LOCUS       Exported                9742 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  YAC construction vector pRML1, complete sequence.
ACCESSION   EF036510
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9742)
  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 9742)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9742
                     /organism="YAC construction vector pRML1"
                     /mol_type="other DNA"
                     /db_xref="taxon:410732"
     regulatory      complement(50..69)
                     /regulatory_class="promoter"
                     /note="T3 promoter; primer"
     promoter        complement(52..70)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    100..956
                     /label=centromere region from yeast chromosome 4
                     /note="centromere region from yeast chromosome 4"
     misc_feature    650..761
                     /label=centromere from yeast chromosome 4
                     /note="centromere from yeast chromosome 4"
     misc_feature    664..956
                     /label=CEN4
                     /note="S. cerevisiae CEN4 centromere"
     regulatory      complement(961..1416)
                     /regulatory_class="promoter"
                     /note="Gal1 promoter oriented to influence centromere 
                     activity"
     promoter        complement(973..1414)
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    1297..1414
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     misc_feature    1544..3607
                     /note="ARS/Trp1 region from chromosome 4; contains full 
                     promoter for Trp1 gene unlike pYAC4 sequence."
     misc_feature    1548..2386
                     /label=ARS
                     /note="ARS"
     rep_origin      1548..2385
                     /label=ARS1
                     /note="S. cerevisiae autonomously replicating sequence 
                     ARS1/ARS416"
     CDS             complement(2224..2898)
                     /codon_start=1
                     /product="Trp1"
                     /label=Trp1
                     /note="N-5 phosphoribosyl anthranilate isomerase"
                     /protein_id="ABK41442.1"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     promoter        complement(2899..3180)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     promoter        3922..3984
                     /label=Mini-TK promoter
                     /note="minimal herpes simplex virus (HSV) thymidine kinase 
                     promoter"
     misc_feature    4744..7035
                     /label=thymidine kinase region from Herpesvirus
                     /note="thymidine kinase region from Herpesvirus"
     5UTR           4747..4786
                     /label=HSV TK 5-UTR
                     /note="5 untranslated region from the herpes simplex virus
                     thymidine kinase gene"
     CDS             4804..5934
                     /codon_start=1
                     /product="thymidine kinase"
                     /label=thymidine kinase
                     /protein_id="ABK41443.1"
                     /translation="MASYPCHQHASAFDQAARSRGHNNRRTALRPRRQQKATEVRLEQK
                     MPTLLRVYIDGPHGMGKTTTTQLLVALGSRDDIVYVPEPMTYWRVLGASETIANIYTTQ
                     HRLDQGEISAGDAAVVMTSAQITMGMPYAVTDAVLAPHIGGEAGSSHAPPPALTLIFDR
                     HPIAALLCYPAARYLMGSMTPQAVLAFVALIPPTLPGTNIVLGALPEDRHIDRLAKRQR
                     PGERLDLAMLAAIRRVYGLLANTVRYLQGGGSWREDWGQLSGAAVPPQGAEPQSNAGPR
                     PHIGDTLFTLFRAPELLAPNGDLYNVFAWALDVLAKRLRPMHVFILDYDQSPAGCRDAL
                     LQLTSGMVQTHVTTPGSIPTICDLARTFAREMGEAN"
     polyA_signal    5976..6024
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     misc_feature    7038..8972
                     /label=pBluescript II SK+ region
                     /note="pBluescript II SK+ region"
     promoter        7061..7165
                     /gene="bla"
                     /label=AmpR promoter
     CDS             7166..8026
                     /codon_start=1
                     /product="bla"
                     /label=bla
                     /note="beta-lactamase"
                     /protein_id="ABK41444.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      8197..8785
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    9097..9723
                     /note="telomere region from pYAC4 of GenBank Accession 
                     Number U01086"