LOCUS       sequence-138074-        8451 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  IPTG inducible version of pZ8-1 plasmid, to which lacIq was added, 
            KanR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8451)
  AUTHORS   Cleto S, Jensen JV, Wendisch VF, Lu TK
  TITLE     Corynebacterium glutamicum Metabolic Engineering with CRISPR 
            Interference (CRISPRi).
  JOURNAL   ACS Synth Biol. 2016 Feb 16.
  PUBMED    26829286
REFERENCE   2  (bases 1 to 8451)
  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: "ACS Synth 
            Biol. 2016 Feb 16."
COMMENT     Sequence Label: pZ8-Ptac
FEATURES             Location/Qualifiers
     source          1..8451
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     268..290
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        368..445
                     /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             446..1528
                     /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
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(465..484)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(1645..1650,1651..1666,1667..1673)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1681..1697
                     /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     1686..1708
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(1780..1797)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1950..2036
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2128..2155
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2174..2265
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2484..2503)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             complement(6210..7025)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIADALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     6933..6952
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     7017..7036
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(7486..7503)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7620..8165)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(7700..7719)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cggtacgcct gcggccaaat ttgattcctt tcagctttgc ttcctgtcgg ccctcattcg
       61 tgcgctctag gatcagtgat ccgggcttat cgactgcacg gtgcaccaat gcttctggcg
      121 tcaggcagcc atcggaagct gtggtatggc cgctatcgct acgtgactgg gtcatggctg
      181 cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
      241 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
      301 catcaccgaa acgcgcgagg cagcagatca attcgcgcgc gaaggcgaag cggcatgcat
      361 ttacgttgac accatcgaat ggtgcaaaac ctttcgcggt atggcatgat agcgcccgga
      421 agagagtcaa ttcagggtgg tgaatgtgaa accagtaacg ttatacgatg tcgcagagta
      481 tgccggtgtc tcttatcaga ccgtttcccg cgtggtgaac caggccagcc acgtttctgc
      541 gaaaacgcgg gaaaaagtgg aagcggcgat ggcggagctg aattacattc ccaaccgcgt
      601 ggcacaacaa ctggcgggca aacagtcgtt gctgattggc gttgccacct ccagtctggc
      661 cctgcacgcg ccgtcgcaaa ttgtcgcggc gattaaatct cgcgccgatc aactgggtgc
      721 cagcgtggtg gtgtcgatgg tagaacgaag cggcgtcgaa gcctgtaaag cggcggtgca
      781 caatcttctc gcgcaacgcg tcagtgggct gatcattaac tatccgctgg atgaccagga
      841 tgccattgct gtggaagctg cctgcactaa tgttccggcg ttatttcttg atgtctctga
      901 ccagacaccc atcaacagta ttattttctc ccatgaagac ggtacgcgac tgggcgtgga
      961 gcatctggtc gcattgggtc accagcaaat cgcgctgtta gcgggcccat taagttctgt
     1021 ctcggcgcgt ctgcgtctgg ctggctggca taaatatctc actcgcaatc aaattcagcc
     1081 gatagcggaa cgggaaggcg actggagtgc catgtccggt tttcaacaaa ccatgcaaat
     1141 gctgaatgag ggcatcgttc ccactgcgat gctggttgcc aacgatcaga tggcgctggg
     1201 cgcaatgcgc gccattaccg agtccgggct gcgcgttggt gcggatatct cggtagtggg
     1261 atacgacgat accgaagaca gctcatgtta tatcccgccg tcaaccacca tcaaacagga
     1321 ttttcgcctg ctggggcaaa ccagcgtgga ccgcttgctg caactctctc agggccaggc
     1381 ggtgaagggc aatcagctgt tgcccgtctc actggtgaaa agaaaaacca ccctggcgcc
     1441 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
     1501 ggtttcccga ctggaaagcg ggcagtgact gtgcaggtcg taaatcactg cataattcgt
     1561 gtcgctcaag gcgcactccc gttctggata atgttttttg cgccgacatc ataacggttc
     1621 tggcaaatat tctgaaatga gctgttgaca attaatcatc ggctcgtata atgtgtggaa
