LOCUS       sequence-246133-        7352 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector for expression of plastid 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 7352)
  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 7352)
  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: pTEI165
FEATURES             Location/Qualifiers
     source          1..7352
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             308..949
                     /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      980..1687
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     misc_feature    1736..1760
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(1916..3064)
                     /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(3400..3988)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3480..3499)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4159..4950,4951..5019))
                     /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     4782..4801
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5020..5124)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5211..5842
                     /label=oriV
                     /note="incP origin of replication"
     misc_feature    5934..5958
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        join(7056..7352,1..49)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     join(7344..7352,1..12)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
ORIGIN
        1 gcaagaccct tcctctatat aaggaagttc atttcatttg gagaggacac gctcgagtat
       61 aagagctcat ttttacaaca attaccaaca acaacaaaca acaaacaaca ttacaattac
      121 atttacaatt atcgatacca tgaccaccgc tgtcaccgcc gctgtttctt tcccctctac
      181 caaaaccacc tctctctccg cccgaagctc ctccgtcatt tcccctgaca aaatcagcta
      241 caaaaaggtt cctttgtact acaggaatgt atctgcaact gggaaaatgg gacccatcag
      301 ggccgcaatg tccaaaggag aagaactgtt taccggtgtt gtgccaattt tggttgaact
      361 cgatggtgat gtcaacggac ataagttctc agtgagaggc gaaggagaag gtgacgccac
      421 cattggaaaa ttgactctta aattcatctg tactactggt aaacttcctg taccatggcc
      481 gactctcgta acaacgcttg ggtacggagt tcagtgcttt tcgagatacc cagaccatat
      541 gaaaagacat gactttttta agtcggctat gcctgaaggt tacgtgcaag aaagaacaat
      601 ttcgttcaaa gatgatggaa aatataaaac tagagcagtt gttaaatttg aaggagatac
      661 tttggttaac cgcattgaac tgaaaggaac agattttaaa gaagatggta atattcttgg
      721 acacaaactc gaatacaatt ttaatagtca taacgtatac atcactgctg ataagcaaaa
      781 gaacggaatt aaagcgaatt tcacagtacg ccataatgta gaagatggca gtgttcaact
      841 tgccgaccat taccaacaaa acacccctat tggagacggt ccggtacttc ttcctgataa
      901 tcactacctc tcatatcaat cagtcctgag caaagatcca aatgaaaaag gaacaggtgg
      961 cggcggaagt tagcttgtcc tgctttaatg agatatgcga gaagcctatg atcgcatgat
     1021 atttgctttc aattctgttg tgcacgttgt aaaaaacctg agcatgtgta gctcagatcc
     1081 ttaccgccgg tttcggttca ttctaatgaa tatatcaccc gttactatcg tatttttatg
     1141 aataatattc tccgttcaat ttactgattg taccctacta cttatatgta caatattaaa
     1201 atgaaaacaa tatattgtgc tgaataggtt tatagcgaca tctatgatag agcgccacaa
     1261 taacaaacaa ttgcgtttta ttattacaaa tccaatttta aaaaaagcgg cagaaccggt
     1321 caaacctaaa agactgatta cataaatctt attcaaattt caaaagtgcc ccaggggcta
     1381 gtatctacga cacaccgagc ggcgaactaa taacgctcac tgaagggaac tccggttccc
     1441 cgccggcgcg catgggtgag attccttgaa gttgagtatt ggccgtccgc tctaccgaaa
     1501 gttacgggca ccattcaacc cggtccagca cggcggccgg gtaaccgact tgctgccccg
