LOCUS       sequence-246127-        7487 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Module 1 vector of PlaMiNGo toolkit for expression of mitochondria 
            targeted GFP1-10 in plant cell.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7487)
  AUTHORS   Sharma M, Kretschmer C, Lampe C, Stuttmann J, Klosgen RB
  TITLE     Targeting specificity of nuclear-encoded organelle proteins with a 
            self-assembling split-fluorescent protein toolkit.
  JOURNAL   J Cell Sci. 2019 Jun 3;132(11). pii: jcs.230839. doi: 
            10.1242/jcs.230839.
  PUBMED    31085714
REFERENCE   2  (bases 1 to 7487)
  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: "J Cell 
            Sci."; date: "2019-06-3"; volume: "132(11). pii: jcs.230839. doi"; 
            pages: " 10.1242/jcs.230839"
COMMENT     Sequence Label: pTEI127
FEATURES             Location/Qualifiers
     source          1..7487
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    76..100
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      192..823
                     /label=oriV
                     /note="incP origin of replication"
     promoter        910..1014
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1015..1083,1084..1875)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1233..1252)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2046..2634
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2535..2554
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             2970..4118
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     misc_feature    4274..4298
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     terminator      complement(4347..5054)
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     CDS             complement(5085..5726)
                     /codon_start=1
                     /product="fragment of superfolder GFP consisting of the 
                     first 10 beta-strands"
                     /label=GFP(1-10)
                     /note="associates with GFP11, the 11th beta-strand, to 
                     reconstitute fluorescent GFP (Cabantous et al., 2005; 
                     Kamiyama et al., 2016)"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATIGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     KYKTRAVVKFEGDTLVNRIELKGTDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFTVRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQTVLSKDPNEK"
     promoter        complement(6120..6465)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(6157..6177)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
ORIGIN
        1 gtggggccca ctgcatccac cccagtacat taaaaacgtc cgcaatgtgt tattaagttg
       61 tctaagcgtc aatttgttta caccacaata tatcctgcca ccagccagcc aacagctccc
      121 cgaccggcag ctcggcacaa aatcaccact cgatacaggc agcccatcag tcagatcagg
      181 atctcctttg cgacgctcac cgggctggtt gccctcgccg ctgggctggc ggccgtctat
      241 ggccctgcaa acgcgccaga aacgccgtcg aagccgtgtg cgagacaccg cggccgccgg
      301 cgttgtggat acctcgcgga aaacttggcc ctcactgaca gatgaggggc ggacgttgac
      361 acttgagggg ccgactcacc cggcgcggcg ttgacagatg aggggcaggc tcgatttcgg
      421 ccggcgacgt ggagctggcc agcctcgcaa atcggcgaaa acgcctgatt ttacgcgagt
      481 ttcccacaga tgatgtggac aagcctgggg ataagtgccc tgcggtattg acacttgagg
      541 ggcgcgacta ctgacagatg aggggcgcga tccttgacac ttgaggggca gagtgctgac
      601 agatgagggg cgcacctatt gacatttgag gggctgtcca caggcagaaa atccagcatt
      661 tgcaagggtt tccgcccgtt tttcggccac cgctaacctg tcttttaacc tgcttttaaa
      721 ccaatattta taaaccttgt ttttaaccag ggctgcgccc tgtgcgcgtg accgcgcacg
      781 ccgaaggggg gtgccccccc ttctcgaacc ctcccggccc gctaacgcgg gcctcccatc
      841 cccccagggg ctgcgcccct cggccgcgaa cggcctcacc ccaaaaatgg cagcgctggc
      901 caattcgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
