LOCUS       Exported                7871 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pSHTB10, complete sequence.
ACCESSION   KY793908
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7871)
  AUTHORS   Wang Y, Su H, Li G, Yang M, Yi J.
  TITLE     Direct Submission
  JOURNAL   Submitted (21-MAR-2017) School of Environmental and Biological 
            Engineering, Nanjing University of Science and Technology, 
            XiaoLinwei 200#, Nan Jing, JiangSu 210094, China
REFERENCE   2  (bases 1 to 7871)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7871
                     /organism="Cloning vector pSHTB10"
                     /mol_type="other DNA"
                     /db_xref="taxon:2093925"
     misc_feature    1..504
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     rep_origin      1..492
                     /note="CEN6/ARSH4; derived from Saccharomyces cerevisiae"
     promoter        774..989
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             990..1793
                     /codon_start=1
                     /gene="Ura3"
                     /product="Ura3"
                     /label=Ura3
                     /note="selectable marker in Saccharomyces cerevisiae"
                     /protein_id="AVI16869.1"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     gene            990..1793
                     /gene="Ura3"
                     /label=Ura3
     rep_origin      complement(1924..2379)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     2524..2540
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2550..2568
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2577..2835
                     /regulatory_class="terminator"
     terminator      2587..2834
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     CDS             complement(3376..4089)
                     /codon_start=1
                     /gene="I-SceI"
                     /product="intron-encoded endonuclease"
                     /label=I-SceI
                     /protein_id="AVI16870.1"
                     /translation="MHMKNIKKNQVMNLGPNSKLLKEYKSQLIELNIEQFEAGIGLILG
                     DAYIRSRDEGKTYCMQFEWKNKAYMDHVCLLYDQWVLSPPHKKERVNHLGNLVITWGAQ
                     TFKHQAFNKLANLFIVNNKKTIPNNLVENYLTPMSLAYWFMDDGGKWDYNKNSTNKSIV
                     LNTQSFTFEEVEYLVKGLRNKFQLNCYVKINKNKPIIYIDSMSYLIFYNLIKPYLIPQM
                     MYKLPNTISSETFLK"
     gene            complement(3376..4089)
                     /gene="I-SceI"
                     /label=I-SceI
     protein_bind    4090..4113
                     /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     complement(4147..4163)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      complement(4158..4625)
                     /regulatory_class="promoter"
                     /note="Gal1-V10 promoter"
     promoter        complement(4178..4619)
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    4502..4619
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     rep_origin      4650..5175
                     /label=oriV
                     /note="oriV"
     rep_origin      4651..5181
                     /label=oriV
                     /note="incP origin of replication"
     regulatory      5403..5521
                     /regulatory_class="promoter"
                     /note="neo promoter"
     CDS             5573..6721
                     /codon_start=1
                     /gene="trfA"
                     /product="replication protein"
                     /label=trfA
                     /protein_id="AVI16871.1"
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFTWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     gene            5573..6721
                     /gene="trfA"
                     /label=trfA
     regulatory      6772..7377
                     /regulatory_class="terminator"
     regulatory      7381..7434
                     /regulatory_class="promoter"
                     /note="hbs promoter"
     CDS             7435..7809
                     /codon_start=1
                     /gene="BleR"
                     /product="bleomycin resistance protein"
                     /label=BleR
                     /protein_id="AVI16872.1"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     gene            7435..7809
                     /gene="BleR"
                     /label=BleR