LOCUS       sequence-138071-        7268 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pZ8-1 with the PprpD2 propionate-inducible promoter instead of Ptac,
            KanR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7268)
  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 7268)
  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-Prp
FEATURES             Location/Qualifiers
     source          1..7268
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(597..614)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      767..853
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      945..972
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        991..1082
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1301..1320)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             complement(5027..5842)
                     /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     5750..5769
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5834..5853
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(6303..6320)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6437..6982)
                     /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(6517..6536)
                     /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 tgtgcaggtc gtaaatcact gcataattcg
      181 tgtcgctcaa ggcgcactcc cgttctggat aatgtttttt gcgccgacat cataacggtt
      241 ctggcaaata ttctgaaatg agctgctcca gcgtccaaga atatgccccc gcgcgccggg
      301 tggggagcga agggaacccc caaggaattg gcgttgaggt ggtgattttg catgttttac
      361 tcaaaatcac tttgatggtc acaaaattac acaactttta cagtgaccta cattgctttt
      421 taaagaatta gtgtggtgtg cgatccccgg gtacccagct tgagtaggac aaatccgccg
      481 agcttcgacg agattttcag gagctaagga agctaaacac acaggaaaca gaattcccgg
      541 ggatccgtcg acctgcagcc aagcttctgt tttggcggat gagagaagat tttcagcctg
      601 atacagatta aatcagaacg cagaagcggt ctgataaaac agaatttgcc tggcggcagt
      661 agcgcggtgg tcccacctga ccccatgccg aactcagaag tgaaacgccg tagcgccgat
      721 ggtagtgtgg ggtctcccca tgcgagagta gggaactgcc aggcatcaaa taaaacgaaa
      781 ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcct
      841 gagtaggaca aatccgccgg gagcggattt gaacgttgcg aagcaacggc ccggagggtg
      901 gcgggcagga cgcccgccat aaactgccag gcatcaaatt aagcagaagg ccatcctgac
      961 ggatggcctt tttgcgtttc tacaaactct tttgtttatt tttctaaata cattcaaata
     1021 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     1081 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     1141 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     1201 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     1261 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     1321 cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     1381 tggttgagta ctgcggcgtc gctgatcgcc ctggcgacgt tgtgcgggtg gcttgtccct
     1441 gagggcgctg cgacagatag ctaaaaatct gggtcaggat cgccgtagag cgcgcgtcgt
     1501 cgattggagg cttccccttt ggttgacggt cttcaatcgc tctacggcga tcctgacgct
     1561 tttttgttgc gtaccgtcga tcgttttatt tctgtcgatc ccgaaaaagt ttttgccttt
     1621 tgtaaaaaac ttctcggtcg ccccgcaaat tttcgattcc agatttttta aaaaccaagc
     1681 cagaaatacg acacaccgtt tgcagataat ctgtctttcg gaaaaatcaa gtgcgataca
     1741 aaatttttag cacccctgag ctgcgcaaag tcccgcttcg tgaaaatttt cgtgccgcgt
     1801 gattttccgc caaaaacttt aacgaacgtt cgttataatg gtgtcatgac cttcacgacg
