LOCUS       sequence-407993-        9340 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Broad host-range bacterial expression vector with constitutive Pc 
            promoter. Provides E. coli TorA signal peptide in frame with 
            mNeonGreen (codon optimized for expression in P. fluorescens); for 
            targeting mNeonGreen to the periplasm.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9340)
  AUTHORS   Wilton R, Ahrendt AJ, Shinde S, Sholto-Douglas DJ, Johnson JL, 
            Brennan MB, Kemner KM
  TITLE     A New Suite of Plasmid Vectors for Fluorescence-Based Imaging of 
            Root Colonizing Pseudomonads.
  JOURNAL   Front Plant Sci. 2018 Feb 1;8:2242. doi: 10.3389/fpls.2017.02242. 
            eCollection 2017.
  PUBMED    29449848
REFERENCE   2  (bases 1 to 9340)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Front Plant
            Sci."; date: "2018-02-1"; volume: "8:2242. doi"; pages: " 
            10.3389/fpls.2017.02242. eCollection 2017"
COMMENT     Sequence Label: pSW002-Pc-TorA(sp)-mNeonGreen
FEATURES             Location/Qualifiers
     source          1..9340
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(44..1243)
                     /codon_start=1
                     /gene="tetA"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKPNIPLIVILSTVALDAVGIGLIMPVLPGLLRDLVHSNDVTAHY
                     GILLALYALVQFACAPVLGALSDRFGRRPILLVSLAGATVDYAIMATAPFLWVLYIGRI
                     VAGITGATGAVAGAYIADITDGDERARHFGFMSACFGFGMVAGPVLGGLMGGFSPHAPF
                     FAAAALNGLNFLTGCFLLPESHKGERRPLRREALNPLASFRWARGMTVVAALMAVFFIM
                     QLVGQVPAALWVIFGEDRFHWDATTIGISLAAFGILHSLAQAMITGPVAARLGERRALM
                     LGMIADGTGYILLAFATRGWMAFPIMVLLASGGIGMPALQAMLSRQVDEERQGQLQGSL
                     AALTSLTSIVGPLLFTAIYAASITTWNGWAWIAGAALYLLCLPALRRGLWSGAGQRADR
                     "
     CDS             1349..1999
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline resistance regulatory protein"
                     /label=TetR
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGDD"
     RBS             3003..3025
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3879..3896
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4383..4400)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4517..5062)
                     /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.
                     "
     primer_bind     complement(4597..4616)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     rep_origin      5771..5965
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(6031..7104)
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     CDS             complement(7533..8162)
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
ORIGIN
        1 atagttctag ctagtactag agcggcccct atcgtttcca cgatcagcga tcggctcgtt
       61 gccctgcgcc gctccaaagc ccgcgacgca gcgccggcag gcagagcaag tagagggcag
