LOCUS       Exported                4571 bp DNA     circular SYN 21-JUL-2025
DEFINITION  Cloning vector pKK233-2, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4571)
  AUTHORS   Amann E, Brosius J.
  TITLE     'ATG vectors' for regulated high-level expression of cloned genes in
            Escherichia coli
  JOURNAL   Gene 40 (2-3), 183-190 (1985)
  PUBMED    3007288
REFERENCE   2  (bases 1 to 4571)
  AUTHORS   Kitts PA.
  TITLE     CLONTECH Vectors On Disc version 1.3
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 4571)
  AUTHORS   Kitts PA.
  TITLE     Direct Submission
  JOURNAL   Submitted (07-OCT-1993) Paul A. Kitts, CLONTECH Laboratories, Inc., 
            1020 East Meadow Circle, Palo Alto, CA 94303, USA
REFERENCE   4  (bases 1 to 4571)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 4571)
  TITLE     Direct Submission
REFERENCE   6  (bases 1 to 4571)
  TITLE     Direct Submission
REFERENCE   7  (bases 1 to 4571)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Gene 40
            (2-3), 183-190 (1985)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Submitted
            (07-OCT-1993) Paul A. Kitts, CLONTECH Laboratories, Inc., 1020 East
            Meadow Circle, Palo Alto, CA 94303, USA"
COMMENT     SGRef: number: 4; type: "Journal Article"
COMMENT     SGRef: number: 5; type: "Journal Article"
COMMENT     SGRef: number: 6; type: "Journal Article"
COMMENT     This vector can be obtained from CLONTECH Laboratories, Inc., 1020 
            East Meadow Circle, Palo Alto, CA 94303, USA. To place an order call
            (415) 424-8222 or (800) 662-2566, extension 1. International 
            customers, please contact your local distributor.  For technical 
            information, call (415) 424- 8222 or (800) 662-2566, extension 3. 
            This sequence was compiled by Jurgen Brosius. If you suspect there 
            is an error in this sequence, please contact CLONTECH's Technical 
            Service Department at (415) 424-8222 or (800) 662-2566, extension 3 
            or E-mail TECH@CLONTECH.COM.
FEATURES             Location/Qualifiers
     source          1..4571
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        184..212
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    220..236
                     /label=lac operator
                     /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)."
     terminator      477..563
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      655..682
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     promoter        702..792
                     /label=AmpR promoter
     CDS             793..1650
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1824..2412
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(2598..2738)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(2840..3031)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3670..4455)
                     /codon_start=1
                     /label=tetracycline efflux MFS transporter Tet(C)
                     /translation="MSACFGVGMVAGPVAGGLLGAISLHAPFLAAAVLNGLNLLLGCFL
                     MQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIMQLVGQVPAALWVIFGED
                     RFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAIIAGMAADALGYVLLAFAT
                     RGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSLAALTSLTSIIGPLIVTA
                     IYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
ORIGIN
        1 gggcttatcg actgcacggt gcaccaatgc ttctggcgtc aggcagccat cggaagctgt
       61 ggtatggctg tgcaggtcgt aaatcactgc ataattcgtg tcgctcaagg cgcactcccg
      121 ttctggataa tgttttttgc gccgacatca taacggttct ggcaaatatt ctgaaatgag
      181 ctgttgacaa ttaatcatcg gctcgtataa tgtgtggaat tgtgagcgga taacaatttc
      241 acacaggaaa cagaccatgg ctgcagccaa gcttggctgt tttggcggat gagagaagat
      301 tttcagcctg atacagatta aatcagaacg cagaagcggt ctgataaaac agaatttgcc
      361 tggcggcagt agcgcggtgg tcccacctga ccccatgccg aactcagaag tgaaacgccg
      421 tagcgccgat ggtagtgtgg ggtctcccca tgcgagagta gggaactgcc aggcatcaaa
      481 taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga
      541 acgctctcct gagtaggaca aatccgccgg gagcggattt gaacgttgcg aagcaacggc
      601 ccggagggtg gcgggcagga cgcccgccat aaactgccag gcatcaaatt aagcagaagg
      661 ccatcctgac ggatggcctt tttgcgtttc tacaaactct tttgtttatt tttctaaata
      721 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
