LOCUS       40924_29556        3890 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pLZ42, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3890)
  AUTHORS   Zhou L, Zhang K, Wanner BL.
  TITLE     Chromosomal expression of foreign and native genes from regulatable 
            promoters in Escherichia coli
  JOURNAL   Methods Mol. Biol. 267, 123-134 (2004)
  PUBMED    15269420
REFERENCE   2  (bases 1 to 3890)
  AUTHORS   Zhou L, Zhang K, Wanner BL.
  TITLE     Direct Submission
  JOURNAL   Submitted (13-FEB-2003) Biological Sciences, Purdue University, 
            Lilly Hall, West Lafayette, IN 47907, USA
REFERENCE   3  (bases 1 to 3890)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 3890)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Methods 
            Mol. Biol. 267, 123-134 (2004)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (13-FEB-2003) Biological Sciences, Purdue University, Lilly Hall, 
            West Lafayette, IN 47907, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..3890
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      50..136
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      228..255
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     protein_bind    complement(357..376)
                     /label=lac operator (symmetric)
                     /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). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     promoter        391..421
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     protein_bind    429..445
                     /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)."
     promoter        475..579
                     /label=AmpR promoter
     CDS             580..1437
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     terminator      1531..1777
                     /label=lambda tL3 terminator
                     /note="transcription terminator tL3 from phage lambda"
     protein_bind    1791..2108
                     /label=phage Phi-80 attP
                     /note="attachment site of phage phi-80"
     rep_origin      complement(2219..2607)
                     /direction=LEFT
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     CDS             complement(2758..3414)
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPKFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     promoter        complement(3415..3517)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     misc_feature    3714..3890
                     /label=oriT2
                     /note="oriT2"
ORIGIN
        1 gatggtagtg tggggtctcc ccatgcgaga gtagggaact gccaggcatc aaataaaacg
       61 aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct
      121 cctgagtagg acaaatccgc cgggagcgga tttgaacgtt gcgaagcaac ggcccggagg
      181 gtggcgggca ggacgcccgc cataaactgc caggcatcaa attaagcaga aggccatcct
      241 gacggatggc ctttttgcgt ggccagtgcc aagcttgcat gcagatctag cgctctagag
      301 gatctgcggc cgcatttttg cggccttttt ttttggatct actagtgcta gatcgaaatt
      361 gtgagcgctc acaattggct agctcgaggc tttacacttt atgcttccgg ctcgtataat
      421 gtgtggaatt gtgagcggat aacaatttca cacaggagat atacatatgt gaagagcgga
      481 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
      541 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
      601 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
      661 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
      721 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
      781 agcactttta aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag
      841 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
      901 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
      961 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     1021 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     1081 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     1141 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     1201 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     1261 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     1321 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     1381 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     1441 ctgttaacga agagctgcag gtcgacggat ccccggaatt aattctcatg tttgacagct
     1501 tatcactgat cagtgaatta atggcgatga cgcatcctca cgataatatc cgggtaggcg
     1561 caatcacttt cgtctctact ccgttacaaa gcgaggctgg gtatttcccg gcctttctgt
     1621 tatccgaaat ccactgaaag cacagcggct ggctgaggag ataaataata aacgaggggc
     1681 tgtatgcaca aagcatcttc tgttgagtta agaacgagta tcgagatggc acatagcctt
     1741 gctcaaattg gaatcaggtt tgtgccaata ccagtagaaa cagacgaaga agctagctga
     1801 taacgggaaa gaaacagaga agggcacaaa tattgtgtac tttaatgtgc cctttaattt
     1861 attgattggt ggttgaattg tccgtaactt tttgatttaa gtgcaaattt ctaataaatt
     1921 agaacacttt cttaaattgt catttggcat attacgaaca attccgcgta aaaacgttct
     1981 gttacgctaa acccttatcc agcaggcttt caaggatgta aaccataaca ctctgcgaac
     2041 tagtgttaca ttgcgtgtag ctttgagtgg gcaactttgt gtacactttt gtgtacccaa
     2101 aaacaaaaat gtgtacccat tcaatgatca ccgacacaaa gctcaggaag gcgctcggca
     2161 agaaaagaga tgatatcgag attatttctg attcgcacgg gcccatggct aattcccatg
     2221 tcagccgtta agtgttcctg tgtcactgaa aattgctttg agaggctcta agggcttctc
     2281 agtgcgttac atccctggct tgttgtccac aaccgttaaa ccttaaaagc tttaaaagcc
     2341 ttatatattc ttttttttct tataaaactt aaaaccttag aggctattta agttgctgat
     2401 ttatattaat tttattgttc aaacatgaga gcttagtacg tgaaacatga gagcttagta
     2461 cgttagccat gagagcttag tacgttagcc atgagggttt agttcgttaa acatgagagc
     2521 ttagtacgtt aaacatgaga gcttagtacg tgaaacatga gagcttagta cgtactatca
     2581 acaggttgaa ctgctgatct tcagatcctc tacgccggac gcatcgtggc cggatcttgc
     2641 ggccgcaaaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     2701 tctgacagtt accaatgctt aatcagtgag gcaccaataa ctgccttaaa aaaattacgc
     2761 cccgccctgc cactcatcgc agtactgttg taattcatta agcattctgc cgacatggaa
     2821 gccatcacag acggcatgat gaacctgaat cgccagcggc atcagcacct tgtcgccttg
     2881 cgtataatat ttgcccatgg tgaaaacggg ggcgaagaag ttgtccatat tggccacgtt
     2941 taaatcaaaa ctggtgaaac tcacccaggg attggctgag acgaaaaaca tattctcaat
     3001 aaacccttta gggaaatagg ccaggttttc accgtaacac gccacatctt gcgaatatat
     3061 gtgtagaaac tgccggaaat cgtcgtggta ttcactccag agcgatgaaa acgtttcagt
     3121 ttgctcatgg aaaacggtgt aacaagggtg aacactatcc catatcacca gctcaccgtc
     3181 tttcattgcc atacggaatt tcggatgagc attcatcagg cgggcaagaa tgtgaataaa
     3241 ggccggataa aacttgtgct tatttttctt tacggtcttt aaaaaggccg taatatccag
     3301 ctgaacggtc tggttatagg tacattgagc aactgactga aatgcctcaa aatgttcttt
     3361 acgatgccat tgggatatat caacggtggt atatccagtg atttttttct ccattttagc
     3421 ttccttagct cctgaaaatc tcgataactc aaaaaatacg cccggtagtg atcttatttc
     3481 attatggtga aagttggaac ctcttacgtg ccgatcaacg tctcattttc gccaaaagtt
     3541 ggcccagggc ttcccggtat caacagggac accaggattt atttattctg cgaagtgatc
     3601 ttccgtcaca ggtatttatt cggcgcaaag tgcgtcgggt gatgctgcca acttactgat
     3661 ttagtgtatg atggtgtttt tgaggtgctc cagtggcttc tgtttctatc agcatcgatg
     3721 attatagcga ttttttcggt atatccatcc tttttcgcac gatatacagg attttgccaa
     3781 agggttcgtg tagactttcc ttggtgtatc caacggcgtc agccgggcag gataggtgaa
     3841 gtaggcccac ccgcgagcgg gtgttccttc ttcactgtcc cttattcgca
//