      121 cgcctgcaat ccatgcccac ccgttccacg ttgttataga agccgcatag atcgccgtga
      181 agaggagggg tccgacgatc gaggtcaggc tggtgagcgc cgccagtgag ccttgcagct
      241 gcccctgacg ttcctcatcc acctgcctgg acaacattgc ttgcagcgcc ggcattccga
      301 tgccacccga agcaagcagg accatgatcg ggaacgccat ccatccccgt gtcgcgaagg
      361 caagcaggat gtagcctgtg ccgtcggcaa tcattccgag catgagtgcc cgcctttcgc
      421 cgagccgggc ggctacaggg ccggtgatca ttgcctgggc gagtgaatgc agaatgccaa
      481 atgcggcaag cgaaatgccg atcgtggtcg cgtcccagtg aaagcgatcc tcgccgaaaa
      541 tgacccaaag cgcggccggc acctgtccga caagttgcat gatgaagaag accgccatca
      601 gggcggcgac gacggtcatg ccccgggccc accggaacga agcgagcggg ttgagagcct
      661 cccggcgtaa cggccggcgt tcgcctttgt gcgactccgg caaaaggaaa cagcccgtca
      721 ggaaattgag gccgttcaag gctgccgcgg cgaagaacgg agcgtggggg gagaaaccgc
      781 ccatcagccc accgagcaca ggtcccgcga ccatcccgaa cccgaaacag gcgctcatga
      841 agccgaagtg ccgcgcgcgc tcatcgccat cagtgatatc ggcaatataa gcgccggcta
      901 ccgccccagt cgccccggtg atgccggcca cgatccgccc gatatagaga acccaaagga
      961 aaggcgctgt cgccatgatg gcgtagtcga cagtggcgcc ggccagcgag acgagcaaga
     1021 ttggccgccg cccgaaacga tccgacagcg cgcccagcac aggtgcgcag gcaaattgca
     1081 ccaacgcata cagcgccagc agaatgccat agtgggcggt gacgtcgttc gagtgaacca
     1141 gatcgcgcag gaggcccggc agcaccggca taatcaggcc gatgccgaca gcgtcgagcg
     1201 cgacagtgct cagaattacg atcaggggta tgttgggttt cacgtctggc ctccggacca
     1261 gcctccgctg gtccgattga acgcgcggat tctttatcac tgataagttg gtggacatat
     1321 tatgtttatc agtgataaag tgtcaagcat gacaaagttg cagccgaata cagtgatccg
     1381 tgccgccctg gacctgttga acgaggtcgg cgtagacggt ctgacgacac gcaaactggc
     1441 ggaacggttg ggggttcagc agccggcgct ttactggcac ttcaggaaca agcgggcgct
     1501 gctcgacgca ctggccgaag ccatgctggc ggagaatcat acgcattcgg tgccgagagc
     1561 cgacgacgac tggcgctcat ttctgatcgg gaatgcccgc agcttcaggc aggcgctgct
     1621 cgcctaccgc gatggcgcgc gcatccatgc cggcacgcga ccgggcgcac cgcagatgga
     1681 aacggccgac gcgcagcttc gcttcctctg cgaggcgggt ttttcggccg gggacgccgt
     1741 caatgcgctg atgacaatca gctacttcac tgttggggcc gtgcttgagg agcaggccgg
     1801 cgacagcgat gccggcgagc gcggcggcac cgttgaacag gctccgctct cgccgctgtt
     1861 gcgggccgcg atagacgcct tcgacgaagc cggtccggac gcagcgttcg agcagggact
     1921 cgcggtgatt gtcgatggat tggcgaaaag gaggctcgtt gtcaggaacg ttgaaggacc
     1981 gagaaagggt gacgattgat caggaccgct gccggagcgc aacccactca ctacagcaga
     2041 gccatgtaga caacatcccc tccccctttc caccgcgtca gacgcccgta gcagcccgct
     2101 acgggctttt tcatgccctg ccctagcgtc caagcctcac ggccgcgctc ggcctctctg
     2161 gcggccttct ggcgctgagg tctgcctcgt gaagaaggtg ttgctgactc ataccaggcc
     2221 tgaatcgccc catcatccag ccagaaagtg agggagccac ggttgatgag agctttgttg
     2281 taggtggacc agttggtgat tttgaacttt tgctttgcca cggaacggtc tgcgttgtcg
     2341 ggaagatgcg tgatctgatc cttcaactca gcaaaagttc gatttattca acaaagccgc
