LOCUS       sequence-360587-        3952 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression vector, low to medium copy number, tac promoter lacI, 
            kanamycin resistance, p15A origin of replication.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3952)
  AUTHORS   Stratmann T, Madhusudan S, Schnetz K
  TITLE     Regulation of the yjjQ-bglJ operon, encoding LuxR-type transcription
            factors, and the divergent yjjP gene by H-NS and LeuO.
  JOURNAL   J Bacteriol. 2008 Feb;190(3):926-35. Epub 2007 Nov 30.
  PUBMED    18055596
REFERENCE   2  (bases 1 to 3952)
  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: "J 
            Bacteriol."; date: "2008-02"; volume: "190(3)"; pages: "926-35. Epub
            2007 Nov 30"
COMMENT     Sequence Label: pKESK22
FEATURES             Location/Qualifiers
     source          1..3952
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             78..1160
                     /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     complement(97..116)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(1209..1214,1215..1230,1231..1237)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1245..1261
                     /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     1250..1272
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             complement(1888..2703)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(1888..2703)
                     /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
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2611..2630
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2695..2714
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(3164..3181)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3298..3843)
                     /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(3378..3397)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(3952,1..77)
                     /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 acaccatcga atggtgcaaa acctttcgcg gtatggcatg atagcgcccg gaagagagtc
       61 aattcagggt ggtgaatgtg aaaccagtaa cgttatacga tgtcgcagag tatgccggtg
      121 tctcttatca gaccgtttcc cgcgtggtga accaggccag ccacgtttct gcgaaaacgc
      181 gggaaaaagt ggaagcggcg atggcggagc tgaattacat tcccaaccgc gtggcacaac
      241 aactggcggg caaacagtcg ttgctgattg gcgttgccac ctccagtctg gccctgcacg
      301 cgccgtcgca aattgtcgcg gcgattaaat ctcgcgccga tcaactgggt gccagcgtgg
      361 tggtgtcgat ggtagaacga agcggcgtcg aagcctgtaa agcggcggtg cacaatcttc
      421 tcgcgcaacg cgtcagtggg ctgatcatta actatccgct ggatgaccag gatgccattg
      481 ctgtggaagc tgcctgcact aatgttccgg cgttatttct tgatgtctct gaccagacac
      541 ccatcaacag tattattttc tcccatgaag acggtacgcg actgggcgtg gagcatctgg
      601 tcgcattggg tcaccagcaa atcgcgctgt tagcgggccc attaagttct gtctcggcgc
      661 gtctgcgtct ggctggctgg cataaatatc tcactcgcaa tcaaattcag ccgatagcgg
      721 aacgggaagg cgactggagt gccatgtccg gttttcaaca aaccatgcaa atgctgaatg
      781 agggcatcgt tcccactgcg atgctggttg ccaacgatca gatggcgctg ggcgcaatgc
      841 gcgccattac cgagtccggg ctgcgcgttg gtgcggatat ctcggtagtg ggatacgacg
      901 ataccgaaga cagctcatgt tatatcccgc cgttaaccac catcaaacag gattttcgcc
      961 tgctggggca aaccagcgtg gaccgcttgc tgcaactctc tcagggccag gcggtgaagg
     1021 gcaatcagct gttgcccgtc tcactggtga aaagaaaaac caccctggcg cccaatacgc
     1081 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     1141 gactggaaag cgggcagtga gcgcaacgca attaatgtaa gttagctcaa atattctgaa
