LOCUS       Exported                8612 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pTSara*, complete sequence.
ACCESSION   KX986162
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8612)
  AUTHORS   Boehm CR, Grant PK, Haseloff J.
  TITLE     Programmed emergence of a domain of gene expression in a bacterial 
            population
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8612)
  AUTHORS   Boehm CR.
  TITLE     Direct Submission
  JOURNAL   Submitted (12-OCT-2016) Plant Sciences, University of Cambridge, 
            Downing Street, Cambridge, Cambridgeshire CB2 3EA, United Kingdom
REFERENCE   3  (bases 1 to 8612)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..8612
                     /organism="Cloning vector pTSara*"
                     /mol_type="other DNA"
                     /db_xref="taxon:2049422"
     CDS             complement(96..974)
                     /codon_start=1
                     /product="araC"
                     /label=araC
                     /protein_id="ATP60601.1"
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     regulatory      complement(976..1025)
                     /regulatory_class="promoter"
                     /note="araC promoter"
     regulatory      complement(1158..1252)
                     /regulatory_class="terminator"
                     /note="BBa_B0014"
     terminator      1218..1245
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(1298..1837)
                     /codon_start=1
                     /product="T7RNAP"
                     /label=T7RNAP
                     /protein_id="ATP60602.1"
                     /translation="MNTINIAKNDFSDIELAAIPFNTLADHYGERLAREQLALEHESYE
                     MGEARFRKMFERQLKAGEVADNAAAKPLITTLLPKMIARINDWFEEVKAKRGKRPTAFQ
                     FLQEIKPEAVAYITIKTTLACLTSADNTTVQAVASAIGRAIEDEARFGRIRDLEAKHFK
                     KNVEEQLNKRVGHVYK"
     regulatory      complement(1838..1872)
                     /regulatory_class="ribosome_binding_site"
                     /note="N-terminal 500 RU, from Shis et al. 2013"
     regulatory      complement(1879..2009)
                     /regulatory_class="promoter"
                     /note="pBAD"
     regulatory      2307..2437
                     /regulatory_class="promoter"
                     /note="pBAD"
     regulatory      2444..2474
                     /regulatory_class="ribosome_binding_site"
                     /note="C-terminal 500 RU, from Shis et al. 2013"
     CDS             2475..4592
                     /codon_start=1
                     /product="T7RNAP"
                     /label=T7RNAP
                     /protein_id="ATP60603.1"
                     /translation="MKAFMQVVEADMLSKGLLGGEAWSSWHKEDSIHVGVRCIEMLIES
                     TGMVSLHRQNAGVVGQDSETIELAPEYAEAIATRAGALAGISPMFQPCVVPPKPWTGIT
                     GGGYWANGRRPLALVRTHSKKALMRYEDVYMPEVYKAINIAQNTAWKINKKVLAVANVI
                     TKWKHCPVEDIPAIEREELPMKPEDIDMNPEALTAWKRAAAAVYRKDKARKSRRISLEF
                     MLEQANKFANHKAIWFPYNMDWRGRVYAVSMFNPQGNDMTKGLLTLAKGKPIGKEGYYW
                     LKIHGANCAGVDKVPFPERIKFIEENHENIMACAKSPLENTWWAEQDSPFCFLAFCFEY
                     AGVQHHGLSYNCSLPLAFDGSCSGIQHFSAMLRDEVGGRAVNLLPSETVQDIYGIVAKK
                     VNEILQADAINGTDNEVVTVTDENTGEISEKVKLGTKALAGQWLAYGVTRSVTKSSVMT
                     LAYGSKEFGFRQQVLEDTIQPAIDSGKGLMFTQPNQAAGYMAKLIWESVSVTVVAAVEA
                     MNWLKSAAKLLAAEVKDKKTGEILRKRCAVHWVTPDGFPVWQEYKKPIQTRLNLMFLGQ
                     FRLQPTINTNKDSEIDAHKQESGIAPNFVHSQDGSHLRKTVVWAHEKYGIESFALIHDS
                     FGTIPADAANLFKAVRETMVDTYESCDVLADFYDQFADQLHESQLDKMPALPAKGNLNL
                     RDILESDFAFA"
     regulatory      4705..4862
                     /regulatory_class="terminator"
                     /note="rrnb terminator"
     terminator      4825..4911
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5003..5030
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        5049..5140
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5594..6049
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             complement(6604..7263)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     promoter        complement(7264..7366)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     rep_origin      complement(7892..8437)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin."