LOCUS       sequence-215261-        9094 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Broad host-range bacterial expression vector with constitutive PsbA 
            promoter; for tagging bacteria with E2-Crimson.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9094)
  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 9094)
  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-PpsbA-E2-Crimson
FEATURES             Location/Qualifiers
     source          1..9094
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1179..1808
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             2237..3310
                     /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"
     rep_origin      3376..3570
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     rep_origin      4279..4824
                     /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.
                     "
     primer_bind     4725..4744
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4941..4958
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             complement(5434..6111)
                     /codon_start=1
                     /product="far-red noncytotoxic tetrameric variant of DsRed 
                     fluorescent protein (Strack et al., 2009)"
                     /label=E2-Crimson
                     /note="mammalian codon-optimized"
                     /translation="MNSTENVIKPFMRFKVHMEGSVNGHEFEIEGVGEGKPYEGTQTAK
                     LQVTKGGPLPFAWDILSPQFFYGSKAYIKHPADIPDYLKQSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGTLIYHVKFIGVNFPSDGPVMQKKTLGWEPSTERNYPRDGVLKGENHM
                     ALKLKGGGHYLCEFKSIYMAKKPVKLPGYHYVDYKLDITSHNEDYTVVEQYERAEARHH
                     LFQ"
     primer_bind     complement(5694..5713)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     RBS             6119..6141
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             complement(7096..7746)
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline resistance regulatory protein"
                     /label=TetR
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGDD"
     CDS             7852..9051
                     /codon_start=1
                     /gene="tetA"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKPNIPLIVILSTVALDAVGIGLIMPVLPGLLRDLVHSNDVTAHY
                     GILLALYALVQFACAPVLGALSDRFGRRPILLVSLAGATVDYAIMATAPFLWVLYIGRI
                     VAGITGATGAVAGAYIADITDGDERARHFGFMSACFGFGMVAGPVLGGLMGGFSPHAPF
                     FAAAALNGLNFLTGCFLLPESHKGERRPLRREALNPLASFRWARGMTVVAALMAVFFIM
                     QLVGQVPAALWVIFGEDRFHWDATTIGISLAAFGILHSLAQAMITGPVAARLGERRALM
                     LGMIADGTGYILLAFATRGWMAFPIMVLLASGGIGMPALQAMLSRQVDEERQGQLQGSL
                     AALTSLTSIVGPLLFTAIYAASITTWNGWAWIAGAALYLLCLPALRRGLWSGAGQRADR
                     "
ORIGIN
