LOCUS       Exported                3626 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pLZ44, complete sequence.
ACCESSION   AY236526
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)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..3626
                     /organism="Cloning vector pLZ44"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:234844"
     misc_feature    1..260
                     /label=rgnB
                     /note="rgnB"
     terminator      50..136
                     /gene="Escherichia coli rrnB"
                     /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"
     regulatory      283..464
                     /regulatory_class="promoter"
                     /note="lac UV5"
     protein_bind    357..376
                     /label=lac operator (symmetric)
                     /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). 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
                     /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)."
     CDS             467..1183
                     /codon_start=1
                     /product="Gfp"
                     /label=Gfp
                     /note="mut2"
                     /protein_id="AAP74499.1"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFAYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     misc_feature    1256..1525
                     /label=tL3
                     /note="tL3"
     terminator      1267..1513
                     /label=-lambda tL3 terminator
                     /note="transcription terminator tL3 from phage lambda"
     protein_bind    1527..1844
                     /label=phage Phi-80 attP
                     /bound_moiety="phage phi-80 integrase"
                     /note="attachment site of phage phi-80"
     misc_feature    1529..1935
                     /label=att80
                     /note="att80"
     misc_feature    1943..2375
                     /label=oriRg
                     /note="oriRg"
     rep_origin      1955..2343
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     CDS             complement(2491..3150)
                     /codon_start=1
                     /product="Cat"
                     /label=Cat
                     /protein_id="AAP74500.1"
                     /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"