pME6031 载体 (V004776#)

载体基本信息
载体名称 pME6031 载体抗性 Tetracycline
载体长度 8310 bp 载体类型 Shuttle vector
载体来源 Heeb S, Itoh Y, Nishijyo T, Schnider U, Keel C, Wade J, Walsh U, O'Gara F, Haas D.

pME6031 载体载体图谱

pME60318310 bp400800120016002000240028003200360040004400480052005600600064006800720076008000pVS1 resolvaseStaA; similar to IncC2hypothetical proteinKorB boxRepApVS1 origin of replicationp15A origin of replicationpart of terminator absent in pME6030multiple cloning siteTetRTetA

pME6031 载体载体序列

LOCUS       Exported                8310 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pME6031, complete sequence.
ACCESSION   AF118811
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8310)
  AUTHORS   Heeb S, Itoh Y, Nishijyo T, Schnider U, Keel C, Wade J, Walsh U, 
            O'Gara F, Haas D.
  TITLE     Small, stable shuttle vectors based on the minimal pVS1 replicon for
            use in gram-negative, plant-associated bacteria
  JOURNAL   Mol. Plant Microbe Interact. 13 (2), 232-237 (2000)
  PUBMED    10659714
REFERENCE   2  (bases 1 to 8310)
  AUTHORS   Heeb S.
  TITLE     Direct Submission
  JOURNAL   Submitted (30-DEC-1998) Laboratoire de Biologie Microbienne, 
            University of Lausanne, Batiment de Biologie, Lausanne CH-1015, 
            Switzerland
REFERENCE   3  (bases 1 to 8310)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8310
                     /organism="Shuttle vector pME6031"
                     /host="Pseudomonas pVS1; Escherichia p15A replicons"
                     /mol_type="other DNA"
                     /label=derived from plasmids pACYC177, pVK100, and pME290
                     /note="derived from plasmids pACYC177, pVK100, and pME290"
                     /db_xref="taxon:89145"
     CDS             195..881
                     /codon_start=1
                     /product="pVS1 resolvase"
                     /label=pVS1 resolvase
                     /note="ORF1; not required for stable maintenance"
                     /protein_id="AAD19683.1"
                     /translation="MNKSAAAGLLGYARVSTDDQDLTNQRAELHAAGCTKLFSEKITGT
                     RRDRPELARMLDHLRPGDVVTVTRLDRLARSTRDLLDIAERIQEAGAGLRSLAEPWADT
                     TTPAGRMVLTVFAGIAEFERSLIIDRTRSGREAAKARGVKFGPRPTLTPAQIAHARELI
                     DQEGRTVKEAAALLGVHRSTLYRALERSEEVTPTEARRRGAFREDALTEADALAAAENE
                     RQEEQA"
     CDS             878..1093
                     /codon_start=1
                     /product="hypothetical protein"
                     /label=hypothetical protein
                     /note="ORF2; not required for stable maintenance"
                     /protein_id="AAD19684.1"
                     /translation="MKPHQDGQDEPFFITEEIEAEMIAAGYVFEPPAHVSTVRLHEILA
                     GLSDAKLAAWPASLAAEETERRRLKR"
     CDS             1180..1809
                     /codon_start=1
                     /product="pVS1 partitioning protein"
                     /label=pVS1 partitioning protein
                     /note="StaA; similar to IncC2"
                     /protein_id="AAD19685.1"
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             1830..2045
                     /codon_start=1
                     /product="hypothetical protein"
                     /label=hypothetical protein
                     /note="ORF3; not required for stable maintenance"
                     /protein_id="AAD19686.1"
                     /translation="MSKSTNTLSAGRPSARSSKAATLASLADTPAMKRVNFQLPAEDHT
                     KLKMYAVRQGKTITELLSEYIAQLPE"
     misc_feature    2075..2088
                     /label=KorB box
                     /note="KorB box"
     CDS             2238..3311
                     /codon_start=1
                     /product="pVS1 replication protein"
                     /label=pVS1 replication protein
                     /note="RepA"
                     /protein_id="AAD19687.1"
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     rep_origin      3377..3571