        1 gatccaaccc ctccgctgct atagtgcagt cggcttctga cgttcagtgc agccgtcttc
       61 tgaaaacgac atgtcgcaca agtcctaagt tacgcgacag gctgccgccc tgcccttttc
      121 ctggcgtttt cttgtcgcgt gttttagtcg cataaagtag aatacttgcg actagaaccg
      181 gagacattac gccatgaaca agagcgccgc cgctggcctg ctgggctatg cccgcgtcag
      241 caccgacgac caggacttga ccaaccaacg ggccgaactg cacgcggccg gctgcaccaa
      301 gctgttttcc gagaagatca ccggcaccag gcgcgaccgc ccggagctgg ccaggatgct
      361 tgaccaccta cgccctggcg acgttgtgac agtgaccagg ctagaccgcc tggcccgcag
      421 cacccgcgac ctactggaca ttgccgagcg catccaggag gccggcgcgg gcctgcgtag
      481 cctggcagag ccgtgggccg acaccaccac gccggccggc cgcatggtgt tgaccgtgtt
      541 cgccggcatt gccgagttcg agcgttccct aatcatcgac cgcacccgga gcgggcgcga
      601 ggccgccaag gcccgaggcg tgaagtttgg cccccgccct accctcaccc cggcacagat
      661 cgcgcacgcc cgcgagctga tcgaccagga aggccgcacc gtgaaagagg cggctgcact
      721 gcttggcgtg catcgctcga ccctgtaccg cgcacttgag cgcagcgagg aagtgacgcc
      781 caccgaggcc aggcggcgcg gtgccttccg tgaggacgca ttgaccgagg ccgacgccct
      841 ggcagccgcc gagaatgaac gccaagagga acaagcatga aaccgcacca ggacggccag
      901 gacgaaccgt ttttcattac cgaagagatc gaggcggaga tgatcgcggc cgggtacgtg
      961 ttcgagccgc ccgcgcacgt ctcaaccgtg cggctgcatg aaatcctggc cggtttgtct
     1021 gatgccaagc tggcggcctg gccggccagc ttggccgctg aagaaaccga gcgccgccgt
     1081 ctaaaaaggt gatgtgtatt tgagtaaaac agcttgcgtc atgcggtcgc tgcgtatatg
     1141 atgcgatgag taaataaaca aatacgcaag gggaacgcat gaaggttatc gctgtactta
     1201 accagaaagg cgggtcaggc aagacgacca tcgcaaccca tctagcccgc gccctgcaac
     1261 tcgccggggc cgatgttctg ttagtcgatt ccgatcccca gggcagtgcc cgcgattggg
     1321 cggccgtgcg ggaagatcaa ccgctaaccg ttgtcggcat cgaccgcccg acgattgacc
     1381 gcgacgtgaa ggccatcggc cggcgcgact tcgtagtgat cgacggagcg ccccaggcgg
     1441 cggacttggc tgtgtccgcg atcaaggcag ccgacttcgt gctgattccg gtgcagccaa
     1501 gcccttacga catatgggcc accgccgacc tggtggagct ggttaagcag cgcattgagg
     1561 tcacggatgg aaggctacaa gcggcctttg tcgtgtcgcg ggcgatcaaa ggcacgcgca
     1621 tcggcggtga ggttgccgag gcgctggccg ggtacgagct gcccattctt gagtcccgta
     1681 tcacgcagcg cgtgagctac ccaggcactg ccgccgccgg cacaaccgtt cttgaatcag
     1741 aacccgaggg cgacgctgcc cgcgaggtcc aggcgctggc cgctgaaatt aaatcaaaac
     1801 tcatttgagt taatgaggta aagagaaaat gagcaaaagc acaaacacgc taagtgccgg
     1861 ccgtccgagc gcacgcagca gcaaggctgc aacgttggcc agcctggcag acacgccagc
     1921 catgaagcgg gtcaactttc agttgccggc ggaggatcac accaagctga agatgtacgc
     1981 ggtacgccaa ggcaagacca ttaccgagct gctatctgaa tacatcgcgc agctaccaga
     2041 gtaaatgagc aaatgaataa atgagtagat gaattttagc ggctaaagga ggcggcatgg
     2101 aaaatcaaga acaaccaggc accgacgccg tggaatgccc catgtgtgga ggaacgggcg
     2161 gttggccagg cgtaagcggc tgggttgtct gccggccctg caatggcact ggaaccccca
     2221 agcccgagga atcggcgtga gcggtcgcaa accatccggc ccggtacaaa tcggcgcggc
