LOCUS       sequence-407997-        9214 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Broad host-range bacterial expression vector with constitutive Pc 
            promoter; for tagging bacteria with mNeonGreen (codon optimized for 
            P. fluorescens).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9214)
  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 9214)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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: pSW003-Pc-mNeonGreen
FEATURES             Location/Qualifiers
     source          1..9214
                     /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             3753..3770
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4257..4274)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4391..4936)
                     /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(4471..4490)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     rep_origin      5645..5839
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(5905..6978)
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELVDWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     CDS             complement(7407..8036)
                     /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 atatgaattc gaaaggcgaa gaagataaca
     3061 tggccagttt accggcaact catgaattac atatttttgg aagtattaac ggcgttgact
     3121 ttgatatggt tggacagggc accggcaacc cgaacgatgg ttatgaagaa ttaaacctga
     3181 aatctacaaa aggcgacctg caattctctc cctggattct tgttccacac attggttatg
     3241 gcttccacca atacttaccc tatcctgatg gcatgtcccc tttccaagcc gcgatggtgg
     3301 atggctctgg ttaccaagta caccgtacca tgcaattcga agatggagca tcactcacag
     3361 taaactaccg ctatacatat gaaggttccc acattaaagg cgaagcacaa gttaaaggta
     3421 ctggattccc cgctgatgga ccagttatga ctaactcatt gaccgcagca gactggtgtc
     3481 gctcaaaaaa aacctatccg aacgataaaa cgattattag cacatttaaa tggagttata
     3541 cgaccggaaa tggcaaacgt taccgcagca cagcacgcac aacctataca tttgcaaaac
     3601 caatggccgc gaattatttg aaaaaccaac cgatgtatgt ctttcgtaaa acagaattaa
     3661 aacacagcaa aacagaatta aattttaaag aatggcaaaa agcatttacc gacgttatgg
     3721 gaatggatga attatataaa taagcggccg cacatcatca tcaccatcac taagctttaa
     3781 gcatctcgag gcctcggact agcggatcag tgagggtttg caactgcggg tcaaggatct
     3841 ggatttcgat cacggcacga tcatcgtgcg ggagggcaag ggctccaagg atcgggcctt
     3901 gatgttaccc gagagcttgg cacccagcct gcgcgagcag gggaattgat ccggtggatg
     3961 accttttgaa tgacctttaa tagattatat tactaattaa ttggggaccc tagaggtccc
     4021 cttttttatt ttaaaaattt tttcacaaaa cggtttacaa gcataaagct cctcggacta
     4081 cctggtatga gtcagcaaca ccttcttcac gaggcagacc tcagcgctag cggagtgtat
     4141 actggcttac tatgttggca ctgatgaggg tgtcagtgaa gtgcttcatg tggcaggaga
     4201 aaaaaggctg caccggtgcg tcagcagaat atgtgataca ggatatattc cgcttcctcg
     4261 ctcactgact cgctacgctc ggtcgttcga ctgcggcgag cggaaatggc ttacgaacgg
     4321 ggcggagatt tcctggaaga tgccaggaag atacttaaca gggaagtgag agggccgcgg
