LOCUS       sequence-246135-        7487 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector for expression of mitochondria targeted improved YFP1-10 
            (T65G, T205S, T203Y) 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: pTEI166
FEATURES             Location/Qualifiers
     source          1..7487
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..346
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     289..309
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             740..1381
                     /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
                     FICTTGKLPVPWPTLVTTLGYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     KYKTRAVVKFEGDTLVNRIELKGTDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFTVRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSVLSKDPNEK"
     terminator      1412..2119
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     misc_feature    2168..2192
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(2348..3496)
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     rep_origin      complement(3832..4420)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3912..3931)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4591..5382,5383..5451))
                     /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     5214..5233
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5452..5556)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5643..6274
                     /label=oriV
                     /note="incP origin of replication"
     misc_feature    6366..6390
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 tgagactttt caacaaagga taatttcggg aaacctcctc ggattccatt gcccagctat
       61 ctgtcacttc atcgaaagga cagtagaaaa ggaaggtggc tcctacaaat gccatcattg
      121 cgataaagga aaggctatca ttcaagatct ctctgccgac agtggtccca aagatggacc
      181 cccacccacg aggagcatcg tggaaaaaga agaggttcca accacgtcta caaagcaagt
      241 ggattgatgt gacatctcca ctgacgtaag ggatgacgca caatcccact atccttcgca
      301 agacccttcc tctatataag gaagttcatt tcatttggag aggacacgct cgagtataag
      361 agctcatttt tacaacaatt accaacaaca acaaacaaca aacaacatta caattacatt
      421 tacaattatc gataccatgc ttcgagtagc aggtagaagg ctttcttctt cagccgctag
      481 atcttcatct accttcttta caagaagctc tttcaccgtt accgatgatt cgtctccggc
      541 aagatctcct tctccgtcac tcacctcttc gtttctcgat caaatcagag gtttctcatc
      601 taattcggtt tctcccgcac atcagttggg tttagtctca gatcttccag ccacagtggc
      661 tgctattaag aatcccagtt caaaaattgt atatgatgac tccaaccatg agcgttatcc
      721 acctggagat cctagcgcaa tgtccaaagg agaagaactg tttaccggtg ttgtgccaat
      781 tttggttgaa ctcgatggtg atgtcaacgg acataagttc tcagtgagag gcgaaggaga
      841 aggtgacgcc accattggaa aattgactct taaattcatc tgtactactg gtaaacttcc
      901 tgtaccatgg ccgactctcg taacaacgct tgggtacgga gttcagtgct tttcgagata
      961 cccagaccat atgaaaagac atgacttttt taagtcggct atgcctgaag gttacgtgca
     1021 agaaagaaca atttcgttca aagatgatgg aaaatataaa actagagcag ttgttaaatt
     1081 tgaaggagat actttggtta accgcattga actgaaagga acagatttta aagaagatgg
     1141 taatattctt ggacacaaac tcgaatacaa ttttaatagt cataacgtat acatcactgc
     1201 tgataagcaa aagaacggaa ttaaagcgaa tttcacagta cgccataatg tagaagatgg
     1261 cagtgttcaa cttgccgacc attaccaaca aaacacccct attggagacg gtccggtact
     1321 tcttcctgat aatcactacc tctcatatca atcagtcctg agcaaagatc caaatgaaaa
     1381 aggaacaggt ggcggcggaa gttagcttgt cctgctttaa tgagatatgc gagaagccta
     1441 tgatcgcatg atatttgctt tcaattctgt tgtgcacgtt gtaaaaaacc tgagcatgtg
     1501 tagctcagat ccttaccgcc ggtttcggtt cattctaatg aatatatcac ccgttactat