     1861 aagtactaaa attggcccga atcatcagct atggatctct ctgatgtcgc gctggagtcc
     1921 gacgcgctcg atgctgccgt cgatttaaaa acggtgatcg gatttttccg agctctcgat
     1981 acgacggacg cgccagcatc acgagactgg gccagtgccg cgagcgacct agaaactctc
     2041 gtggcggatc ttgaggagct ggctgacgag ctgcgtgctc ggccagcgcc aggaggacgc
     2101 acagtagtgg aggatgcaat cagttgcgcc tactgcggtg gcctgattcc tccccggcct
     2161 gacccgcgag gacggcgcgc aaaatattgc tcagatgcgt gtcgtgccgc agccagccgc
     2221 gagcgcgcca acaaacgcca cgccgaggag ctggaggcgg ctaggtcgca aatggcgctg
     2281 gaagtgcgtc ccccgagcga aattttggcc atggtcgtca cagagctgga agcggcagcg
     2341 agaattatcc gcgatcgtgg cgcggtgccc gcaggcatga caaacatcgt aaatgccgcg
     2401 tttcgtgtgg ccgtggccgc ccaggacgtg tcagcgccgc caccacctgc accgaatcgg
     2461 cagcagcgtc gcgcgtcgaa aaagcgcaca ggcggcaaga agcgataagc tgcacgaata
     2521 cctgaaaaat gttgaacgcc ccgtgagcgg taactcacag ggcgtcggct aacccccagt
     2581 ccaaacctgg gagaaagcgc tcaaaaatga ctctagcgga ttcacgagac attgacacac
     2641 cggcctggaa attttccgct gatctgttcg acacccatcc cgagctcgcg ctgcgatcac
     2701 gtggctggac gagcgaagac cgccgcaaat tcctcgctca cctgggcaga gaaaatttcc
     2761 agggcagcaa gacccgcgac ttcgccagcg cttggatcaa agacccggac acgggagaaa
     2821 cacagccgaa gttataccga gttggttcaa aatcgcttgc ccggtgccag tatgttgctc
     2881 tgacgcacgc gcagcacgca gccgtgcttg tcctggacat tgatgtgccg agccaccagg
     2941 ccggcgggaa aatcgagcac gtaaaccccg aggtctacgc gattttggag cgctgggcac
     3001 gcctggaaaa agcgccagct tggatcggcg tgaatccact gagcgggaaa tgccagctca
     3061 tctggctcat tgatccggtg tatgccgcag caggcatgag cagcccgaat atgcgcctgc
     3121 tggctgcaac gaccgaggaa atgacccgcg ttttcggcgc tgaccaggct ttttcacata
     3181 ggctgagccg tggccactgc actctccgac gatcccagcc gtaccgctgg catgcccagc
     3241 acaatcgcgt ggatcgccta gctgatctta tggaggttgc tcgcatgatc tcaggcacag
     3301 aaaaacctaa aaaacgctat gagcaggagt tttctagcgg acgggcacgt atcgaagcgg
     3361 caagaaaagc cactgcggaa gcaaaagcac ttgccacgct tgaagcaagc ctgccgagcg
     3421 ccgctgaagc gtctggagag ctgatcgacg gcgtccgtgt cctctggact gctccagggc
     3481 gtgccgcccg tgatgagacg gcttttcgcc acgctttgac tgtgggatac cagttaaaag
     3541 cggctggtga gcgcctaaaa gacaccaagg gtcatcgagc ctacgagcgt gcctacaccg
     3601 tcgctcaggc ggtcggagga ggccgtgagc ctgatctgcc gccggactgt gaccgccaga
     3661 cggattggcc gcgacgtgtg cgcggctacg tcgctaaagg ccagccagtc gtccctgctc
     3721 gtcagacaga gacgcagagc cagccgaggc gaaaagctct ggccactatg ggaagacgtg
     3781 gcggtaaaaa ggccgcagaa cgctggaaag acccaaacag tgagtacgcc cgagcacagc
     3841 gagaaaaact agctaagtcc agtcaacgac aagctaggaa agctaaagga aatcgcttga
     3901 ccattgcagg ttggtttatg actgttgagg gagagactgg ctcgtggccg acaatcaatg
     3961 aagctatgtc tgaatttagc gtgtcacgtc agaccgtgaa tagagcactt aaggtctgcg
     4021 ggcattgaac ttccacgagg acgccgaaag cttcccagta aatgtgccat ctcgtaggca
     4081 gaaaacggtt cccccgtagg gtctctctct tggcctcctt tctaggtcgg gctgattgct
     4141 cttgaagctc tctagggggg ctcacaccat aggcagataa cgttccccac cggctcgcct
     4201 cgtaagcgca caaggactgc tcccaaagat cttcaaagcc actgccgcga ctgccttcgc
     4261 gaagccttgc cccgcggaaa tttcctccac cgagttcgtg cacaccccta tgccaagctt
     4321 ctttcaccct aaattcgaga gattggattc ttaccgtgga aattcttcgc aaaaatcgtc
     4381 ccctgatcgc ccttgcgacg ttggcgtcgg tgccgctggt tgcgcttggc ttgaccgact
     4441 tgatcctccg gcgttcagcc tgtgccacag ccgacaggat ggtgaccacc atttgcccca
     4501 tatcaccgtc ggtactgatc ccgtcgtcaa taaaccgaac cgctacaccc tgagcatcaa