      781 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
      841 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
      901 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
      961 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     1021 gcggtattat cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     1081 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     1141 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     1201 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     1261 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     1321 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     1381 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     1441 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     1501 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     1561 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     1621 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     1681 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     1741 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     1801 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     1861 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     1921 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
     1981 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     2041 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     2101 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     2161 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     2221 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     2281 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     2341 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     2401 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     2461 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     2521 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     2581 gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     2641 caccgcatat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
     2701 acactccgct atcgctacgt gactgggtca tggctgcgcc ccgacacccg ccaacacccg
     2761 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     2821 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgaggcagc
     2881 tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg
     2941 cgtccagctc gttgagtttc tccagaagcg ttaatgtctg gcttctgata aagcgggcca
     3001 tgttaagggc ggttttttcc tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt
     3061 tcatgggggt aatgataccg atgaaacgag agaggatgct cacgatacgg gttactgatg
     3121 atgaacatgc ccggttactg gaacgttgtg agggtaaaca actggcggta tggatgcggc
     3181 gggaccagag aaaaatcact cagggtcaat gccagcgctt cgttaataca gatgtaggtg
     3241 ttccacaggg tagccagcag catcctgcga tgcagatccg gaacataatg gtgcagggcg
     3301 ctgacttccg cgtttccaga ctttacgaaa cacggaaacc gaagaccatt catgttgttg
     3361 ctcaggtcgc agacgttttg cagcagcagt cgcttcacgt tcgctcgcgt atcggtgatt
     3421 cattctgcta accagtaagg caaccccgcc agcctagccg ggtcctcaac gacaggagca
     3481 cgatcatgcg cacccgtggc caggacccaa cgctgcccga gatgcgccgc gtgcggctgc
     3541 tggagatggc ggacgcgatg gatatgttct gccaagggtt ggtttgcgca ttcacagttc
     3601 tccgcaagaa ttgattggct ccaattcttg gagtggtgaa tccgttagcg aggtgccgcc
     3661 ggcttccatt caggtcgagg tggcccggct ccatgcaccg cgacgcaacg cggggaggca
     3721 gacaaggtat agggcggcgc ctacaatcca tgccaacccg ttccatgtgc tcgccgaggc
     3781 ggcataaatc gccgtgacga tcagcggtcc aatgatcgaa gttaggctgg taagagccgc
     3841 gagcgatcct tgaagctgtc cctgatggtc gtcatctacc tgcctggaca gcatggcctg
     3901 caacgcgggc atcccgatgc cgccggaagc gagaagaatc ataatgggga aggccatcca
     3961 gcctcgcgtc gcgaacgcca gcaagacgta gcccagcgcg tcggccgcca tgccggcgat
     4021 aatggcctgc ttctcgccga aacgtttggt ggcgggacca gtgacgaagg cttgagcgag
     4081 ggcgtgcaag attccgaata ccgcaagcga caggccgatc atcgtcgcgc tccagcgaaa
     4141 gcggtcctcg ccgaaaatga cccagagcgc tgccggcacc tgtcctacga gttgcatgat
     4201 aaagaagaca gtcataagtg cggcgacgat agtcatgccc cgcgcccacc ggaaggagct
     4261 gactgggttg aaggctctca agggcatcgg tcgacgctct cccttatgcg actcctgcat
     4321 taggaagcag cccagtagta ggttgaggcc gttgagcacc gccgccgcaa ggaatggtgc
     4381 atgcaaggag atggcgccca acagtccccc ggccacgggg cctgccacca tacccacgcc
     4441 gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct tccccatcgg tgatgtcggc
     4501 gatataggcg ccagcaaccg cacctgtggc gccggtgatg ccggccacga tgcgtccggc
     4561 gtagaggatc c
//
