LOCUS       Exported                5180 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  YAC construction vector pRML2, complete sequence.
ACCESSION   EF036511
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5180)
  AUTHORS   Knight TF Jr.
  TITLE     Direct Submission
  JOURNAL   Submitted (03-OCT-2006) CSAIL, MIT, 32 Vassar St, Cambridge, MA 
            02139, USA
REFERENCE   2  (bases 1 to 5180)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5180
                     /organism="YAC construction vector pRML2"
                     /mol_type="other DNA"
                     /db_xref="taxon:410733"
     regulatory      67..86
                     /regulatory_class="promoter"
                     /note="T7 promoter; primer"
     promoter        complement(68..86)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    104..485
                     /label=ARSH4 region
                     /note="ARSH4 region"
     misc_feature    complement(489..1649)
                     /label=Ura3 cassette
                     /note="Ura3 cassette"
     CDS             complement(630..1433)
                     /codon_start=1
                     /product="Ura3"
                     /label=Ura3
                     /protein_id="ABK41445.1"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(1434..1649)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     misc_feature    1671..4462
                     /label=pBluescript II SK+ region
                     /note="pBluescript II SK+ region"
     primer_bind     complement(1678..1694)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      1836..2291
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        2317..2421
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2422..3282
                     /codon_start=1
                     /product="bla"
                     /label=bla
                     /note="beta-lactamase"
                     /protein_id="ABK41446.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3453..4041
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    4329..4350
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4365..4395
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4403..4419
                     /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)."
     primer_bind     4427..4443
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4469..4485
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    4529..5161
                     /note="telomere region from pYAC4 of GenBank Accession 
                     Number U01086"