     1681 ttgtgagcgg ataacaattt cacacaggaa acagaattcc cggggatccg tcgacctgca
     1741 gccaagcttc tgttttggcg gatgagagaa gattttcagc ctgatacaga ttaaatcaga
     1801 acgcagaagc ggtctgataa aacagaattt gcctggcggc agtagcgcgg tggtcccacc
     1861 tgaccccatg ccgaactcag aagtgaaacg ccgtagcgcc gatggtagtg tggggtctcc
     1921 ccatgcgaga gtagggaact gccaggcatc aaataaaacg aaaggctcag tcgaaagact
     1981 gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc
     2041 cgggagcgga tttgaacgtt gcgaagcaac ggcccggagg gtggcgggca ggacgcccgc
     2101 cataaactgc caggcatcaa attaagcaga aggccatcct gacggatggc ctttttgcgt
     2161 ttctacaaac tcttttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     2221 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     2281 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     2341 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     2401 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     2461 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg
     2521 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactgcggc
     2581 gtcgctgatc gccctggcga cgttgtgcgg gtggcttgtc cctgagggcg ctgcgacaga
     2641 tagctaaaaa tctgggtcag gatcgccgta gagcgcgcgt cgtcgattgg aggcttcccc
     2701 tttggttgac ggtcttcaat cgctctacgg cgatcctgac gcttttttgt tgcgtaccgt
     2761 cgatcgtttt atttctgtcg atcccgaaaa agtttttgcc ttttgtaaaa aacttctcgg
     2821 tcgccccgca aattttcgat tccagatttt ttaaaaacca agccagaaat acgacacacc
     2881 gtttgcagat aatctgtctt tcggaaaaat caagtgcgat acaaaatttt tagcacccct
     2941 gagctgcgca aagtcccgct tcgtgaaaat tttcgtgccg cgtgattttc cgccaaaaac
     3001 tttaacgaac gttcgttata atggtgtcat gaccttcacg acgaagtact aaaattggcc
     3061 cgaatcatca gctatggatc tctctgatgt cgcgctggag tccgacgcgc tcgatgctgc
     3121 cgtcgattta aaaacggtga tcggattttt ccgagctctc gatacgacgg acgcgccagc
     3181 atcacgagac tgggccagtg ccgcgagcga cctagaaact ctcgtggcgg atcttgagga
     3241 gctggctgac gagctgcgtg ctcggccagc gccaggagga cgcacagtag tggaggatgc
     3301 aatcagttgc gcctactgcg gtggcctgat tcctccccgg cctgacccgc gaggacggcg
     3361 cgcaaaatat tgctcagatg cgtgtcgtgc cgcagccagc cgcgagcgcg ccaacaaacg
     3421 ccacgccgag gagctggagg cggctaggtc gcaaatggcg ctggaagtgc gtcccccgag
     3481 cgaaattttg gccatggtcg tcacagagct ggaagcggca gcgagaatta tccgcgatcg
     3541 tggcgcggtg cccgcaggca tgacaaacat cgtaaatgcc gcgtttcgtg tggccgtggc
     3601 cgcccaggac gtgtcagcgc cgccaccacc tgcaccgaat cggcagcagc gtcgcgcgtc
     3661 gaaaaagcgc acaggcggca agaagcgata agctgcacga atacctgaaa aatgttgaac
     3721 gccccgtgag cggtaactca cagggcgtcg gctaaccccc agtccaaacc tgggagaaag
     3781 cgctcaaaaa tgactctagc ggattcacga gacattgaca caccggcctg gaaattttcc
     3841 gctgatctgt tcgacaccca tcccgagctc gcgctgcgat cacgtggctg gacgagcgaa
     3901 gaccgccgca aattcctcgc tcacctgggc agagaaaatt tccagggcag caagacccgc
     3961 gacttcgcca gcgcttggat caaagacccg gacacgggag aaacacagcc gaagttatac
     4021 cgagttggtt caaaatcgct tgcccggtgc cagtatgttg ctctgacgca cgcgcagcac
     4081 gcagccgtgc ttgtcctgga cattgatgtg ccgagccacc aggccggcgg gaaaatcgag
     4141 cacgtaaacc ccgaggtcta cgcgattttg gagcgctggg cacgcctgga aaaagcgcca
     4201 gcttggatcg gcgtgaatcc actgagcggg aaatgccagc tcatctggct cattgatccg
     4261 gtgtatgccg cagcaggcat gagcagcccg aatatgcgcc tgctggctgc aacgaccgag
     4321 gaaatgaccc gcgttttcgg cgctgaccag gctttttcac ataggctgag ccgtggccac
     4381 tgcactctcc gacgatccca gccgtaccgc tggcatgccc agcacaatcg cgtggatcgc
     4441 ctagctgatc ttatggaggt tgctcgcatg atctcaggca cagaaaaacc taaaaaacgc
     4501 tatgagcagg agttttctag cggacgggca cgtatcgaag cggcaagaaa agccactgcg
     4561 gaagcaaaag cacttgccac gcttgaagca agcctgccga gcgccgctga agcgtctgga
     4621 gagctgatcg acggcgtccg tgtcctctgg actgctccag ggcgtgccgc ccgtgatgag
     4681 acggcttttc gccacgcttt gactgtggga taccagttaa aagcggctgg tgagcgccta
     4741 aaagacacca agggtcatcg agcctacgag cgtgcctaca ccgtcgctca ggcggtcgga
     4801 ggaggccgtg agcctgatct gccgccggac tgtgaccgcc agacggattg gccgcgacgt
     4861 gtgcgcggct acgtcgctaa aggccagcca gtcgtccctg ctcgtcagac agagacgcag
     4921 agccagccga ggcgaaaagc tctggccact atgggaagac gtggcggtaa aaaggccgca
     4981 gaacgctgga aagacccaaa cagtgagtac gcccgagcac agcgagaaaa actagctaag
     5041 tccagtcaac gacaagctag gaaagctaaa ggaaatcgct tgaccattgc aggttggttt