     2281 gctgggtgat gacctggtgg agaagttgaa ggccgcgcag gccgcccagc ggcaacgcat
     2341 cgaggcagaa gcacgccccg gtgaatcgtg gcaagcagcc gctgatcgaa tccgcaaaga
     2401 atcccggcaa ccgccggcag ccggtgcgcc gtcgattagg aagccgccca agggcgacga
     2461 gcaaccagat tttttcgttc cgatgctcta tgacgtgggc acccgcgata gtcgcagcat
     2521 catggacgtg gccgttttcc gtctgtcgaa gcgtgaccga cgagctggcg aggtgatccg
     2581 ctacgagctt ccagacgggc acgtagaggt ttccgcaggg ccggccggca tggccagtgt
     2641 gtgggattac gacctggtac tgatggcggt ttcccatcta accgaatcca tgaaccgata
     2701 ccgggaaggg aagggagaca agcccggccg cgtgttccgt ccacacgttg cggacgtact
     2761 caagttctgc cggcgagccg atggcggaaa gcagaaagac gacctggtag aaacctgcat
     2821 tcggttaaac accacgcacg ttgccatgca gcgtacgaag aaggccaaga acggccgcct
     2881 ggtgacggta tccgagggtg aagccttgat tagccgctac aagatcgtaa agagcgaaac
     2941 cgggcggccg gagtacatcg agatcgagct agctgattgg atgtaccgcg agatcacaga
     3001 aggcaagaac ccggacgtgc tgacggttca ccccgattac tttttgatcg atcccggcat
     3061 cggccgtttt ctctaccgcc tggcacgccg cgccgcaggc aaggcagaag ccagatggtt
     3121 gttcaagacg atctacgaac gcagtggcag cgccggagag ttcaagaagt tctgtttcac
     3181 cgtgcgcaag ctgatcgggt caaatgacct gccggagtac gatttgaagg aggaggcggg
     3241 gcaggctggc ccgatcctag tcatgcgcta ccgcaacctg atcgagggcg aagcatccgc
     3301 cggttcctaa tgtacggagc agatgctagg gcaaattgcc ctagcagggg aaaaaggtcg
     3361 aaaaggtctc tttcctgtgg atagcacgta cattgggaac ccaaagccgt acattgggaa
     3421 ccggaacccg tacattggga acccaaagcc gtacattggg aaccggtcac acatgtaagt
     3481 gactgatata aaagagaaaa aaggcgattt ttccgcctaa aactctttaa aacttattaa
     3541 aactcttaaa acccgcctgg cctgtgcata actgtctggc cagcgcacag ccgaagagct
     3601 gcaaaaagcg cctacccttc ggtcgctgcg ctccctacgc cccgccgctt cgcgtcggcc
     3661 tatcgcggcc gatggccgct caaaaatggc tggcctacgg ccaggcaatc taccagggcg
     3721 cggacaagcc gcgccgtcgc cactcgaccg ccggcgctga ggtctgcctc gtgaagaagg
     3781 tgttgctgac tcataccagg caatggcaac aacgttgcgc aaactattaa ctggcgaact
     3841 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     3901 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     3961 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     4021 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     4081 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     4141 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     4201 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     4261 cttaataaga tgatcttctt gagatcgttt tggtctgcgc gtaatctctt gctctgaaaa
     4321 cgaaaaaacc gccttgcagg gcggtttttc gaaggttctc tgagctacca actctttgaa
     4381 ccgaggtaac tggcttggag gagcgcagtc accaaaactt gtcctttcag tttagcctta
     4441 accggcgcat gacttcaaga ctaactcctc taaatcaatt accagtggct gctgccagtg
     4501 gtgcttttgc atgtctttcc gggttggact caagacgata gttaccggat aaggcgcagc
     4561 ggtcggactg aacggggggt tcgtgcatac agtccagctt ggagcgaact gcctacccgg