     1561 agaattatgc agcatttttt tggtgtatgt gggccccaaa tgaagtgcag gtcaaacctt
     1621 gacagtgacg acaaatcgtt gggcgggtcc agggcgaatt ttgcgacaac atgtcgaggc
     1681 tcagcaggac cgctactatt gtcttctgca cgaagtggtt taaactatca gtgtttgaca
     1741 ggatatattg gcgggtaaac ctaagagaaa agagcgttta ttagaataat cggatattta
     1801 aaagggcgtg aaaaggttta tccgttcgtc catttgtatg tgcatgccaa ccacagggtt
     1861 ccccagatca ggcgctggct gctgaacccc cagccggaac tgaccccaca aggccctagc
     1921 gtttgcaatg caccaggtca tcattgaccc aggcgtgttc caccaggccg ctgcctcgca
     1981 actcttcgca ggcttcgccg acctgctcgc gccacttctt cacgcgggtg gaatccgatc
     2041 cgcacatgag gcggaaggtt tccagcttga gcgggtacgg ctcccggtgc gagctgaaat
     2101 agtcgaacat ccgtcgggcc gtcggcgaca gcttgcggta cttctcccat atgaatttcg
     2161 tgtagtggtc gccagcaaac agcacgacga tttcctcgtc gatcaggacc tggcaacggg
     2221 acgttttctt gccacggtcc aggacgcgga agcggtgcag cagcgacacc gattccaggt
     2281 gcccaacgcg gtcggacgtg aagcccatcg ccgtcgcctg taggcgcgac aggcattcct
     2341 cggccttcgt gtaataccgg ccattgatcg accagcccag gtcctggcaa agctcgtaga
     2401 acgtgaaggt gatcggctcg ccgatagggg tgcgcttcgc gtactccaac acctgctgcc
     2461 acaccagttc gtcatcgtcg gcccgcagct cgacgccggt gtaggtgatc ttcacgtcct
     2521 tgttgacgtg gaaaatgacc ttgttttgca gcgcctcgcg cgggattttc ttgttgcgcg
     2581 tggtgaacag ggcagagcgg gccgtgtcgt ttggcatcgc tcgcatcgtg tccggccacg
     2641 gcgcaatatc gaacaaggaa agctgcattt ccttgatctg ctgcttcgtg tgtttcagca
     2701 acgcggcctg cttggcctcg ctgacctgtt ttgccaggtc ctcgccggcg gtttttcgct
     2761 tcttggtcgt catagttcct cgcgtgtcga tggtcatcga cttcgccaaa cctgccgcct
     2821 cctgttcaag acgacgcgaa cgctccacgg cggccgatgg cgcgggcagg gcagggggag
     2881 ccagttgcac gctgtcgcgc tcgatcttgg ccgtagcttg ctggaccatc gagccgacgg
     2941 actggaaggt ttcgcggggc gcacgcatga cggtgcggct tgcgatggtt tcggcatcct
     3001 cggcggaaaa ccccgcgtcg atcagttctt gcctgtatgc cttccggtca aacgtccgat
     3061 tcattcaccc tccttgcggg attgccccga ctcacgccgg ggcaatgtgc ccttattcct
     3121 gatttgaccc gcctggtgcc ttggtgtcca gataatccac cttatcggca atgaagtcgg
     3181 tcccgtagac cgtctggccg tccttctcgt acttggtatt ccgaatcttg ccctgcacga
     3241 ataccagcga ccccttgccc aaatacttgc cgtgggcctc ggcctgagag ccaaaacact
     3301 tgatgcggaa gaagtcggtg cgctcctgct tgtcgccggc atcgttgcgc cacatctagg
     3361 atctgccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     3421 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     3481 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     3541 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     3601 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     3661 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     3721 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     3781 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     3841 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     3901 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     3961 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     4021 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     4081 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     4141 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     4201 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     4261 gggcttacca tctggcccca gtgctgcaat gataccgcga gaaccacgct caccggctcc