     4561 actcttttat cagttggatc atgtcggcgt gtcgcggcca agacggtcga gcttcttcac
     4621 cagaatgaca tcaccttcct ccaccttcat cctcagcaaa tccagccctt cccgatctgt
     4681 tgaactgccg gatgccttgt cggtaaagat gcggttagct tttacccctg catctttgag
     4741 cgctgaggtc tgcctcgtga agaaggtgtt gctgactcat accaggcctg aatcgcccca
     4801 tcatccagcc agaaagtgag ggagccacgg ttgatgagag ctttgttgta ggtggaccag
     4861 ttggtgattt tgaacttttg ctttgccacg gaacggtctg cgttgtcggg aagatgcgtg
     4921 atctgatcct tcaactcagc aaaagttcga tttattcaac aaagccgccg tcccgtcaag
     4981 tcagcgtaat gctctgccag tgttacaacc aattaaccaa ttctgattag aaaaactcat
     5041 cgagcatcaa atgaaactgc aatttattca tatcaggatt atcaatacca tatttttgaa
     5101 aaagccgttt ctgtaatgaa ggagaaaact caccgaggca gttccatagg atggcaagat
     5161 cctggtatcg gtctgcgatt ccgactcgtc caacatcaat acaacctatt aatttcccct
     5221 cgtcaaaaat aaggttatca agtgagaaat caccatgagt gacgactgaa tccggtgaga
     5281 atggcaaaag cttatgcatt tctttccaga cttgttcaac aggccagcca ttacgctcgt
     5341 catcaaaatc actcgcatca accaaaccgt tattcattcg tgattgcgcc tgagcgagac
     5401 gaaatacgcg atcgctgtta aaaggacaat tacaaacagg aatcgaatgc aaccggcgca
     5461 ggaacactgc cagcgcatca gcaatatttt cacctgaatc aggatattct tctaatacct
     5521 ggaatgctgt tttcccgggg atcgcagtgg tgagtaacca tgcatcatca ggagtacgga
     5581 taaaatgctt gatggtcgga agaggcataa attccgtcag ccagtttagt ctgaccatct
     5641 catctgtaac atcattggca acgctacctt tgccatgttt cagaaacaac tctggcgcat
     5701 cgggcttccc atacaatcga tagattgtcg cacctgattg cccgacatta tcgcgagccc
     5761 atttataccc atataaatca gcatccatgt tggaatttaa tcgcggcctc gagcaagacg
     5821 tttcccgttg aatatggctc ataacacccc ttgtattact gtttatgtaa gcagacagtt
     5881 ttattgttca tgatgatata tttttatctt gtgcaatgta acatcagaga ttttgagaca
     5941 caacgtggct ttgttgaata aatcgaactt ttgctgagtt gaaggatcag atcacgcatc
     6001 ttcccgacaa cgcagaccgt tccgtggcaa agcaaaagtt caaaatcacc aactggtcca
     6061 cctacaacaa agctctcatc aaccgtggct ccctcacttt ctggctggat gatggggcga
     6121 ttcaggcctg gtatgagtca gcaacacctt cttcacgagg cagacctcag cgctagcgga
     6181 gtgtatactg gcttactatg ttggcactga tgagggtgtc agtgaagtgc ttcatgtggc
     6241 aggagaaaaa aggctgcacc ggtgcgtcag cagaatatgt gatacaggat atattccgct
     6301 tcctcgctca ctgactcgct acgctcggtc gttcgactgc ggcgagcgga aatggcttac
     6361 gaacggggcg gagatttcct ggaagatgcc aggaagatac ttaacaggga agtgagaggg
     6421 ccgcggcaaa gccgtttttc cataggctcc gcccccctga caagcatcac gaaatctgac
     6481 gctcaaatca gtggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     6541 gcggctccct cgtgcgctct cctgttcctg cctttcggtt taccggtgtc attccgctgt
     6601 tatggccgcg tttgtctcat tccacgcctg acactcagtt ccgggtaggc agttcgctcc
     6661 aagctggact gtatgcacga accccccgtt cagtccgacc gctgcgcctt atccggtaac
     6721 tatcgtcttg agtccaaccc ggaaagacat gcaaaagcac cactggcagc agccactggt
     6781 aattgattta gaggagttag tcttgaagtc atgcgccggt taaggctaaa ctgaaaggac
     6841 aagttttggt gactgcgctc ctccaagcca gttacctcgg ttcaaagagt tggtagctca
     6901 gagaaccttc gaaaaaccgc cctgcaaggc ggttttttcg ttttcagagc aagagattac
     6961 gcgcagacca aaacgatctc aagaagatca tcttattaag gggtctgacg ctcagtggaa
     7021 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     7081 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     7141 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     7201 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     7261 tggcccca
//