     2401 cgtcccgtca agtcagcgta atgctctgcc agtgttacaa ccaattaacc aattctgatt
     2461 agaaaaactc atcgagcatc aaatgaaact gcaatttatt catatcagga ttatcaatac
     2521 catatttttg aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg cagttccata
     2581 ggatggcaag atcctggtat cggtctgcga ttccgactcg tccaacatca atacaaccta
     2641 ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa atcaccatga gtgacgactg
     2701 aatccggtga gaatggcaaa agctggccgc cacggcgata tcggatccat atgacgtcga
     2761 cgcgttgccg atacaagaac aacagcgcgt tgagcgcctg ccggtgggtg gccggcgcca
     2821 cttgcttctc ggtggcgagc atggtcagaa aaccctcgac ttcagcttgc cccatttcgc
     2881 gcggatgtcg aaacccacca aggctgcggg ccgtccacaa cacaaatgcc ttggcccagt
     2941 agacataagc cttctcggtc tgtaggctgt aatgcaggta gcgaacccgt tcaagaaata
     3001 attttgttta actttaagaa ggagatatac atatgaacaa taacgatctc tttcaggcat
     3061 cacgtcggcg ttttctggca caactcggcg gcttaaccgt cgccgggatg ctggggccgt
     3121 cattgttaac gccgcgacgt gcgactgcgg cgcaagcggc gaattcgaaa ggcgaagaag
     3181 ataacatggc cagtttaccg gcaactcatg aattacatat ttttggaagt attaacggcg
     3241 ttgactttga tatggttgga cagggcaccg gcaacccgaa cgatggttat gaagaattaa
     3301 acctgaaatc tacaaaaggc gacctgcaat tctctccctg gattcttgtt ccacacattg
     3361 gttatggctt ccaccaatac ttaccctatc ctgatggcat gtcccctttc caagccgcga
     3421 tggtggatgg ctctggttac caagtacacc gtaccatgca attcgaagat ggagcatcac
     3481 tcacagtaaa ctaccgctat acatatgaag gttcccacat taaaggcgaa gcacaagtta
     3541 aaggtactgg attccccgct gatggaccag ttatgactaa ctcattgacc gcagcagact
     3601 ggtgtcgctc aaaaaaaacc tatccgaacg ataaaacgat tattagcaca tttaaatgga
     3661 gttatacgac cggaaatggc aaacgttacc gcagcacagc acgcacaacc tatacatttg
     3721 caaaaccaat ggccgcgaat tatttgaaaa accaaccgat gtatgtcttt cgtaaaacag
     3781 aattaaaaca cagcaaaaca gaattaaatt ttaaagaatg gcaaaaagca tttaccgacg
     3841 ttatgggaat ggatgaatta tataaataag cggccgcaca tcatcatcac catcactaag
     3901 ctttaagcat ctcgaggcct cggactagcg gatcagtgag ggtttgcaac tgcgggtcaa
     3961 ggatctggat ttcgatcacg gcacgatcat cgtgcgggag ggcaagggct ccaaggatcg
     4021 ggccttgatg ttacccgaga gcttggcacc cagcctgcgc gagcagggga attgatccgg
     4081 tggatgacct tttgaatgac ctttaataga ttatattact aattaattgg ggaccctaga
     4141 ggtccccttt tttattttaa aaattttttc acaaaacggt ttacaagcat aaagctcctc
     4201 ggactacctg gtatgagtca gcaacacctt cttcacgagg cagacctcag cgctagcgga
     4261 gtgtatactg gcttactatg ttggcactga tgagggtgtc agtgaagtgc ttcatgtggc
     4321 aggagaaaaa aggctgcacc ggtgcgtcag cagaatatgt gatacaggat atattccgct
     4381 tcctcgctca ctgactcgct acgctcggtc gttcgactgc ggcgagcgga aatggcttac
     4441 gaacggggcg gagatttcct ggaagatgcc aggaagatac ttaacaggga agtgagaggg
     4501 ccgcggcaaa gccgtttttc cataggctcc gcccccctga caagcatcac gaaatctgac
     4561 gctcaaatca gtggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     4621 gcggctccct cgtgcgctct cctgttcctg cctttcggtt taccggtgtc attccgctgt
