pDS1028 vector (V007925)

Basic Vector Information

Vector Name:
pDS1028
Antibiotic Resistance:
Chloramphenicol
Length:
5664 bp
Type:
Cloning vector
Replication origin:
R6K γ ori
Source/Author:
Monson RE, Drew AM, Smith D, Matilla M, Roberts K, Richardson E, Williamson N, Ramsay J, Welch M, Salmond GPC.

pDS1028 vector Map

pDS10285664 bp60012001800240030003600420048005400TcRtet promoterTn5 transposaseTn5 MER6K gamma oricat promoterCmRTn5 MEoriT

pDS1028 vector Sequence

LOCUS       40924_15180        5664 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pDS1028, complete sequence.
ACCESSION   .
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)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5664)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Front 
            Microbiol (2016) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (03-NOV-2015) Biochemistry, University of Cambridge, Tennis Court 
            Road, Cambridge CB2 1QW, United Kingdom"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..5664
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(132..1319)
                     /codon_start=1
                     /label=TcR
                     /note="tetracycline efflux protein"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     promoter        complement(1367..1395)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
     CDS             1773..3200
                     /codon_start=1
                     /label=Tn5 transposase
                     /note="transposase from the bacterial Tn5 transposon
                     (Reznikoff, 1993)"
                     /translation="MITSALHRAADWAKSVFSSAALGDPRRTARLVNVAAQLAKYSGKS
                     ITISSEGSKAAQEGAYRFIRNPNVSAEAIRKAGAMQTVKLAQEFPELLAIEDTTSLSYR
                     HQVAEELGKLGSIQDKSRGWWVHSVLLLEATTFRTVGLLHQEWWMRPDDPADADEKESG
                     KWLAAAATSRLRMGSMMSNVIAVCDREADIHAYLQDKLAHNERFVVRSKHPRKDVESGL
                     YLYDHLKNQPELGGYQISIPQKGVVDKRGKRKNRPARKASLSLRSGRITLKQGNITLNA
                     VLAEEINPPKGETPLKWLLLTSEPVESLAQALRVIDIYTHRWRIEEFHKAWKTGAGAER
                     QRMEEPDNLERMVSILSFVAVRLLQLRESFTPPQALRAQGLLKEAEHVESQSAETVLTP
                     DECQLLGYLDKGKRKRKEKAGSLQWAYMAIARLGGFMDSKRTGIASWGALWEGWEALQS
                     KLDGFLAAKDLMAQGIKI"
     misc_feature    3212..3230
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase
                     recognition (Reznikoff et al., 2004)"
     rep_origin      complement(3269..3657)
                     /direction=LEFT
                     /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..4733
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     misc_feature    complement(5121..5139)
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase
                     recognition (Reznikoff et al., 2004)"
     oriT            complement(5378..5487)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"

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