     4381 caaagccgtt tttccatagg ctccgccccc ctgacaagca tcacgaaatc tgacgctcaa
     4441 atcagtggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggcggct
     4501 ccctcgtgcg ctctcctgtt cctgcctttc ggtttaccgg tgtcattccg ctgttatggc
     4561 cgcgtttgtc tcattccacg cctgacactc agttccgggt aggcagttcg ctccaagctg
     4621 gactgtatgc acgaaccccc cgttcagtcc gaccgctgcg ccttatccgg taactatcgt
     4681 cttgagtcca acccggaaag acatgcaaaa gcaccactgg cagcagccac tggtaattga
     4741 tttagaggag ttagtcttga agtcatgcgc cggttaaggc taaactgaaa ggacaagttt
     4801 tggtgactgc gctcctccaa gccagttacc tcggttcaaa gagttggtag ctcagagaac
     4861 cttcgaaaaa ccgccctgca aggcggtttt ttcgttttca gagcaagaga ttacgcgcag
     4921 accaaaacga tctcaagaag atcatcttat taaggggtct gacgctcagt ggaacgaaaa
     4981 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     5041 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     5101 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     5161 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     5221 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     5281 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     5341 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     5401 cgttgttgcc attgcctggt atgagtcagc aacaccttct tcacgaggca gacctcagcg
     5461 ccggcggtcg agtggcgacg gcgcggcttg tccgcgccct ggtagattgc ctggccgtag
     5521 gccagccatt tttgagcggc catcggccgc gataggccga cgcgaagcgg cggggcgtag
     5581 ggagcgcagc gaccgaaggg taggcgcttt ttgcagctct tcggctgtgc gctggccaga
     5641 cagttatgca caggccaggc gggttttaag agttttaata agttttaaag agttttaggc
     5701 ggaaaaatcg ccttttttct cttttatatc agtcacttac atgtgtgacc ggttcccaat
     5761 gtacggcttt gggttcccaa tgtacgggtt ccggttccca atgtacggct ttgggttccc
     5821 aatgtacgtg ctatccacag gaaagagacc ttttcgacct ttttcccctg ctagggcaat
     5881 ttgccctagc atctgctccg tacattagga accggcggat gcttcgccct cgatcaggtt
     5941 gcggtagcgc atgactagga tcgggccagc ctgccccgcc tcctccttca aatcgtactc
     6001 cggcaggtca tttgacccga tcagcttgcg cacggtgaaa cagaacttct tgaactctcc
     6061 ggcgctgcca ctgcgttcgt agatcgtctt gaacaaccat ctggcttctg ccttgcctgc
     6121 ggcgcggcgt gccaggcggt agagaaaacg gccgatgccg ggatcgatca aaaagtaatc
     6181 ggggtgaacc gtcagcacgt ccgggttctt gccttctgtg atctcgcggt acatccaatc
     6241 aactagctcg atctcgatgt actccggccg cccggtttcg ctctttacga tcttgtagcg
     6301 gctaatcaag gcttcaccct cggataccgt caccaggcgg ccgttcttgg ccttcttcgt
     6361 acgctgcatg gcaacgtgcg tggtgtttaa ccgaatgcag gtttctacca ggtcgtcttt
     6421 ctgctttccg ccatcggctc gccggcagaa cttgagtacg tccgcaacgt gtggacggaa
     6481 cacgcggccg ggcttgtctc ccttcccttc ccggtatcgg ttcatggatt cggttagatg
     6541 ggaaaccgcc atcagtacca ggtcgtaatc ccacacactg gccatgccgg ccggccctgc
     6601 ggaaacctct acgtgcccgt ctggaagctc gtagcggatc acctcgccag ctcgtcggtc
     6661 acgcttcgac agacggaaaa cggccacgtc catgatgctg cgactatcgc gggtgcccac
     6721 gtcatagagc atcggaacga aaaaatctgg ttgctcgtcg cccttgggcg gcttcctaat
     6781 cgacggcgca ccggctgccg gcggttgccg ggattctttg cggattcgat cagcggctgc