     1561 cgtattttta tgaataatat tctccgttca atttactgat tgtaccctac tacttatatg
     1621 tacaatatta aaatgaaaac aatatattgt gctgaatagg tttatagcga catctatgat
     1681 agagcgccac aataacaaac aattgcgttt tattattaca aatccaattt taaaaaaagc
     1741 ggcagaaccg gtcaaaccta aaagactgat tacataaatc ttattcaaat ttcaaaagtg
     1801 ccccaggggc tagtatctac gacacaccga gcggcgaact aataacgctc actgaaggga
     1861 actccggttc cccgccggcg cgcatgggtg agattccttg aagttgagta ttggccgtcc
     1921 gctctaccga aagttacggg caccattcaa cccggtccag cacggcggcc gggtaaccga
     1981 cttgctgccc cgagaattat gcagcatttt tttggtgtat gtgggcccca aatgaagtgc
     2041 aggtcaaacc ttgacagtga cgacaaatcg ttgggcgggt ccagggcgaa ttttgcgaca
     2101 acatgtcgag gctcagcagg accgctacta ttgtcttctg cacgaagtgg tttaaactat
     2161 cagtgtttga caggatatat tggcgggtaa acctaagaga aaagagcgtt tattagaata
     2221 atcggatatt taaaagggcg tgaaaaggtt tatccgttcg tccatttgta tgtgcatgcc
     2281 aaccacaggg ttccccagat caggcgctgg ctgctgaacc cccagccgga actgacccca
     2341 caaggcccta gcgtttgcaa tgcaccaggt catcattgac ccaggcgtgt tccaccaggc
     2401 cgctgcctcg caactcttcg caggcttcgc cgacctgctc gcgccacttc ttcacgcggg
     2461 tggaatccga tccgcacatg aggcggaagg tttccagctt gagcgggtac ggctcccggt
     2521 gcgagctgaa atagtcgaac atccgtcggg ccgtcggcga cagcttgcgg tacttctccc
     2581 atatgaattt cgtgtagtgg tcgccagcaa acagcacgac gatttcctcg tcgatcagga
     2641 cctggcaacg ggacgttttc ttgccacggt ccaggacgcg gaagcggtgc agcagcgaca
     2701 ccgattccag gtgcccaacg cggtcggacg tgaagcccat cgccgtcgcc tgtaggcgcg
     2761 acaggcattc ctcggccttc gtgtaatacc ggccattgat cgaccagccc aggtcctggc
     2821 aaagctcgta gaacgtgaag gtgatcggct cgccgatagg ggtgcgcttc gcgtactcca
     2881 acacctgctg ccacaccagt tcgtcatcgt cggcccgcag ctcgacgccg gtgtaggtga
     2941 tcttcacgtc cttgttgacg tggaaaatga ccttgttttg cagcgcctcg cgcgggattt
     3001 tcttgttgcg cgtggtgaac agggcagagc gggccgtgtc gtttggcatc gctcgcatcg
     3061 tgtccggcca cggcgcaata tcgaacaagg aaagctgcat ttccttgatc tgctgcttcg
     3121 tgtgtttcag caacgcggcc tgcttggcct cgctgacctg ttttgccagg tcctcgccgg
     3181 cggtttttcg cttcttggtc gtcatagttc ctcgcgtgtc gatggtcatc gacttcgcca
     3241 aacctgccgc ctcctgttca agacgacgcg aacgctccac ggcggccgat ggcgcgggca
     3301 gggcaggggg agccagttgc acgctgtcgc gctcgatctt ggccgtagct tgctggacca
     3361 tcgagccgac ggactggaag gtttcgcggg gcgcacgcat gacggtgcgg cttgcgatgg
     3421 tttcggcatc ctcggcggaa aaccccgcgt cgatcagttc ttgcctgtat gccttccggt
     3481 caaacgtccg attcattcac cctccttgcg ggattgcccc gactcacgcc ggggcaatgt
     3541 gcccttattc ctgatttgac ccgcctggtg ccttggtgtc cagataatcc accttatcgg
     3601 caatgaagtc ggtcccgtag accgtctggc cgtccttctc gtacttggta ttccgaatct
     3661 tgccctgcac gaataccagc gaccccttgc ccaaatactt gccgtgggcc tcggcctgag
     3721 agccaaaaca cttgatgcgg aagaagtcgg tgcgctcctg cttgtcgccg gcatcgttgc
     3781 gccacatcta ggatctgcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     3841 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     3901 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     3961 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     4021 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     4081 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     4141 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     4201 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     4261 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     4321 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     4381 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     4441 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     4501 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     4561 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     4621 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     4681 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagaaccacg
     4741 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     4801 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     4861 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     4921 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     4981 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     5041 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     5101 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     5161 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     5221 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     5281 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     5341 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     5401 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     5461 ttttcaatat tattgaagca tttatcaagg ttattgtctc atgagcggat acatatttga
     5521 atgtatttag aaaaataaac aaataggggt tccgcgcacg aattggccag cgctgccatt
     5581 tttggggtga ggccgttcgc ggccgagggg cgcagcccct ggggggatgg gaggcccgcg
     5641 ttagcgggcc gggagggttc gagaaggggg ggcacccccc ttcggcgtgc gcggtcacgc
     5701 gcacagggcg cagccctggt taaaaacaag gtttataaat attggtttaa aagcaggtta
     5761 aaagacaggt tagcggtggc cgaaaaacgg gcggaaaccc ttgcaaatgc tggattttct
     5821 gcctgtggac agcccctcaa atgtcaatag gtgcgcccct catctgtcag cactctgccc
     5881 ctcaagtgtc aaggatcgcg cccctcatct gtcagtagtc gcgcccctca agtgtcaata
     5941 ccgcagggca cttatcccca ggcttgtcca catcatctgt gggaaactcg cgtaaaatca
     6001 ggcgttttcg ccgatttgcg aggctggcca gctccacgtc gccggccgaa atcgagcctg
     6061 cccctcatct gtcaacgccg cgccgggtga gtcggcccct caagtgtcaa cgtccgcccc
     6121 tcatctgtca gtgagggcca agttttccgc gaggtatcca caacgccggc ggccgcggtg
     6181 tctcgcacac ggcttcgacg gcgtttctgg cgcgtttgca gggccataga cggccgccag
     6241 cccagcggcg agggcaacca gcccggtgag cgtcgcaaag gagatcctga tctgactgat
     6301 gggctgcctg tatcgagtgg tgattttgtg ccgagctgcc ggtcggggag ctgttggctg
     6361 gctggtggca ggatatattg tggtgtaaac aaattgacgc ttagacaact taataacaca
     6421 ttgcggacgt ttttaatgta ctggggtgga tgcagtgggc cccactctgt gaagacaagc
     6481 aagaattcaa gcttggagga attccaatcc cacaaaaatc tgagcttaac agcacagttg
     6541 ctcctctcag agcagaatcg ggtattcaac accctcatat caactactac gttgtgtata
     6601 acggtccaca tgccggtata tacgatgact ggggttgtac aaaggcggca acaaacggcg
     6661 ttcccggagt tgcacacaag aaatttgcca ctattacaga ggcaagagca gcagctgacg
     6721 cgtacacaac aagtcagcaa acagacaggt tgaacttcat ccccaaagga gaagctcaac
     6781 tcaagcccaa gagctttgct aaggccctaa caagcccacc aaagcaaaaa gcccactggc
     6841 tcacgctagg aaccaaaagg cccagcagtg atccagcccc aaaagagatc tcctttgccc
     6901 cggagattac aatggacgat ttcctctatc tttacgatct aggaaggaag ttcgaaggtg
     6961 aaggtgacga cactatgttc accactgata atgagaaggt tagcctcttc aatttcagaa
     7021 agaatgctga cccacagatg gttagagagg cctacgcagc aagtctcatc aagacgatct
     7081 acccgagtaa caatctccag gagatcaaat accttcccaa gaaggttaaa gatgcagtca
     7141 aaagattcag gactaattgc atcaagaaca cagagaaaga catatttctc aagatcagaa
     7201 gtactattcc agtatggacg attcaaggct tgcttcataa accaaggcaa gtaatagaga
     7261 ttggagtctc taaaaaggta gttcctactg aatctaaggc catgcatgga gtctaagatt
     7321 caaatcgagg atctaacaga actcgccgtc aagactggcg aacagttcat acagagtctt
     7381 ttacgactca atgacaagaa gaaaatcttc gtcaacatgg tggagcacga cactctggtc
     7441 tactccaaaa atgtcaaaga tacagtctca gaagatcaaa gggctat
//