      961 ctcatgagac aataaccttg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     1021 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     1081 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     1141 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     1201 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     1261 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     1321 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     1381 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     1441 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     1501 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     1561 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     1621 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     1681 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg ttctcgcggt
     1741 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     1801 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     1861 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     1921 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     1981 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     2041 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     2101 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     2161 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     2221 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     2281 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     2341 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     2401 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     2461 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     2521 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     2581 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     2641 agcaacgcgg cctttttacg gttcctggca gatcctagat gtggcgcaac gatgccggcg
     2701 acaagcagga gcgcaccgac ttcttccgca tcaagtgttt tggctctcag gccgaggccc
     2761 acggcaagta tttgggcaag gggtcgctgg tattcgtgca gggcaagatt cggaatacca
     2821 agtacgagaa ggacggccag acggtctacg ggaccgactt cattgccgat aaggtggatt
     2881 atctggacac caaggcacca ggcgggtcaa atcaggaata agggcacatt gccccggcgt
     2941 gagtcggggc aatcccgcaa ggagggtgaa tgaatcggac gtttgaccgg aaggcataca
     3001 ggcaagaact gatcgacgcg gggttttccg ccgaggatgc cgaaaccatc gcaagccgca
     3061 ccgtcatgcg tgcgccccgc gaaaccttcc agtccgtcgg ctcgatggtc cagcaagcta
     3121 cggccaagat cgagcgcgac agcgtgcaac tggctccccc tgccctgccc gcgccatcgg
     3181 ccgccgtgga gcgttcgcgt cgtcttgaac aggaggcggc aggtttggcg aagtcgatga
     3241 ccatcgacac gcgaggaact atgacgacca agaagcgaaa aaccgccggc gaggacctgg
     3301 caaaacaggt cagcgaggcc aagcaggccg cgttgctgaa acacacgaag cagcagatca
     3361 aggaaatgca gctttccttg ttcgatattg cgccgtggcc ggacacgatg cgagcgatgc
     3421 caaacgacac ggcccgctct gccctgttca ccacgcgcaa caagaaaatc ccgcgcgagg
     3481 cgctgcaaaa caaggtcatt ttccacgtca acaaggacgt gaagatcacc tacaccggcg
     3541 tcgagctgcg ggccgacgat gacgaactgg tgtggcagca ggtgttggag tacgcgaagc
     3601 gcacccctat cggcgagccg atcaccttca cgttctacga gctttgccag gacctgggct
     3661 ggtcgatcaa tggccggtat tacacgaagg ccgaggaatg cctgtcgcgc ctacaggcga
     3721 cggcgatggg cttcacgtcc gaccgcgttg ggcacctgga atcggtgtcg ctgctgcacc
     3781 gcttccgcgt cctggaccgt ggcaagaaaa cgtcccgttg ccaggtcctg atcgacgagg
     3841 aaatcgtcgt gctgtttgct ggcgaccact acacgaaatt catatgggag aagtaccgca
     3901 agctgtcgcc gacggcccga cggatgttcg actatttcag ctcgcaccgg gagccgtacc
     3961 cgctcaagct ggaaaccttc cgcctcatgt gcggatcgga ttccacccgc gtgaagaagt
     4021 ggcgcgagca ggtcggcgaa gcctgcgaag agttgcgagg cagcggcctg gtggaacacg
     4081 cctgggtcaa tgatgacctg gtgcattgca aacgctaggg ccttgtgggg tcagttccgg
     4141 ctgggggttc agcagccagc gcctgatctg gggaaccctg tggttggcat gcacatacaa
     4201 atggacgaac ggataaacct tttcacgccc ttttaaatat ccgattattc taataaacgc
     4261 tcttttctct taggtttacc cgccaatata tcctgtcaaa cactgatagt ttaaaccact
     4321 tcgtgcagaa gacaatagta gcggtcctgc tgagcctcga catgttgtcg caaaattcgc