     6841 ttgccacgat tcaccggggc gtgcttctgc ctcgatgcgt tgccgctggg cggcctgcgc
     6901 ggccttcaac ttctccacca ggtcatcacc cagcgccgcg ccgatttgta ccgggccgga
     6961 tggtttgcga ccgctcacgc cgattcctcg ggcttggggg ttccagtgcc attgcagggc
     7021 cggcagacaa cccagccgct tacgcctggc caaccgcccg ttcctccaca catggggcat
     7081 tccacggcgt cggtgcctgg ttgttcttga ttttccatgc cgcctccttt agccgctaaa
     7141 attcatctac tcatttattc atttgctcat ttactctggt agctgcgcga tgtattcaga
     7201 tagcagctcg gtaatggtct tgccttggcg taccgcgtac atcttcagct tggtgtgatc
     7261 ctccgccggc aactgaaagt tgacccgctt catggctggc gtgtctgcca ggctggccaa
     7321 cgttgcagcc ttgctgctgc gtgcgctcgg acggccggca cttagcgtgt ttgtgctttt
     7381 gctcattttc tctttacctc attaactcaa atgagttttg atttaatttc agcggccagc
     7441 gcctggacct cgcgggcagc gtcgccctcg ggttctgatt caagaacggt tgtgccggcg
     7501 gcggcagtgc ctgggtagct cacgcgctgc gtgatacggg actcaagaat gggcagctcg
     7561 tacccggcca gcgcctcggc aacctcaccg ccgatgcgcg tgcctttgat cgcccgcgac
     7621 acgacaaagg ccgcttgtag ccttccatcc gtgacctcaa tgcgctgctt aaccagctcc
     7681 accaggtcgg cggtggccca tatgtcgtaa gggcttggct gcaccggaat cagcacgaag
     7741 tcggctgcct tgatcgcgga cacagccaag tccgccgcct ggggcgctcc gtcgatcact
     7801 acgaagtcgc gccggccgat ggccttcacg tcgcggtcaa tcgtcgggcg gtcgatgccg
     7861 acaacggtta gcggttgatc ttcccgcacg gccgcccaat cgcgggcact gccctgggga
     7921 tcggaatcga ctaacagaac atcggccccg gcgagttgca gggcgcgggc tagatgggtt
     7981 gcgatggtcg tcttgcctga cccgcctttc tggttaagta cagcgataac cttcatgcgt
     8041 tccccttgcg tatttgttta tttactcatc gcatcatata cgcagcgacc gcatgacgca
     8101 agctgtttta ctcaaataca catcaccttt ttagacggcg gcgctcggtt tcttcagcgg
     8161 ccaagctggc cggccaggcc gccagcttgg catcagacaa accggccagg atttcatgca
     8221 gccgcacggt tgagacgtgc gcgggcggct cgaacacgta cccggccgcg atcatctccg
     8281 cctcgatctc ttcggtaatg aaaaacggtt cgtcctggcc gtcctggtgc ggtttcatgc
     8341 ttgttcctct tggcgttcat tctcggcggc tgccagggcg tcggcctcgg tcaatgcgtc
     8401 ctcacggaag gcaccgcgcc gcctggcctc ggtgggcgtc acttcctcgc tgcgctcaag
     8461 tgcgcggtac agggtcgagc gatgcacgcc aagcagtgca gccgcctctt tcacggtgcg
     8521 gccttcctgg tcgatcagct cgcgggcgtg cgcgatctgt gccggggtga gggtagggcg
     8581 ggggccaaac ttcacgcctc gggccttggc ggcctcgcgc ccgctccggg tgcggtcgat
     8641 gattagggaa cgctcgaact cggcaatgcc ggcgaacacg gtcaacacca tgcggccggc
     8701 cggcgtggtg gtgtcggccc acggctctgc caggctacgc aggcccgcgc cggcctcctg
     8761 gatgcgctcg gcaatgtcca gtaggtcgcg ggtgctgcgg gccaggcggt ctagcctggt
     8821 cactgtcaca acgtcgccag ggcgtaggtg gtcaagcatc ctggccagct ccgggcggtc
     8881 gcgcctggtg ccggtgatct tctcggaaaa cagcttggtg cagccggccg cgtgcagttc
     8941 ggcccgttgg ttggtcaagt cctggtcgtc ggtgctgacg cgggcatagc ccagcaggcc
     9001 agcggcggcg ctcttgttca tggcgtaatg tctccggttc tagtcgcaag tattctactt
     9061 tatgcgacta aaacacgcga caagaaaacg ccaggaaaag ggcagggcgg cagcctgtcg
     9121 cgtaacttag gacttgtgcg acatgtcgtt ttcagaagac ggctgcactg aacgtcagaa
     9181 gccgactgca ctatagcagc ggaggggttg gatc
//