     4681 tatggccgcg tttgtctcat tccacgcctg acactcagtt ccgggtaggc agttcgctcc
     4741 aagctggact gtatgcacga accccccgtt cagtccgacc gctgcgcctt atccggtaac
     4801 tatcgtcttg agtccaaccc ggaaagacat gcaaaagcac cactggcagc agccactggt
     4861 aattgattta gaggagttag tcttgaagtc atgcgccggt taaggctaaa ctgaaaggac
     4921 aagttttggt gactgcgctc ctccaagcca gttacctcgg ttcaaagagt tggtagctca
     4981 gagaaccttc gaaaaaccgc cctgcaaggc ggttttttcg ttttcagagc aagagattac
     5041 gcgcagacca aaacgatctc aagaagatca tcttattaag gggtctgacg ctcagtggaa
     5101 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     5161 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     5221 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     5281 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     5341 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     5401 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     5461 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     5521 gcgcaacgtt gttgccattg cctggtatga gtcagcaaca ccttcttcac gaggcagacc
     5581 tcagcgccgg cggtcgagtg gcgacggcgc ggcttgtccg cgccctggta gattgcctgg
     5641 ccgtaggcca gccatttttg agcggccatc ggccgcgata ggccgacgcg aagcggcggg
     5701 gcgtagggag cgcagcgacc gaagggtagg cgctttttgc agctcttcgg ctgtgcgctg
     5761 gccagacagt tatgcacagg ccaggcgggt tttaagagtt ttaataagtt ttaaagagtt
     5821 ttaggcggaa aaatcgcctt ttttctcttt tatatcagtc acttacatgt gtgaccggtt
     5881 cccaatgtac ggctttgggt tcccaatgta cgggttccgg ttcccaatgt acggctttgg
     5941 gttcccaatg tacgtgctat ccacaggaaa gagacctttt cgaccttttt cccctgctag
     6001 ggcaatttgc cctagcatct gctccgtaca ttaggaaccg gcggatgctt cgccctcgat
     6061 caggttgcgg tagcgcatga ctaggatcgg gccagcctgc cccgcctcct ccttcaaatc
     6121 gtactccggc aggtcatttg acccgatcag cttgcgcacg gtgaaacaga acttcttgaa
     6181 ctctccggcg ctgccactgc gttcgtagat cgtcttgaac aaccatctgg cttctgcctt
     6241 gcctgcggcg cggcgtgcca ggcggtagag aaaacggccg atgccgggat cgatcaaaaa
     6301 gtaatcgggg tgaaccgtca gcacgtccgg gttcttgcct tctgtgatct cgcggtacat
     6361 ccaatcagct agctcgatct cgatgtactc cggccgcccg gtttcgctct ttacgatctt
     6421 gtagcggcta atcaaggctt caccctcgga taccgtcacc aggcggccgt tcttggcctt
     6481 cttcgtacgc tgcatggcaa cgtgcgtggt gtttaaccga atgcaggttt ctaccaggtc
     6541 gtctttctgc tttccgccat cggctcgccg gcagaacttg agtacgtccg caacgtgtgg
     6601 acggaacacg cggccgggct tgtctccctt cccttcccgg tatcggttca tggattcggt
     6661 tagatgggaa accgccatca gtaccaggtc gtaatcccac acactggcca tgccggccgg
     6721 ccctgcggaa acctctacgt gcccgtctgg aagctcgtag cggatcacct cgccagctcg
     6781 tcggtcacgc ttcgacagac ggaaaacggc cacgtccatg atgctgcgac tatcgcgggt
     6841 gcccacgtca tagagcatcg gaacgaaaaa atctggttgc tcgtcgccct tgggcggctt
     6901 cctaatcgac ggcgcaccgg ctgccggcgg ttgccgggat tctttgcgga ttcgatcagc
     6961 ggctgcttgc cacgattcac cggggcgtgc ttctgcctcg atgcgttgcc gctgggcggc
