LOCUS       40924_29566        3626 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pLZ44, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3626)
  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 3626)
  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 3626)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 3626)
  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..3626
                     /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)."
     CDS             467..1180
                     /codon_start=1
                     /label=yeGFP
                     /note="yeast-enhanced green fluorescent protein"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFAYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      1267..1513
                     /label=lambda tL3 terminator
                     /note="transcription terminator tL3 from phage lambda"
     protein_bind    1527..1844
                     /label=phage Phi-80 attP
                     /note="attachment site of phage phi-80"
     rep_origin      complement(1955..2343)
                     /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(2494..3150)
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPKFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     promoter        complement(3151..3253)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     misc_feature    3450..3626
                     /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 atacatatga gtaaaggaga
      481 agaacttttc actggagttg tcccaattct tgttgaatta gatggtgatg ttaatgggca
      541 caaattttct gtcagtggag agggtgaagg tgatgcaaca tacggaaaac ttacccttaa
      601 atttatttgc actactggaa aactacctgt tccatggcca acacttgtca ctactttcgc
      661 gtatggtctt caatgctttg cgagataccc agatcatatg aaacagcatg actttttcaa
      721 gagtgccatg cccgaaggtt atgtacagga aagaactata tttttcaaag atgacgggaa
      781 ctacaagaca cgtgctgaag tcaagtttga aggtgatacc cttgttaata gaatcgagtt
      841 aaaaggtatt gattttaaag aagatggaaa cattcttgga cacaaattgg aatacaacta
      901 taactcacac aatgtataca tcatggcaga caaacaaaag aatggaatca aagttaactt
      961 caaaattaga cacaacattg aagatggaag cgttcaacta gcagaccatt atcaacaaaa
     1021 tactccaatt ggcgatggcc ctgtcctttt accagacaac cattacctgt ccacacaatc
     1081 tgccctttcg aaagatccca acgaaaagag agaccacatg gtccttcttg agtttgtaac
     1141 agctgctggg attacacatg gcatggatga actatacaaa taacgaagag ctgcaggtcg
     1201 acggatcccc ggaattaatt ctcatgtttg acagcttatc actgatcagt gaattaatgg
     1261 cgatgacgca tcctcacgat aatatccggg taggcgcaat cactttcgtc tctactccgt
     1321 tacaaagcga ggctgggtat ttcccggcct ttctgttatc cgaaatccac tgaaagcaca
     1381 gcggctggct gaggagataa ataataaacg aggggctgta tgcacaaagc atcttctgtt
     1441 gagttaagaa cgagtatcga gatggcacat agccttgctc aaattggaat caggtttgtg
     1501 ccaataccag tagaaacaga cgaagaagct agctgataac gggaaagaaa cagagaaggg
     1561 cacaaatatt gtgtacttta atgtgccctt taatttattg attggtggtt gaattgtccg
     1621 taactttttg atttaagtgc aaatttctaa taaattagaa cactttctta aattgtcatt
     1681 tggcatatta cgaacaattc cgcgtaaaaa cgttctgtta cgctaaaccc ttatccagca
     1741 ggctttcaag gatgtaaacc ataacactct gcgaactagt gttacattgc gtgtagcttt
     1801 gagtgggcaa ctttgtgtac acttttgtgt acccaaaaac aaaaatgtgt acccattcaa
     1861 tgatcaccga cacaaagctc aggaaggcgc tcggcaagaa aagagatgat atcgagatta
     1921 tttctgattc gcacgggccc atggctaatt cccatgtcag ccgttaagtg ttcctgtgtc
     1981 actgaaaatt gctttgagag gctctaaggg cttctcagtg cgttacatcc ctggcttgtt
     2041 gtccacaacc gttaaacctt aaaagcttta aaagccttat atattctttt ttttcttata
     2101 aaacttaaaa ccttagaggc tatttaagtt gctgatttat attaatttta ttgttcaaac
     2161 atgagagctt agtacgtgaa acatgagagc ttagtacgtt agccatgaga gcttagtacg
     2221 ttagccatga gggtttagtt cgttaaacat gagagcttag tacgttaaac atgagagctt
     2281 agtacgtgaa acatgagagc ttagtacgta ctatcaacag gttgaactgc tgatcttcag
     2341 atcctctacg ccggacgcat cgtggccgga tcttgcggcc gcaaaaatta aaaatgaagt
     2401 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     2461 agtgaggcac caataactgc cttaaaaaaa ttacgccccg ccctgccact catcgcagta
     2521 ctgttgtaat tcattaagca ttctgccgac atggaagcca tcacagacgg catgatgaac
     2581 ctgaatcgcc agcggcatca gcaccttgtc gccttgcgta taatatttgc ccatggtgaa
     2641 aacgggggcg aagaagttgt ccatattggc cacgtttaaa tcaaaactgg tgaaactcac
     2701 ccagggattg gctgagacga aaaacatatt ctcaataaac cctttaggga aataggccag
     2761 gttttcaccg taacacgcca catcttgcga atatatgtgt agaaactgcc ggaaatcgtc
     2821 gtggtattca ctccagagcg atgaaaacgt ttcagtttgc tcatggaaaa cggtgtaaca
     2881 agggtgaaca ctatcccata tcaccagctc accgtctttc attgccatac ggaatttcgg
     2941 atgagcattc atcaggcggg caagaatgtg aataaaggcc ggataaaact tgtgcttatt
     3001 tttctttacg gtctttaaaa aggccgtaat atccagctga acggtctggt tataggtaca
     3061 ttgagcaact gactgaaatg cctcaaaatg ttctttacga tgccattggg atatatcaac
     3121 ggtggtatat ccagtgattt ttttctccat tttagcttcc ttagctcctg aaaatctcga
     3181 taactcaaaa aatacgcccg gtagtgatct tatttcatta tggtgaaagt tggaacctct
     3241 tacgtgccga tcaacgtctc attttcgcca aaagttggcc cagggcttcc cggtatcaac
     3301 agggacacca ggatttattt attctgcgaa gtgatcttcc gtcacaggta tttattcggc
     3361 gcaaagtgcg tcgggtgatg ctgccaactt actgatttag tgtatgatgg tgtttttgag
     3421 gtgctccagt ggcttctgtt tctatcagca tcgatgatta tagcgatttt ttcggtatat
     3481 ccatcctttt tcgcacgata tacaggattt tgccaaaggg ttcgtgtaga ctttccttgg
     3541 tgtatccaac ggcgtcagcc gggcaggata ggtgaagtag gcccacccgc gagcgggtgt
     3601 tccttcttca ctgtccctta ttcgca
//