     4621 aactgagtgt caggcgtgga atgagacaaa cgcggccata acagcggaat gacaccggta
     4681 aaccgaaagg caggaacagg agagcgcacg agggagccgc cagggggaaa cgcctggtat
     4741 ctttatagtc ctgtcgggtt tcgccaccac tgatttgagc gtcagatttc gtgatgcttg
     4801 tcaggggggc ggagcctatg gaaaaacggc tttgccgcgg ccctctcact tccctgttaa
     4861 gtatcttcct ggcatcttcc aggaaatctc cgccccgttc gtaagccatt tccgctcgcc
     4921 gcagtcgaac gaccgagcgt agcgagtcag tgagcgagga agcggaatat atcctgtatc
     4981 acatattctg ctgacgcacc ggtgcagcct tttttctcct gccacatgaa gcacttcact
     5041 gacaccctca tcagtgccaa catagtaagc cagtatacac tccgctagcg ctgaggtctg
     5101 cctcgtgaag aaggtgttgc tgactcatac caggtagtcc gaggagcttt atgcttgtaa
     5161 accgttttgt gaaaaaattt ttaaaataaa aaaggggacc tctagggtcc ccaattaatt
     5221 agtaatataa tctattaaag gtcattcaaa aggtcatcca ccggatcaat tcccctgctc
     5281 gcgcaggctg ggtgccaagc tctcgggtaa catcaaggcc cgatccttgg agcccttgcc
     5341 ctcccgcacg atgatcgtgc cgtgatcgaa atccagatcc ttgacccgca gttgcaaacc
     5401 ctcactgatc cgctagtccg aggcctcgag ttactactgg aacaggtggt ggcgggcctc
     5461 ggcgcgctcg tactgctcca ccacggtgta gtcctcgttg tgggaggtga tgtcgagctt
     5521 gtagtccacg tagtggtagc cgggcagctt cacgggcttc ttggccatgt agatggactt
     5581 gaactcacac aggtagtggc cgccgccctt cagcttcagc gccatgtggt tctctccctt
     5641 cagcacgccg tcgcgggggt agttgcgctc agtggagggc tcccagccca gagtcttctt
     5701 ctgcattacg gggccgtcgg aggggaagtt cacgccgatg aacttcacgt ggtagatgag
     5761 ggtgccgtcc tgcagggagg agtcctgggt cacggtcacc acgccgccgt cctcgaagtt
     5821 catcacgcgc tcccacttga agccctcggg gaaggactgc ttgaggtagt cggggatgtc
     5881 ggcggggtgc ttgatgtacg ccttggagcc gtagaagaac tggggggaca ggatgtccca
     5941 ggcgaagggc agggggccgc ccttggtcac ttgcagcttg gcggtctggg tgccctcgta
     6001 gggcttgccc tcgcccacgc cctcgatctc gaactcgtgg ccgttcacgg agccctccat
     6061 gtgcaccttg aagcgcatga agggcttgat gacgttctca gtggaattca tatgtatatc
     6121 tccttcttaa agttaaacaa aattatttca agaggatcga tcccccgtct ctctaaaatt
     6181 gcagtcatgg taaaatcttg gtttatttaa tttaatcatc agggactccc aagcacatta
     6241 attcttctca aaatagagaa ttgatagctt gttattcaac agtataacat gtcttatacg
     6301 cccgtgtcaa ccaatattca ttgagatcca cgcgtcgacg tcatatggat ccgatatcgc
     6361 cgtggcggcc agcttttgcc attctcaccg gattcagtcg tcactcatgg tgatttctca
     6421 cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt tggacgagtc
     6481 ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg tgagttttct
     6541 ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga tatgaataaa
     6601 ttgcagtttc atttgatgct cgatgagttt ttctaatcag aattggttaa ttggttgtaa
     6661 cactggcaga gcattacgct gacttgacgg gacggcggct ttgttgaata aatcgaactt
     6721 ttgctgagtt gaaggatcag atcacgcatc ttcccgacaa cgcagaccgt tccgtggcaa
     6781 agcaaaagtt caaaatcacc aactggtcca cctacaacaa agctctcatc aaccgtggct
