LOCUS       Exported               12084 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pDEST374-U1Ux, complete sequence.
ACCESSION   KX344515
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12084)
  AUTHORS   MacPherson MI, Saka Y.
  TITLE     Short Synthetic Terminators for Assembly of Transcription Units in 
            vitro and Stable Chromosomal Integration in Yeast S. cerevisiae
  JOURNAL   ACS Synth Biol (2016) In press
  PUBMED    27529501
REFERENCE   2  (bases 1 to 12084)
  AUTHORS   Saka Y.
  TITLE     Direct Submission
  JOURNAL   Submitted (27-MAY-2016) School of Medicine, Medical Sciences & 
            Nutrition, Institute of Medical Sciences, University of Aberdeen, 
            Foresterhill, Aberdeen AB25 2ZD, UK
REFERENCE   3  (bases 1 to 12084)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..12084
                     /organism="Cloning vector pDEST374-U1Ux"
                     /mol_type="other DNA"
                     /db_xref="taxon:1892301"
     misc_feature    complement(71..466)
                     /label=HIS3 YIp-In
                     /note="HIS3 YIp-In"
     misc_feature    474..513
                     /label=U1 sequence
                     /note="U1 sequence"
     misc_feature    530..569
                     /label=Ux sequence
                     /note="Ux sequence"
     misc_feature    complement(587..1130)
                     /label=HIS3 YIp-Out
                     /note="HIS3 YIp-Out"
     promoter        complement(894..1079)
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             1138..1155
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     regulatory      1222..1269
                     /regulatory_class="terminator"
                     /note="T7 terminator"
     terminator      1222..1269
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1643..2521)
                     /codon_start=1
                     /gene="araC"
                     /product="AraC protein"
                     /label=araC
                     /protein_id="AOF43193.1"
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     gene            complement(1643..2521)
                     /gene="araC"
                     /label=araC
     promoter        2548..2832
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     regulatory      2672..2824
                     /regulatory_class="promoter"
                     /note="Arabinose promoter"
     CDS             2892..4040
                     /codon_start=1
                     /gene="trfA"
                     /product="TrfA protein"
                     /label=trfA
                     /protein_id="AOF43190.1"
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWLQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMDFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     gene            2892..4040
                     /gene="trfA"
                     /label=trfA
     CDS             complement(4208..4867)
                     /codon_start=1
                     /gene="CmR"
                     /product="chloramphenicol resistance protein"
                     /label=CmR
                     /protein_id="AOF43192.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     gene            complement(4208..4867)
                     /gene="CmR"
                     /label=CmR
     promoter        complement(4868..4970)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     gene            5286..6383
                     /gene="ura3"
                     /label=ura3
     promoter        5286..5501
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             5502..6305
                     /codon_start=1
                     /gene="ura3"
                     /product="URA3"
                     /label=ura3
                     /note="derived from Saccharomyces cerevisiae"
                     /protein_id="AOF43191.1"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      complement(7032..7557)
                     /direction=LEFT
                     /label=oriV
                     /note="oriV"
     rep_origin      7034..7648
                     /label=oriV
                     /note="origin of replication for the bacterial F plasmid"
     rep_origin      7748..7967
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     rep_origin      7748..7964
                     /label=oriS
                     /note="oriS"
     CDS             8058..8813
                     /codon_start=1
                     /gene="repE"
                     /product="RepE protein"
                     /label=repE
                     /protein_id="AOF43188.1"
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     gene            8058..8813
                     /gene="repE"
                     /label=repE
     misc_feature    8816..9066
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             9392..10567
                     /codon_start=1
                     /gene="parA"
                     /product="ParA protein"
                     /label=parA
                     /protein_id="AOF43189.1"
                     /translation="MFRMKLMETLNQCINAGHEMTKAIAIAQFNDDSPEARKITRRWRI
                     GEAADLVGVSSQAIRDAEKAGRLPHPDMEIRGRVEQRVGYTIEQINHMRDVFGTRLRRA
                     EDVFPPVIGVAAHKGGVYKTSVSVHLAQDLALKGLRVLLVEGNDPQGTASMYHGWVPDL
                     HIHAEDTLLPFYLGEKDDVTYAIKPTCWPGLDIIPSCLALHRIETELMGKFDEGKLPTD
                     PHLMLRLAIETVAHDYDVIVIDSAPNLGIGTINVVCAADVLIVPTPAELFDYTSALQFF
                     DMLRDLLKNVDLKGFEPDVRILLTKYSNSNGSQSPWMEEQIRDAWGSMVLKNVVRETDE
                     VGKGQIRMRTVFEQAIDQRSSTGAWRNALSIWEPVCNEIFDRLIKPRWEIR"
     gene            9392..10567
                     /gene="parA"
                     /label=parA
     CDS             10567..11538
                     /codon_start=1
                     /gene="parB"
                     /product="ParB protein"
                     /label=parB
                     /protein_id="AOF43194.1"
                     /translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA
                     ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP
                     AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
                     GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
                     ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
                     APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
     gene            10567..11538
                     /gene="parB"
                     /label=parB
     repeat_region   11611..12084
                     /label=parC
                     /note="parC"
     misc_feature    11611..12084
                     /gene="sopC"
                     /label=sopC
                     /note="centromere-like partitioning region of the bacterial
                     F plasmid"