     7021 ctgcgcggcc ttcaacttct ccaccaggtc atcacccagc gccgcgccga tttgtaccgg
     7081 gccggatggt ttgcgaccgc tcacgccgat tcctcgggct tgggggttcc agtgccattg
     7141 cagggccggc agacaaccca gccgcttacg cctggccaac cgcccgttcc tccacacatg
     7201 gggcattcca cggcgtcggt gcctggttgt tcttgatttt ccatgccgcc tcctttagcc
     7261 gctaaaattc atctactcat ttattcattt gctcatttac tctggtagct gcgcgatgta
     7321 ttcagatagc agctcggtaa tggtcttgcc ttggcgtacc gcgtacatct tcagcttggt
     7381 gtgatcctcc gccggcaact gaaagttgac ccgcttcatg gctggcgtgt ctgccaggct
     7441 ggccaacgtt gcagccttgc tgctgcgtgc gctcggacgg ccggcactta gcgtgtttgt
     7501 gcttttgctc attttctctt tacctcatta actcaaatga gttttgattt aatttcagcg
     7561 gccagcgcct ggacctcgcg ggcagcgtcg ccctcgggtt ctgattcaag aacggttgtg
     7621 ccggcggcgg cagtgcctgg gtagctcacg cgctgcgtga tacgggactc aagaatgggc
     7681 agctcgtacc cggccagcgc ctcggcaacc tcaccgccga tgcgcgtgcc tttgatcgcc
     7741 cgcgacacga caaaggccgc ttgtagcctt ccatccgtga cctcaatgcg ctgcttaacc
     7801 agctccacca ggtcggcggt ggcccatatg tcgtaagggc ttggctgcac cggaatcagc
     7861 acgaagtcgg ctgccttgat cgcggacaca gccaagtccg ccgcctgggg cgctccgtcg
     7921 atcactacga agtcgcgccg gccgatggcc ttcacgtcgc ggtcaatcgt cgggcggtcg
     7981 atgccgacaa cggttagcgg ttgatcttcc cgcacggccg cccaatcgcg ggcactgccc
     8041 tggggatcgg aatcgactaa cagaacatcg gccccggcga gttgcagggc gcgggctaga
     8101 tgggttgcga tggtcgtctt gcctgacccg cctttctggt taagtacagc gataaccttc
     8161 atgcgttccc cttgcgtatt tgtttattta ctcatcgcat catatacgca gcgaccgcat
     8221 gacgcaagct gttttactca aatacacatc acctttttag acggcggcgc tcggtttctt
     8281 cagcggccaa gctggccggc caggccgcca gcttggcatc agacaaaccg gccaggattt
     8341 catgcagccg cacggttgag acgtgcgcgg gcggctcgaa cacgtacccg gccgcgatca
     8401 tctccgcctc gatctcttcg gtaatgaaaa acggttcgtc ctggccgtcc tggtgcggtt
     8461 tcatgcttgt tcctcttggc gttcattctc ggcggctgcc agggcgtcgg cctcggtcaa
     8521 tgcgtcctca cggaaggcac cgcgccgcct ggcctcggtg ggcgtcactt cctcgctgcg
     8581 ctcaagtgcg cggtacaggg tcgagcgatg cacgccaagc agtgcagccg cctctttcac
     8641 ggtgcggcct tcctggtcga tcagctcgcg ggcgtgcgcg atctgtgccg gggtgagggt
     8701 agggcggggg ccaaacttca cgcctcgggc cttggcggcc tcgcgcccgc tccgggtgcg
     8761 gtcgatgatt agggaacgct cgaactcggc aatgccggcg aacacggtca acaccatgcg
     8821 gccggccggc gtggtggtgt cggcccacgg ctctgccagg ctacgcaggc ccgcgccggc
     8881 ctcctggatg cgctcggcaa tgtccagtag gtcgcgggtg ctgcgggcca ggcggtctag
     8941 cctggtcact gtcacaacgt cgccagggcg taggtggtca agcatcctgg ccagctccgg
     9001 gcggtcgcgc ctggtgccgg tgatcttctc ggaaaacagc ttggtgcagc cggccgcgtg
     9061 cagttcggcc cgttggttgg tcaagtcctg gtcgtcggtg ctgacgcggg catagcccag
     9121 caggccagcg gcggcgctct tgttcatggc gtaatgtctc cggttctagt cgcaagtatt
     9181 ctactttatg cgactaaaac acgcgacaag aaaacgccag gaaaagggca gggcggcagc
     9241 ctgtcgcgta acttaggact tgtgcgacat gtcgttttca gaagacggct gcactgaacg
     9301 tcagaagccg actgcactat agcagcggag gggttggatc
//
