LOCUS       Exported                5634 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pRS413-SUP61S, complete sequence.
ACCESSION   KF214847
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5634)
  AUTHORS   Muller HM, Annaluru N, Boeke JD, Chandrasegaran S.
  TITLE     Total Synthesis of a Functional Designer Eukaryotic Chromosome
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5634)
  AUTHORS   Muller HM, Annaluru N, Boeke JD, Chandrasegaran S.
  TITLE     Direct Submission
  JOURNAL   Submitted (10-JUN-2013) Genomes et Genetique, Institut Pasteur, 25 
            rue du Docteur Roux, Paris 75015, France
REFERENCE   3  (bases 1 to 5634)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..5634
                     /organism="Cloning vector pRS413-SUP61S"
                     /mol_type="other DNA"
                     /label=derived from Saccharomyces cerevisiae
                     /PCR_primers="fwd_name: m13-for, fwd_seq:
                     tgtaaaacgacggccagt, fwd_name: t7, fwd_seq: 
                     gtaatacgactcactataggg, rev_name: m13-rev, rev_seq: 
                     caggaaacagctatgaccatg, rev_name: t3, rev_seq: 
                     attaaccctcactaaaggga"
                     /note="derived from Saccharomyces cerevisiae"
                     /db_xref="taxon:1401519"
     misc_feature    193..1364
                     /label=HIS3 cassette
                     /note="HIS3 cassette"
     promoter        317..503
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             504..1163
                     /codon_start=1
                     /product="HIS3"
                     /label=HIS3
                     /protein_id="AGW18140.1"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALLARGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHFL
                     ESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     rep_origin      1423..1878
                     /label=M13 origin
                     /note="M13 origin"
     misc_feature    1883..1951
                     /label=LacZ alpha
                     /note="LacZ alpha"
     primer_bind     2023..2039
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2049..2067
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     2093..2109
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    2121..2784
                     /label=synSUP61 cassette
                     /note="synSUP61 cassette"
     primer_bind     complement(2807..2823)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(2860..2878)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2899..2915)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    2921..2943
                     /label=LacO
                     /note="LacO"
     protein_bind    2923..2939
                     /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(2947..2977)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     rep_origin      3283..3965
                     /label=colE1 origin
                     /note="colE1 origin"
     rep_origin      complement(3301..3889)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4060..4920)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     CDS             complement(4060..4722)
                     /codon_start=1
                     /product="AmpR"
                     /label=AmpR
                     /protein_id="AGW18141.1"
                     /translation="MSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHL
                     TDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEA
                     IPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFI
                     ADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW"
     promoter        complement(4921..5025)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    5052..5562
                     /label=ARS-CEN pRS
                     /note="ARS-CEN pRS"
     misc_feature    5062..5565
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"