LOCUS       sequence-310557-        7462 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  see pDUAL_lacZ: exchanged origin of replication (pBR322) and 
            introduced bom site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7462)
  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 7462)
  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_2_lacZ
FEATURES             Location/Qualifiers
     source          1..7462
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(20..25,26..43,44..50)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    58..74
                     /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)."
     primer_bind     63..85
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             105..128
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     terminator      1656..1750
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     promoter        1757..1834
                     /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."
     CDS             1835..2917
                     /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
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVE*SGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(1854..1873)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     RBS             2927..2938
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             join(2945..3013,3014..3805)
                     /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     complement(3163..3182)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      complement(3848..3895)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(6377..6394)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             6413..6435
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    6450..6474
                     /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)."
     primer_bind     complement(6474..6493)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6475..6493)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      6530..7118
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7019..7038
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7272..7289
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    7304..7444
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(7370..7389)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gatcctaggc accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg
       61 tgagcggata acaatttcac acaggaaaca gctccatggc aagctggagc cacccgcagt
      121 tcgaaaaggg taccatgaaa aataaagtcc aactgattac ctatgcggat cgtctgggtg
      181 atggcaccat taaaagcatg acggacatcc tgcgtacccg ctttgatggc gtttatgacg
      241 gtgtccacat tctgccgttt ttcaccccgt ttgatggcgc cgacgcaggt ttcgatccga
      301 tcgaccatac caaagttgat gaacgtctgg gtagctggga tgacgtcgcc gaactgtcta
      361 aaacccacaa cattatggtg gatgcaatcg ttaatcacat gtcatgggaa tcgaaacagt
      421 tccaagatgt tctggccaaa ggcgaagaat cagaatatta cccgatgttt ctgaccatga
      481 gcagcgtgtt cccgaacggt gccacggaag aagacctggc aggcatttat cgtccgcgtc
      541 cgggtctgcc gtttacccac tacaaattcg caggcaaaac gcgtctggtc tgggtgtctt
      601 ttaccccgca gcaagtggac atcgatacgg acagtgataa aggttgggaa tatctgatgt
      661 ccattttcga ccaaatggcg gccagccatg tgtcttatat ccgcctggat gcggttggct
      721 acggtgccaa agaagcaggc accagttgct ttatgacccc gaaaacgttc aaactgattt
      781 cccgtctgcg cgaagaaggc gtgaaacgcg gtctggaaat tctgatcgaa gtccacagct
      841 attacaaaaa acaggtggaa atcgcgtcta aagtggatcg cgtttatgac tttgccctgc
      901 cgccgctgct gctgcatgcc ctgagtaccg gccacgtcga accggtggcc cattggacgg
      961 atattcgtcc gaacaatgct gttaccgtcc tggatacgca tgacggcatc ggcgttattg
     1021 atatcggttc agatcagctg gaccgctcgc tgaaaggtct ggtgccggac gaagatgtgg
     1081 ataacctggt gaatacgatt catgcgaaca cccacggcga atcacaggca gctaccggtg
