LOCUS       sequence-311675-        7211 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pBICI_2_strong without bom site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7211)
  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 7211)
  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: pBR322-bom
FEATURES             Location/Qualifiers
     source          1..7211
                     /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"
     RBS             2608..2619
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             2635..2658
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     terminator      complement(4186..4233)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      4240..4334
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(4345..4362)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4516..5104)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4596..4615)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5163..5954,5955..6023))
                     /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     5786..5805
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     RBS             6030..6041
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             complement(6051..7133)
                     /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     7095..7114
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(7134..7211)
                     /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 gtgtacaaaa gaggagaaaa cccccatggc aagctggagc
     2641 cacccgcagt tcgaaaaggg taccatgaaa aataaagtcc aactgattac ctatgcggat
     2701 cgtctgggtg atggcaccat taaaagcatg acggacatcc tgcgtacccg ctttgatggc
     2761 gtttatgacg gtgtccacat tctgccgttt ttcaccccgt ttgatggcgc cgacgcaggt
     2821 ttcgatccga tcgaccatac caaagttgat gaacgtctgg gtagctggga tgacgtcgcc
     2881 gaactgtcta aaacccacaa cattatggtg gatgcaatcg ttaatcacat gtcatgggaa
     2941 tcgaaacagt tccaagatgt tctggccaaa ggcgaagaat cagaatatta cccgatgttt
     3001 ctgaccatga gcagcgtgtt cccgaacggt gccacggaag aagacctggc aggcatttat
     3061 cgtccgcgtc cgggtctgcc gtttacccac tacaaattcg caggcaaaac gcgtctggtc
     3121 tgggtgtctt ttaccccgca gcaagtggac atcgatacgg acagtgataa aggttgggaa
     3181 tatctgatgt ccattttcga ccaaatggcg gccagccatg tgtcttatat ccgcctggat
     3241 gcggttggct acggtgccaa agaagcaggc accagttgct ttatgacccc gaaaacgttc
     3301 aaactgattt cccgtctgcg cgaagaaggc gtgaaacgcg gtctggaaat tctgatcgaa
     3361 gtccacagct attacaaaaa acaggtggaa atcgcgtcta aagtggatcg cgtttatgac
     3421 tttgccctgc cgccgctgct gctgcatgcc ctgagtaccg gccacgtcga accggtggcc
     3481 cattggacgg atattcgtcc gaacaatgct gttaccgtcc tggatacgca tgacggcatc
     3541 ggcgttattg atatcggttc agatcagctg gaccgctcgc tgaaaggtct ggtgccggac
     3601 gaagatgtgg ataacctggt gaatacgatt catgcgaaca cccacggcga atcacaggca
     3661 gctaccggtg cggcggcatc gaacctggat ctgtaccaag ttaatagtac ctactactcc
     3721 gctctgggct gtaacgatca gcactacatc gctgcgcgtg cggtgcagtt tttcctgccg
     3781 ggtgttccgc aagtctatta cgtgggcgcc ctggcaggta aaaatgatat ggaactgctg
     3841 cgcaaaacca acaatggtcg tgacattaac cgccattatt acagcacggc cgaaatcgat
     3901 gaaaatctga aacgtccggt ggttaaagca ctgaacgctc tggcgaaatt tcgcaatgaa
     3961 ctggatgctt ttgacggcac cttctcatat accacggatg acgatacgag tatttccttt
     4021 acctggcgtg gtgaaacgtc gcaggccacc ctgacgttcg aaccgaaacg cggcctgggt
     4081 gttgataata ccacgccggt cgcaatgctg gaatgggaag acagtgctgg tgaccatcgc
     4141 tccgacgacc tgatcgctaa tccgccggtt gttgcgtaaa agcttcaaaa aacccctcaa
     4201 gacccgttta gaggccccaa ggggttatgc taggagctcg actcctgttg atagatccag
     4261 taatgacctc agaactccat ctggatttgt tcagaacgct cggttgccgc cgggcgtttt
     4321 ttattggtga gaatggatcg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     4381 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     4441 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     4501 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     4561 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     4621 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     4681 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     4741 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     4801 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     4861 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     4921 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
     4981 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     5041 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     5101 tcaagaagat cctttgatct ctcgagtaaa aaaaatcctt agctttcgct aaggatctgc
     5161 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     5221 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     5281 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     5341 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     5401 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     5461 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     5521 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     5581 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     5641 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     5701 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     5761 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     5821 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     5881 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     5941 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     6001 acacggaaat gttgaatact catgcttaat ttctcctctt taatagatct tcactgcccg
     6061 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     6121 gaggcggttt gcgtattggg cgccagggtg gtttttcttt tcaccagtga gacgggcaac
     6181 agctgattgc ccttcaccgc ctggccctga gagagttgca gcaagcggtc cacgctggtt
     6241 tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata acatgagctg
     6301 tcctcggtat cgtcgtatcc cactaccgag atatccgcac caacgcgcag cccggactcg
     6361 gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg caaccagcat cgcagtggga
     6421 acgatgccct cattcagcat ttgcatggtt tgttgaaaac cggacatggc actccagtcg
     6481 ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga gatatttatg ccagccagcc
     6541 agacgcagac gcgccgagac agaacttaat gggcccgcta acagcgcgat ttgctggtga
     6601 cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt cctcatggga gaaaataata
     6661 ctgttgatgg gtgtctggtc agagacatca agaaataacg ccggaacatt agtgcaggca
     6721 gcttccacag caatggcatc ctggtcatcc agcggatagt taatgatcag cccactgacg
     6781 cgttgcgcga gaagattgtg caccgccgct ttacaggctt cgacgccgct tcgttctacc
     6841 atcgacacca ccacgctggc acccagttga tcggcgcgag atttaatcgc cgcgacaatt
     6901 tgcgacggcg cgtgcagggc cagactggag gtggcaacgc caatcagcaa cgactgtttg
     6961 cccgccagtt gttgtgccac gcggttggga atgtaattca gctccgccat cgccgcttcc
     7021 actttttccc gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc
     7081 tgataagaga caccggcata ctctgcgaca tcgtataacg ttactggttt cacattcacc
     7141 accctgaatt gactctcttc cgggcgctat catgccatac cgcgaaaggt tttgcaccat
     7201 tcgatggtgt c
//