     4381 cctggacccg cccaacgatt tgtcgtcact gtcaaggttt gacctgcact tcatttgggg
     4441 cccacataca ccaaaaaaat gctgcataat tctcggggca gcaagtcggt tacccggccg
     4501 ccgtgctgga ccgggttgaa tggtgcccgt aactttcggt agagcggacg gccaatactc
     4561 aacttcaagg aatctcaccc atgcgcgccg gcggggaacc ggagttccct tcagtgagcg
     4621 ttattagttc gccgctcggt gtgtcgtaga tactagcccc tggggcactt ttgaaatttg
     4681 aataagattt atgtaatcag tcttttaggt ttgaccggtt ctgccgcttt ttttaaaatt
     4741 ggatttgtaa taataaaacg caattgtttg ttattgtggc gctctatcat agatgtcgct
     4801 ataaacctat tcagcacaat atattgtttt cattttaata ttgtacatat aagtagtagg
     4861 gtacaatcag taaattgaac ggagaatatt attcataaaa atacgatagt aacgggtgat
     4921 atattcatta gaatgaaccg aaaccggcgg taaggatctg agctacacat gctcaggttt
     4981 tttacaacgt gcacaacaga attgaaagca aatatcatgc gatcataggc ttctcgcata
     5041 tctcattaaa gcaggacaag ctaacttccg ccgccacctg ttcctttttc atttggatct
     5101 ttgctcagga ctgtttgtgt tgagaggtag tgattatcag gaagaagtac cggaccgtct
     5161 ccaatagggg tgttttgttg gtaatggtcg gcaagttgaa cactgccatc ttctacatta
     5221 tggcgtactg tgaaattcgc tttaattccg ttcttttgct tatcagcagt gatgtatacg
     5281 ttatgactat taaaattgta ttcgagtttg tgtccaagaa tattaccatc ttctttaaaa
     5341 tctgttcctt tcagttcaat gcggttaacc aaagtatctc cttcaaattt aacaactgct
     5401 ctagttttat attttccatc atctttgaac gaaattgttc tttcttgcac gtaaccttca
     5461 ggcatagccg acttaaaaaa gtcatgtctt ttcatatggt ctgggtatct cgaaaagcac
     5521 tgaactccgt acgtaagcgt tgttacgaga gtcggccatg gtacaggaag tttaccagta
     5581 gtacagatga atttaagagt caattttcca atggtggcgt caccttctcc ttcgcctctc
     5641 actgagaact tatgtccgtt gacatcacca tcgagttcaa ccaaaattgg cacaacaccg
     5701 gtaaacagtt cttctccttt ggacattgcg ctaggatctc caggtggata acgctcatgg
     5761 ttggagtcat catatacaat ttttgaactg ggattcttaa tagcagccac tgtggctgga
     5821 agatctgaga ctaaacccaa ctgatgtgcg ggagaaaccg aattagatga gaaacctctg
     5881 atttgatcga gaaacgaaga ggtgagtgac ggagaaggag atcttgccgg agacgaatca
     5941 tcggtaacgg tgaaagagct tcttgtaaag aaggtagatg aagatctagc ggctgaagaa
     6001 gaaagccttc tacctgctac tcgaagcatg gtatcgataa ttgtaaatgt aattgtaatg
     6061 ttgtttgttg tttgttgttg ttggtaattg ttgtaaaaat gagctcttat actcgagcgt
     6121 gtcctctcca aatgaaatga acttccttat atagaggaag ggtcttgcga aggatagtgg
     6181 gattgtgcgt catcccttac gtcagtggag atgtcacatc aatccacttg ctttgtagac
     6241 gtggttggaa cctcttcttt ttccacgatg ctcctcgtgg gtgggggtcc atctttggga
     6301 ccactgtcgg cagagagatc ttgaatgata gcctttcctt tatcgcaatg atggcatttg
     6361 taggagccac cttccttttc tactgtcctt tcgatgaagt gacagatagc tgggcaatgg
     6421 aatccgagga ggtttcccga aattatcctt tgttgaaaag tctcaatagc cctttgatct
     6481 tctgagactg tatctttgac atttttggag tagaccagag tgtcgtgctc caccatgttg
     6541 acgaagattt tcttcttgtc attgagtcgt aaaagactct gtatgaactg ttcgccagtc
     6601 ttgacggcga gttctgttag atcctcgatt tgaatcttag actccatgca tggccttaga
     6661 ttcagtagga actacctttt tagagactcc aatctctatt acttgccttg gtttatgaag
     6721 caagccttga atcgtccata ctggaatagt acttctgatc ttgagaaata tgtctttctc
     6781 tgtgttcttg atgcaattag tcctgaatct tttgactgca tctttaacct tcttgggaag
     6841 gtatttgatc tcctggagat tgttactcgg gtagatcgtc ttgatgagac ttgctgcgta
     6901 ggcctctcta accatctgtg ggtcagcatt ctttctgaaa ttgaagaggc taaccttctc
     6961 attatcagtg gtgaacatag tgtcgtcacc ttcaccttcg aacttccttc ctagatcgta
     7021 aagatagagg aaatcgtcca ttgtaatctc cggggcaaag gagatctctt ttggggctgg
     7081 atcactgctg ggccttttgg ttcctagcgt gagccagtgg gctttttgct ttggtgggct
     7141 tgttagggcc ttagcaaagc tcttgggctt gagttgagct tctcctttgg ggatgaagtt
     7201 caacctgtct gtttgctgac ttgttgtgta cgcgtcagct gctgctcttg cctctgtaat
     7261 agtggcaaat ttcttgtgtg caactccggg aacgccgttt gttgccgcct ttgtacaacc
     7321 ccagtcatcg tatataccgg catgtggacc gttatacaca acgtagtagt tgatatgagg
     7381 gtgttgaata cccgattctg ctctgagagg agcaactgtg ctgttaagct cagatttttg
     7441 tgggattgga attcctccaa gcttgaattc ttgcttgtct tcacaga
//