     1141 cggcggcatc gaacctggat ctgtaccaag ttaatagtac ctactactcc gctctgggct
     1201 gtaacgatca gcactacatc gctgcgcgtg cggtgcagtt tttcctgccg ggtgttccgc
     1261 aagtctatta cgtgggcgcc ctggcaggta aaaatgatat ggaactgctg cgcaaaacca
     1321 acaatggtcg tgacattaac cgccattatt acagcacggc cgaaatcgat gaaaatctga
     1381 aacgtccggt ggttaaagca ctgaacgctc tggcgaaatt tcgcaatgaa ctggatgctt
     1441 ttgacggcac cttctcatat accacggatg acgatacgag tatttccttt acctggcgtg
     1501 gtgaaacgtc gcaggccacc ctgacgttcg aaccgaaacg cggcctgggt gttgataata
     1561 ccacgccggt cgcaatgctg gaatgggaag acagtgctgg tgaccatcgc tccgacgacc
     1621 tgatcgctaa tccgccggtt gttgcgtaaa agcttgactc ctgttgatag atccagtaat
     1681 gacctcagaa ctccatctgg atttgttcag aacgctcggt tgccgccggg cgttttttat
     1741 tggtgagaat gaattcgaca ccatcgaatg gtgcaaaacc tttcgcggta tggcatgata
     1801 gcgcccggaa gagagtcaat tcagggtggt gaatgtgaaa ccagtaacgt tatacgatgt
     1861 cgcagagtat gccggtgtct cttatcagac cgtttcccgc gtggtgaacc aggccagcca
     1921 cgtttctgcg aaaacgcggg aaaaagtgga agcggcgatg gcggagctga attacattcc
     1981 caaccgcgtg gcacaacaac tggcgggcaa acagtcgttg ctgattggcg ttgccacctc
     2041 cagtctggcc ctgcacgcgc cgtcgcaaat tgtcgcggcg attaaatctc gcgccgatca
     2101 actgggtgcc agcgtggtgg tgtcgatggt agaatgaagc ggcgtcgaag cctgtaaagc
     2161 ggcggtgcac aatcttctcg cgcaacgcgt cagtgggctg atcattaact atccgctgga
     2221 tgaccaggat gccattgctg tggaagctgc ctgcactaat gttccggcgt tatttcttga
     2281 tgtctctgac cagacaccca tcaacagtat tattttctcc catgaggacg gtacgcgact
     2341 gggcgtggag catctggtcg cattgggtca ccagcaaatc gcgctgttag cgggcccatt
     2401 aagttctgtc tcggcgcgtc tgcgtctggc tggctggcat aaatatctca ctcgcaatca
     2461 aattcagccg atagcggaac gggaaggcga ctggagtgcc atgtccggtt ttcaacaaac
     2521 catgcaaatg ctgaatgagg gcatcgttcc cactgcgatg ctggttgcca acgatcagat
     2581 ggcgctgggc gcaatgcgcg ccattaccga gtccgggctg cgcgttggtg cggatatctc
     2641 ggtagtggga tacgacgata ccgaggacag ctcatgttat atcccgccgt taaccaccat
     2701 caaacaggat tttcgcctgc tggggcaaac cagcgtggac cgcttgctgc aactctctca
     2761 gggccaggcg gtgaagggca atcagctgtt gcccgtctca ctggtgaaaa gaaaaaccac
     2821 cctggcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     2881 ggcacgacag gtttcccgac tggaaagcgg gcagtgaaga tctattaaag aggagaaatt
     2941 aagcatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     3001 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     3061 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     3121 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     3181 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     3241 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     3301 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     3361 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     3421 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     3481 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     3541 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     3601 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     3661 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     3721 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     3781 tgcctcactg attaagcatt ggtaactgca gatccttagc gaaagctaag gatttttttt
     3841 actcgagcaa aaaacccctc aagacccgtt tagaggcccc aaggggttat gctagtgtac
     3901 actacaatgt cacatcgaca cgaactgttg accctgcggg ggcataagga accaagttgc
     3961 cttctacagg tgttccatca acgacaaggc gaccgcgtga tccgtcggtc ccactgttag
     4021 tgactgttat ctcgtaagtg gctccacggg cgacacgcgt aaccgtaaag ctaggtacgt
     4081 caggtccgat ctgcgggtct accaccaagc cgtcgtactc tgggcgaacc ccaagaaggt
     4141 actgtgacac ggtcacaaag ttccaagccg cagtcccggt aagccagctg ttctttgctt
     4201 caccgtgacg gaccgcctcc ttccctgcta tcatttgtgc atagacatag ggttcaagac
     4261 ggtgaacgtc gcttatgtcc tcgcgatacg cgggagtaat tctcttgtag taatcaaatg