     4321 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     4381 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     4441 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     4501 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     4561 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     4621 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     4681 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     4741 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     4801 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     4861 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     4921 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     4981 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     5041 ttgaagcatt tatcaaggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     5101 aaataaacaa ataggggttc cgcgcacgaa ttggccagcg ctgccatttt tggggtgagg
     5161 ccgttcgcgg ccgaggggcg cagcccctgg ggggatggga ggcccgcgtt agcgggccgg
     5221 gagggttcga gaaggggggg cacccccctt cggcgtgcgc ggtcacgcgc acagggcgca
     5281 gccctggtta aaaacaaggt ttataaatat tggtttaaaa gcaggttaaa agacaggtta
     5341 gcggtggccg aaaaacgggc ggaaaccctt gcaaatgctg gattttctgc ctgtggacag
     5401 cccctcaaat gtcaataggt gcgcccctca tctgtcagca ctctgcccct caagtgtcaa
     5461 ggatcgcgcc cctcatctgt cagtagtcgc gcccctcaag tgtcaatacc gcagggcact
     5521 tatccccagg cttgtccaca tcatctgtgg gaaactcgcg taaaatcagg cgttttcgcc
     5581 gatttgcgag gctggccagc tccacgtcgc cggccgaaat cgagcctgcc cctcatctgt
     5641 caacgccgcg ccgggtgagt cggcccctca agtgtcaacg tccgcccctc atctgtcagt
     5701 gagggccaag ttttccgcga ggtatccaca acgccggcgg ccgcggtgtc tcgcacacgg
     5761 cttcgacggc gtttctggcg cgtttgcagg gccatagacg gccgccagcc cagcggcgag
     5821 ggcaaccagc ccggtgagcg tcgcaaagga gatcctgatc tgactgatgg gctgcctgta
     5881 tcgagtggtg attttgtgcc gagctgccgg tcggggagct gttggctggc tggtggcagg
     5941 atatattgtg gtgtaaacaa attgacgctt agacaactta ataacacatt gcggacgttt
     6001 ttaatgtact ggggtggatg cagtgggccc cactctgtga agacaagcaa gaattcaagc
     6061 ttggaggaat tccaatccca caaaaatctg agcttaacag cacagttgct cctctcagag
     6121 cagaatcggg tattcaacac cctcatatca actactacgt tgtgtataac ggtccacatg
     6181 ccggtatata cgatgactgg ggttgtacaa aggcggcaac aaacggcgtt cccggagttg
     6241 cacacaagaa atttgccact attacagagg caagagcagc agctgacgcg tacacaacaa
     6301 gtcagcaaac agacaggttg aacttcatcc ccaaaggaga agctcaactc aagcccaaga
     6361 gctttgctaa ggccctaaca agcccaccaa agcaaaaagc ccactggctc acgctaggaa
     6421 ccaaaaggcc cagcagtgat ccagccccaa aagagatctc ctttgccccg gagattacaa
     6481 tggacgattt cctctatctt tacgatctag gaaggaagtt cgaaggtgaa ggtgacgaca
     6541 ctatgttcac cactgataat gagaaggtta gcctcttcaa tttcagaaag aatgctgacc
     6601 cacagatggt tagagaggcc tacgcagcaa gtctcatcaa gacgatctac ccgagtaaca
     6661 atctccagga gatcaaatac cttcccaaga aggttaaaga tgcagtcaaa agattcagga
     6721 ctaattgcat caagaacaca gagaaagaca tatttctcaa gatcagaagt actattccag
     6781 tatggacgat tcaaggcttg cttcataaac caaggcaagt aatagagatt ggagtctcta
     6841 aaaaggtagt tcctactgaa tctaaggcca tgcatggagt ctaagattca aatcgaggat
     6901 ctaacagaac tcgccgtcaa gactggcgaa cagttcatac agagtctttt acgactcaat
     6961 gacaagaaga aaatcttcgt caacatggtg gagcacgaca ctctggtcta ctccaaaaat
     7021 gtcaaagata cagtctcaga agatcaaagg gctattgaga cttttcaaca aaggataatt
     7081 tcgggaaacc tcctcggatt ccattgccca gctatctgtc acttcatcga aaggacagta
     7141 gaaaaggaag gtggctccta caaatgccat cattgcgata aaggaaaggc tatcattcaa
     7201 gatctctctg ccgacagtgg tcccaaagat ggacccccac ccacgaggag catcgtggaa
     7261 aaagaagagg ttccaaccac gtctacaaag caagtggatt gatgtgacat ctccactgac
     7321 gtaagggatg acgcacaatc ccactatcct tc
//