                     /label=pVS1 origin of replication
                     /note="pVS1 origin of replication"
     misc_feature    3377..3384
                     /label=DnaA box
                     /note="DnaA box"
     repeat_region   3388..3471
                     /label=21 bp repeats
                     /note="21 bp repeats"
     misc_feature    3472..3563
                     /label=AT-rich region
                     /note="AT-rich region"
     misc_feature    3564..3571
                     /label=DnaA box
                     /note="DnaA box"
     rep_origin      4264..5093
                     /label=p15A origin of replication
                     /note="p15A origin of replication"
     rep_origin      4281..4826
                     /direction=RIGHT
                     /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."
     misc_feature    5068..5089
                     /label=p15A origin of transfer
                     /note="p15A origin of transfer"
     misc_difference 5136..5414
                     /label=part of terminator absent in pME6030
                     /note="part of terminator absent in pME6030"
     regulatory      5148..5414
                     /regulatory_class="terminator"
                     /note="T4 transcription terminator"
     misc_feature    5416..5526
                     /label=multiple cloning site
                     /note="multiple cloning site"
     CDS             complement(6315..6992)
                     /codon_start=1
                     /product="RK2 tetracycline repressor protein"
                     /label=RK2 tetracycline repressor protein
                     /note="TetR"
                     /protein_id="AAD19689.1"
                     /translation="MFISDKVSSMTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGV
                     QQPALYWHFRNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRD
                     GARIHAGTRPGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSD
                     AGERGGTVEQAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRK
                     GDD"
     CDS             complement(6315..6965)
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline resistance regulatory protein"
                     /label=TetR
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGDD"
     CDS             6996..8270
                     /codon_start=1
                     /product="RK2 tetracycline resistance protein"
                     /label=RK2 tetracycline resistance protein
                     /note="TetA"
                     /protein_id="AAD19688.1"
                     /translation="MSTNLSVIKNPRVQSDQRRLVRRPDVKPNIPLIVILSTVALDAVG
                     IGLIMPVLPGLLRDLVHSNDVTAHYGILLALYALVQFACAPVLGALSDRFGRRPILLVS
                     LAGATVDYAIMATAPFLWVLYIGRIVAGITGATGAVAGAYIADITDGDERARHFGFMSA
                     CFGFGMVAGPVLGGLMGGFSPHAPFFAAAALNGLNFLTGCFLLPESHKGERRPLRREAL
                     NPLASFRWARGMTVVAALMAVFFIMQLVGQVPAALWVIFGEDRFHWDATTIGISLAAFG
                     ILHSLAQAMITGPVAARLGERRALMLGMIADGTGYILLAFATRGWMAFPIMVLLASGGI
                     GMPALQAMLSRQVDEERQGQLQGSLAALTSLTSIVGPLLFTAIYAASITTWNGWAWIAG
                     AALYLLCLPALRRGLWSGAGQRADR"
     CDS             7071..8270
                     /codon_start=1
                     /gene="tetA"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKPNIPLIVILSTVALDAVGIGLIMPVLPGLLRDLVHSNDVTAHY
                     GILLALYALVQFACAPVLGALSDRFGRRPILLVSLAGATVDYAIMATAPFLWVLYIGRI
                     VAGITGATGAVAGAYIADITDGDERARHFGFMSACFGFGMVAGPVLGGLMGGFSPHAPF
                     FAAAALNGLNFLTGCFLLPESHKGERRPLRREALNPLASFRWARGMTVVAALMAVFFIM
                     QLVGQVPAALWVIFGEDRFHWDATTIGISLAAFGILHSLAQAMITGPVAARLGERRALM
                     LGMIADGTGYILLAFATRGWMAFPIMVLLASGGIGMPALQAMLSRQVDEERQGQLQGSL
                     AALTSLTSIVGPLLFTAIYAASITTWNGWAWIAGAALYLLCLPALRRGLWSGAGQRADR
                     "

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