     5101 atgactgttg agggagagac tggctcgtgg ccgacaatca atgaagctat gtctgaattt
     5161 agcgtgtcac gtcagaccgt gaatagagca cttaaggtct gcgggcattg aacttccacg
     5221 aggacgccga aagcttccca gtaaatgtgc catctcgtag gcagaaaacg gttcccccgt
     5281 agggtctctc tcttggcctc ctttctaggt cgggctgatt gctcttgaag ctctctaggg
     5341 gggctcacac cataggcaga taacgttccc caccggctcg cctcgtaagc gcacaaggac
     5401 tgctcccaaa gatcttcaaa gccactgccg cgactgcctt cgcgaagcct tgccccgcgg
     5461 aaatttcctc caccgagttc gtgcacaccc ctatgccaag cttctttcac cctaaattcg
     5521 agagattgga ttcttaccgt ggaaattctt cgcaaaaatc gtcccctgat cgcccttgcg
     5581 acgttggcgt cggtgccgct ggttgcgctt ggcttgaccg acttgatcct ccggcgttca
     5641 gcctgtgcca cagccgacag gatggtgacc accatttgcc ccatatcacc gtcggtactg
     5701 atcccgtcgt caataaaccg aaccgctaca ccctgagcat caaactcttt tatcagttgg
     5761 atcatgtcgg cgtgtcgcgg ccaagacggt cgagcttctt caccagaatg acatcacctt
     5821 cctccacctt catcctcagc aaatccagcc cttcccgatc tgttgaactg ccggatgcct
     5881 tgtcggtaaa gatgcggtta gcttttaccc ctgcatcttt gagcgctgag gtctgcctcg
     5941 tgaagaaggt gttgctgact cataccaggc ctgaatcgcc ccatcatcca gccagaaagt
     6001 gagggagcca cggttgatga gagctttgtt gtaggtggac cagttggtga ttttgaactt
     6061 ttgctttgcc acggaacggt ctgcgttgtc gggaagatgc gtgatctgat ccttcaactc
     6121 agcaaaagtt cgatttattc aacaaagccg ccgtcccgtc aagtcagcgt aatgctctgc
     6181 cagtgttaca accaattaac caattctgat tagaaaaact catcgagcat caaatgaaac
     6241 tgcaatttat tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat
     6301 gaaggagaaa actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg
     6361 attccgactc gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta
     6421 tcaagtgaga aatcaccatg agtgacgact gaatccggtg agaatggcaa aagcttatgc
     6481 atttctttcc agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca
     6541 tcaaccaaac cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg
     6601 ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca
     6661 tcagcaatat tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg
     6721 gggatcgcag tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc
     6781 ggaagaggca taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg
     6841 gcaacgctac ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat
     6901 cgatagattg tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa
     6961 tcagcatcca tgttggaatt taatcgcggc ctcgagcaag acgtttcccg ttgaatatgg
     7021 ctcataacac cccttgtatt actgtttatg taagcagaca gttttattgt tcatgatgat
     7081 atatttttat cttgtgcaat gtaacatcag agattttgag acacaacgtg gctttgttga
     7141 ataaatcgaa cttttgctga gttgaaggat cagatcacgc atcttcccga caacgcagac
     7201 cgttccgtgg caaagcaaaa gttcaaaatc accaactggt ccacctacaa caaagctctc
     7261 atcaaccgtg gctccctcac tttctggctg gatgatgggg cgattcaggc ctggtatgag
     7321 tcagcaacac cttcttcacg aggcagacct cagcgctagc ggagtgtata ctggcttact
     7381 atgttggcac tgatgagggt gtcagtgaag tgcttcatgt ggcaggagaa aaaaggctgc
     7441 accggtgcgt cagcagaata tgtgatacag gatatattcc gcttcctcgc tcactgactc
     7501 gctacgctcg gtcgttcgac tgcggcgagc ggaaatggct tacgaacggg gcggagattt
     7561 cctggaagat gccaggaaga tacttaacag ggaagtgaga gggccgcggc aaagccgttt
     7621 ttccataggc tccgcccccc tgacaagcat cacgaaatct gacgctcaaa tcagtggtgg
     7681 cgaaacccga caggactata aagataccag gcgtttcccc ctggcggctc cctcgtgcgc
     7741 tctcctgttc ctgcctttcg gtttaccggt gtcattccgc tgttatggcc gcgtttgtct
     7801 cattccacgc ctgacactca gttccgggta ggcagttcgc tccaagctgg actgtatgca
     7861 cgaacccccc gttcagtccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     7921 cccggaaaga catgcaaaag caccactggc agcagccact ggtaattgat ttagaggagt
     7981 tagtcttgaa gtcatgcgcc ggttaaggct aaactgaaag gacaagtttt ggtgactgcg
     8041 ctcctccaag ccagttacct cggttcaaag agttggtagc tcagagaacc ttcgaaaaac
     8101 cgccctgcaa ggcggttttt tcgttttcag agcaagagat tacgcgcaga ccaaaacgat
     8161 ctcaagaaga tcatcttatt aaggggtctg acgctcagtg gaacgaaaac tcacgttaag
     8221 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     8281 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     8341 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     8401 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc a
//
