LOCUS       Exported                7144 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pLM10, complete sequence.
ACCESSION   AF108217
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7144)
  AUTHORS   Marino-Ramirez L, Blum BJ, Hu JC.
  TITLE     Green fluorescent protein (GFP) as a reporter to map protein-protein
            interactions in Escherichia coli
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7144)
  AUTHORS   Marino-Ramirez L, Blum BJ, Hu JC.
  TITLE     Direct Submission
  JOURNAL   Submitted (19-NOV-1998) Biochemistry & Biophysics, Texas A&M 
            University, College Station, TX 77843-2128, USA
REFERENCE   3  (bases 1 to 7144)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7144
                     /organism="Cloning vector pLM10"
                     /host="Escherichia coli AG1688"
                     /mol_type="other DNA"
                     /db_xref="taxon:89245"
     CDS             2650..2841
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     misc_feature    2943..3083
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(3269..3857)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4028..4888)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /protein_id="AAD25334.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(4028..4888)
                     /gene="bla"
                     /label=bla
     promoter        complement(4889..4980)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      4999..5026
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      5118..5204
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(5500..6159)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=cat
                     /protein_id="AAD25335.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     gene            complement(5500..6159)
                     /gene="cat"
                     /label=cat
     regulatory      complement(6212..6269)
                     /regulatory_class="promoter"
                     /note="lacUV5 promoter without operator"
                     /experiment="experimental evidence, no additional details 
                     recorded"
     promoter        complement(6217..6247)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     gene            6276..7132
                     /gene="gfp"
                     /allele="mut2"
                     /label=gfp
                     /experiment="experimental evidence, no additional details 
                     recorded"
     regulatory      6276..6330
                     /regulatory_class="promoter"
                     /gene="gfp"
                     /allele="mut2"
                     /note="pL promoter"
                     /experiment="experimental evidence, no additional details 
                     recorded"
     misc_binding    6281..6295
                     /gene="gfp"
                     /allele="mut2"
                     /label=O-L2 repressor binding site
                     /bound_moiety="O-L2 repressor"
     misc_binding    6305..6321
                     /gene="gfp"
                     /allele="mut2"
                     /label=O-L1 repressor binding site
                     /bound_moiety="O-L1 repressor"
     CDS             6376..7092
                     /codon_start=1
                     /gene="gfp"
                     /allele="mut2"
                     /product="green fluorescent protein"
                     /label=gfp
                     /note="optimized for FACS"
                     /protein_id="AAD25336.1"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFAYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     regulatory      7109..7132
                     /regulatory_class="attenuator"
                     /gene="gfp"
                     /allele="mut2"