     1201 atgagctgtt gacaattaat catcggctcg tataatgtgt ggaattgtga gcggataaca
     1261 atttcacaca ggaaacagaa ttcgctagcc cgggtctaga ggatcctccg gcgttcagcc
     1321 tgtgccacag ccgacaggat ggtgaccacc atttgcccca tatcaccgtc ggtactgatc
     1381 ccgtcgtcaa taaaccgaac cgctacaccc tgagcatcaa actcttttat cagttggatc
     1441 atgtcggcgg tgtcgcggcc aagacggtcg agcttcttca ccagaatgac atcaccttcc
     1501 tccaccttca tcctcagcaa atccagccct tcccgatctg ttgaactgcc ggatgccttg
     1561 tcggtaaaga tgcggttagc ttttacccct gcatctttga gcgctgaggt ctgcctcgtg
     1621 aagaaggtgt tgctgactca taccaggcct gaatcgcccc atcatccagc cagaaagtga
     1681 gggagccacg gttgatgaga gctttgttgt aggtggacca gttggtgatt ttgaactttt
     1741 gctttgccac ggaacggtct gcgttgtcgg gaagatgcgt gatctgatcc ttcaactcag
     1801 caaaagttcg atttattcaa caaagccgcc gtcccgtcaa gtcagcgtaa tgctctgcca
     1861 gtgttacaac caattaacca attctgatta gaaaaactca tcgagcatca aatgaaactg
     1921 caatttattc atatcaggat tatcaatacc atatttttga aaaagccgtt tctgtaatga
     1981 aggagaaaac tcaccgaggc agttccatag gatggcaaga tcctggtatc ggtctgcgat
     2041 tccgactcgt ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa taaggttatc
     2101 aagtgagaaa tcaccatgag tgacgactga atccggtgag aatggcaaaa gcttatgcat
     2161 ttctttccag acttgttcaa caggccagcc attacgctcg tcatcaaaat cactcgcatc
     2221 aaccaaaccg ttattcattc gtgattgcgc ctgagcgaga cgaaatacgc gatcgctgtt
     2281 aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc aggaacactg ccagcgcatc
     2341 aacaatattt tcacctgaat caggatattc ttctaatacc tggaatgctg ttttcccggg
     2401 gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg ataaaatgct tgatggtcgg
     2461 aagaggcata aattccgtca gccagtttag tctgaccatc tcatctgtaa catcattggc
     2521 aacgctacct ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc catacaatcg
     2581 atagattgtc gcacctgatt gcccgacatt atcgcgagcc catttatacc catataaatc
     2641 agcatccatg ttggaattta atcgcggcct cgagcaagac gtttcccgtt gaatatggct
     2701 cataacaccc cttgtattac tgtttatgta agcagacagt tttattgttc atgatgatat
     2761 atttttatct tgtgcaatgt aacatcagag attttgagac acaacgtggc tttgttgaat
     2821 aaatcgaact tttgctgagt tgaaggatca gatcacgcat cttcccgaca acgcagaccg
     2881 ttccgtggca aagcaaaagt tcaaaatcac caactggtcc acctacaaca aagctctcat
     2941 caaccgtggc tccctcactt tctggctgga tgatggggcg attcaggcct ggtatgagtc
     3001 agcaacacct tcttcacgag gcagacctca gcgctagcgg agtgtatact ggcttactat
     3061 gttggcactg atgagggtgt cagtgaagtg cttcatgtgg caggagaaaa aaggctgcac
     3121 cggtgcgtca gcagaatatg tgatacagga tatattccgc ttcctcgctc actgactcgc
     3181 tacgctcggt cgttcgactg cggcgagcgg aaatggctta cgaacggggc ggagatttcc
     3241 tggaagatgc caggaagata cttaacaggg aagtgagagg gccgcggcaa agccgttttt
     3301 ccataggctc cgcccccctg acaagcatca cgaaatctga cgctcaaatc agtggtggcg
     3361 aaacccgaca ggactataaa gataccaggc gtttccccct ggcggctccc tcgtgcgctc
     3421 tcctgttcct gcctttcggt ttaccggtgt cattccgctg ttatggccgc gtttgtctca
     3481 ttccacgcct gacactcagt tccgggtagg cagttcgctc caagctggac tgtatgcacg
     3541 aaccccccgt tcagtccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     3601 cggaaagaca tgcaaaagca ccactggcag cagccactgg taattgattt agaggagtta
     3661 gtcttgaagt catgcgccgg ttaaggctaa actgaaagga caagttttgg tgactgcgct
     3721 cctccaagcc agttacctcg gttcaaagag ttggtagctc agagaacctt cgaaaaaccg
     3781 ccctgcaagg cggttttttc gttttcagag caagagatta cgcgcagacc aaaacgatct
     3841 caagaagatc atcttattaa ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     3901 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttggt cg
//
