LOCUS       Exported                5593 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pMAK705, complete sequence.
ACCESSION   AF519766
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5593)
  AUTHORS   Hamilton CM, Aldea M, Washburn BK, Babitzke P, Kushner SR.
  TITLE     New method for generating deletions and gene replacements in 
            Escherichia coli
  JOURNAL   J. Bacteriol. 171 (9), 4617-4622 (1989)
  PUBMED    2548993
REFERENCE   2  (bases 1 to 5593)
  AUTHORS   Maples VF, Kushner SR.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-JUN-2002) Genetics, University of Georgia, Life 
            Sciences Bldg., Athens, GA 30602, USA
REFERENCE   3  (bases 1 to 5593)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5593
                     /organism="Cloning vector pMAK705"
                     /plasmid="pMAK705"
                     /mol_type="genomic DNA"
                     /note="designed for chromosomal deletion and gene 
                     replacement"
                     /db_xref="taxon:198575"
     misc_feature    8..2937
                     /note="derived from vector pSC101 deposited in EMBL 
                     Accession Number X01654"
     rep_origin      487..709
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             757..1707
                     /codon_start=1
                     /gene="rep"
                     /product="rep101"
                     /label=rep
                     /note="involved in replication of plasmids pHS1 and pSC101"
                     /protein_id="AAM66744.1"
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYVQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     gene            757..1707
                     /gene="rep"
                     /label=rep
     misc_feature    2943..5065
                     /note="derived from vector pBR325 depsited in GenBank 
                     Accession Number L08855"
     promoter        complement(3191..3295)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3498..3600
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             3601..4260
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=cat
                     /note="confers chloramphenicol resistance"
                     /protein_id="AAM66745.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     gene            3601..4260
                     /gene="cat"
                     /label=cat
     CDS             complement(5065..5295)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGE"
     misc_feature    5066..5505
                     /note="derived from vector pUC19 deposited in GenBank 
                     Accession Number L09137"
     primer_bind     5205..5221
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    5222..5278
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(5291..5307)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5315..5331
                     /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)."
     promoter        complement(5339..5369)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5384..5405
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_feature    5506..5593
                     /note="derived from vector pBR325 deposited in GenBank 
                     Accession Number L08855"