     6841 ccctcacttt ctggctggat gatggggcga ttcaggcctg gtatgagtca gcaacacctt
     6901 cttcacgagg cagacctcag cgccagaagg ccgccagaga ggccgagcgc ggccgtgagg
     6961 cttggacgct agggcagggc atgaaaaagc ccgtagcggg ctgctacggg cgtctgacgc
     7021 ggtggaaagg gggaggggat gttgtctaca tggctctgct gtagtgagtg ggttgcgctc
     7081 cggcagcggt cctgatcaat cgtcaccctt tctcggtcct tcaacgttcc tgacaacgag
     7141 cctccttttc gccaatccat cgacaatcac cgcgagtccc tgctcgaacg ctgcgtccgg
     7201 accggcttcg tcgaaggcgt ctatcgcggc ccgcaacagc ggcgagagcg gagcctgttc
     7261 aacggtgccg ccgcgctcgc cggcatcgct gtcgccggcc tgctcctcaa gcacggcccc
     7321 aacagtgaag tagctgattg tcatcagcgc attgacggcg tccccggccg aaaaacccgc
     7381 ctcgcagagg aagcgaagct gcgcgtcggc cgtttccatc tgcggtgcgc ccggtcgcgt
     7441 gccggcatgg atgcgcgcgc catcgcggta ggcgagcagc gcctgcctga agctgcgggc
     7501 attcccgatc agaaatgagc gccagtcgtc gtcggctctc ggcaccgaat gcgtatgatt
     7561 ctccgccagc atggcttcgg ccagtgcgtc gagcagcgcc cgcttgttcc tgaagtgcca
     7621 gtaaagcgcc ggctgctgaa cccccaaccg ttccgccagt ttgcgtgtcg tcagaccgtc
     7681 tacgccgacc tcgttcaaca ggtccagggc ggcacggatc actgtattcg gctgcaactt
     7741 tgtcatgctt gacactttat cactgataaa cataatatgt ccaccaactt atcagtgata
     7801 aagaatccgc gcgttcaatc ggaccagcgg aggctggtcc ggaggccaga cgtgaaaccc
     7861 aacatacccc tgatcgtaat tctgagcact gtcgcgctcg acgctgtcgg catcggcctg
     7921 attatgccgg tgctgccggg cctcctgcgc gatctggttc actcgaacga cgtcaccgcc
     7981 cactatggca ttctgctggc gctgtatgcg ttggtgcaat ttgcctgcgc acctgtgctg
     8041 ggcgcgctgt cggatcgttt cgggcggcgg ccaatcttgc tcgtctcgct ggccggcgcc
     8101 actgtcgact acgccatcat ggcgacagcg cctttccttt gggttctcta tatcgggcgg
     8161 atcgtggccg gcatcaccgg ggcgactggg gcggtagccg gcgcttatat tgccgatatc
     8221 actgatggcg atgagcgcgc gcggcacttc ggcttcatga gcgcctgttt cgggttcggg
     8281 atggtcgcgg gacctgtgct cggtgggctg atgggcggtt tctcccccca cgctccgttc
     8341 ttcgccgcgg cagccttgaa cggcctcaat ttcctgacgg gctgtttcct tttgccggag
     8401 tcgcacaaag gcgaacgccg gccgttacgc cgggaggctc tcaacccgct cgcttcgttc
     8461 cggtgggccc ggggcatgac cgtcgtcgcc gccctgatgg cggtcttctt catcatgcaa
     8521 cttgtcggac aggtgccggc cgcgctttgg gtcattttcg gcgaggatcg ctttcactgg
     8581 gacgcgacca cgatcggcat ttcgcttgcc gcatttggca ttctgcattc actcgcccag
     8641 gcaatgatca ccggccctgt agccgcccgg ctcggcgaaa ggcgggcact catgctcgga
     8701 atgattgccg acggcacagg ctacatcctg cttgccttcg cgacacgggg atggatggcg
     8761 ttcccgatca tggtcctgct tgcttcgggt ggcatcggaa tgccggcgct gcaagcaatg
     8821 ttgtccaggc aggtggatga ggaacgtcag gggcagctgc aaggctcact ggcggcgctc
     8881 accagcctga cctcgatcgt cggacccctc ctcttcacgg cgatctatgc ggcttctata
     8941 acaacgtgga acgggtgggc atggattgca ggcgctgccc tctacttgct ctgcctgccg
     9001 gcgctgcgtc gcgggctttg gagcggcgca gggcaacgag ccgatcgctg atcgtggaaa
     9061 cgataggggc cgctctagta ctagctagaa ctat
//