     4321 cccgtccgcc tcttccgacg acagtttccg ctatgataac ccaaggatta ttgtggcaaa
     4381 atatacctcc attctcttta tacccgggtg gataagtgga tacttcaccc atgtgtactt
     4441 ggtatgtagt gtatgctggg tattgaagga ccataccgtg atcggttgcc aacatttcgt
     4501 tcacagaatc aagcgcctta atagcgggag cgtccgtatc ttgaggccct tcgcctacac
     4561 cgactccagc catcaccgcg aacccttgcg gttcgatcca aatcttccct tcgtcatgtg
     4621 cgtcggtgcc gatagggttt ccataatagt cataagctct caagaaccag ctgccgtccc
     4681 acccgtcggt cagaagtgcg tctctcattt cagccacatg ccctcttgct ctatcggcta
     4741 cgtcggccaa cccgcgccgc gctgccaact cggcatactg ctccccatat aaaacgaatt
     4801 gcgcggctat gaacgttgac tccgctacgc caccggcttg gttctccgtt gtctgaaatg
     4861 actcgcccgg ggttgtactg aagcaattaa gattcaaaca atcgttccaa tctgcgcggc
     4921 caataagagg caaaccgtgg ggaccccgat gggttacggt aaactcaaaa ctacgggtca
     4981 ggtgctcaaa taacggaact tctgaccccg gttcgttatc aaaaggtacg ggctcatcta
     5041 atatggagaa gtcaccagtt tctttgatgt aggcggcagt tcccgcgatt aaccaaagag
     5101 gatcgtcgtt aaacccagag ccaatgtcgt tgttaccgcg cttcgttaaa ggttggtatt
     5161 ggtgataagc ggaaccgtcc gcgaactggg ttgacgctat gtcaatgatg cgctcacgcg
     5221 cccgctctgg gattaagtga acaaagccca gaaggtcttg gttgctatca cgaaacccca
     5281 tccctctccc aataccggtc tcgaaaaagc tcgccgagcg actcatgtta aaggttacca
     5341 tgcattggta ctgattccat atattaacca tccggtccaa tttttcgtca ttggaactaa
     5401 cggaatacgt cgataacaaa tccgtccaat aatccttcaa agcggcgaaa gctgcgtcag
     5461 tctgttcgga cgtagcaaat cttgataata aagcgtgagc tctttcttta ttcacgacct
     5521 gttttgcgtc gtctgcccac ttctcctcat caggattttc cacgtaccca agcacataaa
     5581 ccaattctct ggattctcca ggcgctaaag acactgccac gctatgagag ccaataggat
     5641 accatccact tgcaacgcta tttgcacttt ctcccttaag gggaactgcg gcttcaccaa
     5701 gcgagttata ggctcccaca aatgtgtcgc gatctgtatc gaaaccctcg gcctgcgtat
     5761 taacggcgaa taccgcgtag tgatcgcgtc tctcccggta ctctgtacgg tgataaattg
     5821 cactgccatg cggagattct tgttccactt caacctctcc tatgctcaga ttacgctggt
     5881 agttagtttg atcgtcttgc gcgttccaaa gacaaaattc gacaaaagaa aacaacgtca
     5941 gggatttata cgagtcagag gtattagtga ccgtaacttt ctggacctcc gcgttctcac
     6001 cgactgggac gaaaaacaag gtctcaaccc gtaccccatt tctttcacct gtaatggtac
     6061 tgtagccaag accatgtctg gcttcaaaat gatctaagtc tgctttaaca ggaagccacg
     6121 atggcgtcca aacgtcccca ccgtcgttta cataaagata ccggcctccc gcgtctgctg
     6181 gtatgttgtt gtatctgtat cttgtcaatc tacgcatctt ggcatcgcga taaaacgaat
     6241 agccgccagc ctggtgcgac aaaagactga agaactgttc cgaacccaga tagttgatcc
     6301 aagggtacgg agtatgcggt gtggtgataa cgtactcgcg tgcttcatcg tcaaaatgac
     6361 catagcgggt cgacccgtga tggtgatggt gatggctgct gcccatatga tatctccttc
     6421 ttaaagttaa acaaaattat ttctagaggg gaattgttat ccgctcacaa ttcccctata
     6481 gtgagtcgta ttaatttcgc gggatcgact agtagatcaa aggatcttct tgagatcctt
     6541 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     6601 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     6661 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     6721 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     6781 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     6841 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     6901 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     6961 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     7021 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     7081 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     7141 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     7201 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     7261 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc
     7321 tccttacgca tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct
     7381 ctgatgccgc atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc
     7441 tgcgccccga cacccgccaa cg
//
