LOCUS       40924_29561        3729 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pLZ43, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3729)
  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 3729)
  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 3729)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 3729)
  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..3729
                     /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"
     promoter        425..543
                     /label=rhaB promoter
                     /note="promoter of the E. coli rhaBAD operon, conferring
                     tight induction with L-rhamnose and repression with 
                     D-glucose in the presence of RhaR and RhaS (Giacalone et 
                     al., 2006)"
     CDS             573..1286
                     /codon_start=1
                     /label=yeGFP
                     /note="yeast-enhanced green fluorescent protein"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFAYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      1370..1616
                     /label=lambda tL3 terminator
                     /note="transcription terminator tL3 from phage lambda"
     protein_bind    1630..1947
                     /label=phage Phi-80 attP
                     /note="attachment site of phage phi-80"
     rep_origin      complement(2058..2446)
                     /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(2597..3253)
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPKFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     promoter        complement(3254..3356)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     misc_feature    3553..3729
                     /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 gcggcctcct gatgtcgtca
      301 acacggcgaa atagtaatca cgaggtcagg ttcttacctt aaattttcga cggaaaacca
      361 cgtaaaaaac gtcgattttt caagatacag cgtgaatttt caggaaatgc ggtgagcatc
      421 acatcaccac aattcagcaa attgtgaaca tcatcacgtt catctttccc tggttgccaa
      481 tggcccattt tcctgtcagt aacgagaagg tcgcgaattc aggcgctttt tagactggtc
      541 gtaatgaaat tcagcaggat cacactagtc atatgagtaa aggagaagaa cttttcactg
      601 gagttgtccc aattcttgtt gaattagatg gtgatgttaa tgggcacaaa ttttctgtca
      661 gtggagaggg tgaaggtgat gcaacatacg gaaaacttac ccttaaattt atttgcacta
      721 ctggaaaact acctgttcca tggccaacac ttgtcactac tttcgcgtat ggtcttcaat
      781 gctttgcgag atacccagat catatgaaac agcatgactt tttcaagagt gccatgcccg
      841 aaggttatgt acaggaaaga actatatttt tcaaagatga cgggaactac aagacacgtg
      901 ctgaagtcaa gtttgaaggt gatacccttg ttaatagaat cgagttaaaa ggtattgatt
      961 ttaaagaaga tggaaacatt cttggacaca aattggaata caactataac tcacacaatg
     1021 tatacatcat ggcagacaaa caaaagaatg gaatcaaagt taacttcaaa attagacaca
     1081 acattgaaga tggaagcgtt caactagcag accattatca acaaaatact ccaattggcg
     1141 atggccctgt ccttttacca gacaaccatt acctgtccac acaatctgcc ctttcgaaag
     1201 atcccaacga aaagagagac cacatggtcc ttcttgagtt tgtaacagct gctgggatta
     1261 cacatggcat ggatgaacta tacaaataac gaagagcgga tccccgggta ccgagctcga
     1321 attctcatgt ttgacagctt atcactgatc agtgaattaa tggcgatgac gcatcctcac
     1381 gataatatcc gggtaggcgc aatcactttc gtctctactc cgttacaaag cgaggctggg
     1441 tatttcccgg cctttctgtt atccgaaatc cactgaaagc acagcggctg gctgaggaga
     1501 taaataataa acgaggggct gtatgcacaa agcatcttct gttgagttaa gaacgagtat
     1561 cgagatggca catagccttg ctcaaattgg aatcaggttt gtgccaatac cagtagaaac
     1621 agacgaagaa gctagctgat aacgggaaag aaacagagaa gggcacaaat attgtgtact
     1681 ttaatgtgcc ctttaattta ttgattggtg gttgaattgt ccgtaacttt ttgatttaag
     1741 tgcaaatttc taataaatta gaacactttc ttaaattgtc atttggcata ttacgaacaa
     1801 ttccgcgtaa aaacgttctg ttacgctaaa cccttatcca gcaggctttc aaggatgtaa
     1861 accataacac tctgcgaact agtgttacat tgcgtgtagc tttgagtggg caactttgtg
     1921 tacacttttg tgtacccaaa aacaaaaatg tgtacccatt caatgatcac cgacacaaag
     1981 ctcaggaagg cgctcggcaa gaaaagagat gatatcgaga ttatttctga ttcgcacggg
     2041 cccatggcta attcccatgt cagccgttaa gtgttcctgt gtcactgaaa attgctttga
     2101 gaggctctaa gggcttctca gtgcgttaca tccctggctt gttgtccaca accgttaaac
     2161 cttaaaagct ttaaaagcct tatatattct tttttttctt ataaaactta aaaccttaga
     2221 ggctatttaa gttgctgatt tatattaatt ttattgttca aacatgagag cttagtacgt
     2281 gaaacatgag agcttagtac gttagccatg agagcttagt acgttagcca tgagggttta
     2341 gttcgttaaa catgagagct tagtacgtta aacatgagag cttagtacgt gaaacatgag
     2401 agcttagtac gtactatcaa caggttgaac tgctgatctt cagatcctct acgccggacg
     2461 catcgtggcc ggatcttgcg gccgcaaaaa ttaaaaatga agttttaaat caatctaaag
     2521 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg caccaataac
     2581 tgccttaaaa aaattacgcc ccgccctgcc actcatcgca gtactgttgt aattcattaa
     2641 gcattctgcc gacatggaag ccatcacaga cggcatgatg aacctgaatc gccagcggca
     2701 tcagcacctt gtcgccttgc gtataatatt tgcccatggt gaaaacgggg gcgaagaagt
     2761 tgtccatatt ggccacgttt aaatcaaaac tggtgaaact cacccaggga ttggctgaga
     2821 cgaaaaacat attctcaata aaccctttag ggaaataggc caggttttca ccgtaacacg
     2881 ccacatcttg cgaatatatg tgtagaaact gccggaaatc gtcgtggtat tcactccaga
     2941 gcgatgaaaa cgtttcagtt tgctcatgga aaacggtgta acaagggtga acactatccc
     3001 atatcaccag ctcaccgtct ttcattgcca tacggaattt cggatgagca ttcatcaggc
     3061 gggcaagaat gtgaataaag gccggataaa acttgtgctt atttttcttt acggtcttta
     3121 aaaaggccgt aatatccagc tgaacggtct ggttataggt acattgagca actgactgaa
     3181 atgcctcaaa atgttcttta cgatgccatt gggatatatc aacggtggta tatccagtga
     3241 tttttttctc cattttagct tccttagctc ctgaaaatct cgataactca aaaaatacgc
     3301 ccggtagtga tcttatttca ttatggtgaa agttggaacc tcttacgtgc cgatcaacgt
     3361 ctcattttcg ccaaaagttg gcccagggct tcccggtatc aacagggaca ccaggattta
     3421 tttattctgc gaagtgatct tccgtcacag gtatttattc ggcgcaaagt gcgtcgggtg
     3481 atgctgccaa cttactgatt tagtgtatga tggtgttttt gaggtgctcc agtggcttct
     3541 gtttctatca gcatcgatga ttatagcgat tttttcggta tatccatcct ttttcgcacg
     3601 atatacagga ttttgccaaa gggttcgtgt agactttcct tggtgtatcc aacggcgtca
     3661 gccgggcagg ataggtgaag taggcccacc cgcgagcggg tgttccttct tcactgtccc
     3721 ttattcgca
//
