgggaat
     2941 cgaaacagtt ccaagatgtt ctggccaaag gcgaagaatc agaatattac ccgatgtttc
     3001 tgaccatgag cagcgtgttc ccgaacggtg ccacggaaga agacctggca ggcatttatc
     3061 gtccgcgtcc gggtctgccg tttacccact acaaattcgc aggcaaaacg cgtctggtct
     3121 gggtgtcttt taccccgcag caagtggaca tcgatacgga cagtgataaa ggttgggaat
     3181 atctgatgtc cattttcgac caaatggcgg ccagccatgt gtcttatatc cgcctggatg
     3241 cggttggcta cggtgccaaa gaagcaggca ccagttgctt tatgaccccg aaaacgttca
     3301 aactgatttc ccgtctgcgc gaagaaggcg tgaaacgcgg tctggaaatt ctgatcgaag
     3361 tccacagcta ttacaaaaaa caggtggaaa tcgcgtctaa agtggatcgc gtttatgact
     3421 ttgccctgcc gccgctgctg ctgcatgccc tgagtaccgg ccacgtcgaa ccggtggccc
     3481 attggacgga tattcgtccg aacaatgctg ttaccgtcct ggatacgcat gacggcatcg
     3541 gcgttattga tatcggttca gatcagctgg accgctcgct gaaaggtctg gtgccggacg
     3601 aagatgtgga taacctggtg aatacgattc atgcgaacac ccacggcgaa tcacaggcag
     3661 ctaccggtgc ggcggcatcg aacctggatc tgtaccaagt taatagtacc tactactccg
     3721 ctctgggctg taacgatcag cactacatcg ctgcgcgtgc ggtgcagttt ttcctgccgg
     3781 gtgttccgca agtctattac gtgggcgccc tggcaggtaa aaatgatatg gaactgctgc
     3841 gcaaaaccaa caatggtcgt gacattaacc gccattatta cagcacggcc gaaatcgatg
     3901 aaaatctgaa acgtccggtg gttaaagcac tgaacgctct ggcgaaattt cgcaatgaac
     3961 tggatgcttt tgacggcacc ttctcatata ccacggatga cgatacgagt atttccttta
     4021 cctggcgtgg tgaaacgtcg caggccaccc tgacgttcga accgaaacgc ggcctgggtg
     4081 ttgataatac cacgccggtc gcaatgctgg aatgggaaga cagtgctggt gaccatcgct
     4141 ccgacgacct gatcgctaat ccgccggttg ttgcgtaaaa gcttcaaaaa acccctcaag
     4201 acccgtttag aggccccaag gggttatgct aggagctcga ctcctgttga tagatccagt
     4261 aatgacctca gaactccatc tggatttgtt cagaacgctc ggttgccgcc gggcgttttt
     4321 tattggtgag aatggatccc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     4381 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     4441 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     4501 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     4561 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     4621 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     4681 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     4741 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     4801 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     4861 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     4921 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     4981 gttcctggcc ttttgctggc cttttgctca cactcgagta aaaaaaatcc ttagctttcg
     5041 ctaaggatct gcagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     5101 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     5161 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     5221 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     5281 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     5341 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     5401 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     5461 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     5521 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     5581 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     5641 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     5701 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     5761 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     5821 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     5881 ggaataaggg cgacacggaa atgttgaata ctcatgctta atttctcctc tttaatagat
     5941 cttcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     6001 aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg tggtttttct tttcaccagt
     6061 gagacgggca acagctgatt gcccttcacc gcctggccct gagagagttg cagcaagcgg
     6121 tccacgctgg tttgccccag caggcgaaaa tcctgtttga tggtggttaa cggcgggata
     6181 taacatgagc tgtcctcggt atcgtcgtat cccactaccg agatatccgc accaacgcgc
     6241 agcccggact cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc
     6301 atcgcagtgg gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa accggacatg
     6361 gcactccagt cgccttcccg ttccgctatc ggctgaattt gattgcgagt gagatattta
     6421 tgccagccag ccagacgcag acgcgccgag acagaactta atgggcccgc taacagcgcg
     6481 atttgctggt gacccaatgc gaccagatgc tccacgccca gtcgcgtacc gtcctcatgg
     6541 gagaaaataa tactgttgat gggtgtctgg tcagagacat caagaaataa cgccggaaca
     6601 ttagtgcagg cagcttccac agcaatggca tcctggtcat ccagcggata gttaatgatc
     6661 agcccactga cgcgttgcgc gagaagattg tgcaccgccg ctttacaggc ttcgacgccg
     6721 cttcgttcta ccatcgacac caccacgctg gcacccagtt gatcggcgcg agatttaatc
     6781 gccgcgacaa tttgcgacgg cgcgtgcagg gccagactgg aggtggcaac gccaatcagc
     6841 aacgactgtt tgcccgccag ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc
     6901 atcgccgctt ccactttttc ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg
     6961 cgggaaacgg tctgataaga gacaccggca tactctgcga catcgtataa cgttactggt
     7021 ttcacattca ccaccctgaa ttgactctct tccgggcgct atcatgccat accgcgaaag
     7081 gttttgcacc attcgatggt gtc
//
