LOCUS       Exported                5664 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pDS1028, complete sequence.
ACCESSION   KT991389
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5664)
  AUTHORS   Monson RE, Drew AM, Smith D, Matilla M, Roberts K, Richardson E, 
            Williamson N, Ramsay J, Welch M, Salmond GPC.
  TITLE     A plasmid-transposon hybrid mutagenesis system effective in a broad 
            range of Enterobacteria
  JOURNAL   Front Microbiol (2016) In press
REFERENCE   2  (bases 1 to 5664)
  AUTHORS   Monson RE, Smith D, Matilla M, Roberts K, Richardson E, Drew A, 
            Williamson N, Ramsay J, Welch M, Salmond GPC.
  TITLE     Direct Submission
  JOURNAL   Submitted (03-NOV-2015) Biochemistry, University of Cambridge, 
            Tennis Court Road, Cambridge CB2 1QW, United Kingdom
REFERENCE   3  (bases 1 to 5664)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..5664
                     /organism="Cloning vector pDS1028"
                     /mol_type="other DNA"
                     /db_xref="taxon:1767045"
     CDS             complement(129..1319)
                     /codon_start=1
                     /gene="tetA"
                     /product="TetA"
                     /label=tetA
                     /note="tetracycline resistance"
                     /protein_id="ALS31080.1"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     gene            complement(129..1319)
                     /gene="tetA"
                     /label=tetA
     promoter        complement(1367..1395)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             1698..3203
                     /codon_start=1
                     /gene="tnp"
                     /product="transposase"
                     /label=tnp
                     /protein_id="ALS31078.1"
                     /translation="MSTNLSVIKNPRVQSDQRRLVRRPHMITSALHRAADWAKSVFSSA
                     ALGDPRRTARLVNVAAQLAKYSGKSITISSEGSKAAQEGAYRFIRNPNVSAEAIRKAGA
                     MQTVKLAQEFPELLAIEDTTSLSYRHQVAEELGKLGSIQDKSRGWWVHSVLLLEATTFR
                     TVGLLHQEWWMRPDDPADADEKESGKWLAAAATSRLRMGSMMSNVIAVCDREADIHAYL
                     QDKLAHNERFVVRSKHPRKDVESGLYLYDHLKNQPELGGYQISIPQKGVVDKRGKRKNR
                     PARKASLSLRSGRITLKQGNITLNAVLAEEINPPKGETPLKWLLLTSEPVESLAQALRV
                     IDIYTHRWRIEEFHKAWKTGAGAERQRMEEPDNLERMVSILSFVAVRLLQLRESFTPPQ
                     ALRAQGLLKEAEHVESQSAETVLTPDECQLLGYLDKGKRKRKEKAGSLQWAYMAIARLG
                     GFMDSKRTGIASWGALWEGWEALQSKLDGFLAAKDLMAQGIKI"
     gene            1698..3203
                     /gene="tnp"
                     /label=tnp
     misc_feature    3212..3230
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase 
                     recognition (Reznikoff et al., 2004)"
     rep_origin      3269..3657
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     promoter        3974..4076
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             4077..4736
                     /codon_start=1
                     /gene="cat"
                     /product="Cat"
                     /label=cat
                     /note="chloramphenicol resistance"
                     /protein_id="ALS31079.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     gene            4077..4736
                     /gene="cat"
                     /label=cat
     misc_feature    5121..5139
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase 
                     recognition (Reznikoff et al., 2004)"
     oriT            5378..5487
                     /note="incP origin of transfer"