LOCUS       sequence-246860-        8643 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Module 1 vector of PlaMiNGo toolkit for cloning of candidate gene at
            N-terminus of GFP1-10 and expression in plant cell.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8643)
  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 8643)
  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: pTEI177
FEATURES             Location/Qualifiers
     source          1..8643
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1069..1414
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     1357..1377
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             complement(1543..1848)
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     complement(1645..1664)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(2168..2851)
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGRKRR
                     VDPAY"
     primer_bind     2766..2785
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2852..2954)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             2964..3605
                     /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"
     terminator      3636..4343
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     misc_feature    4392..4416
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(4572..5720)
                     /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(6056..6644)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6136..6155)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6815..7606,7607..7675))
                     /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     7438..7457
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7676..7780)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7867..8498
                     /label=oriV
                     /note="incP origin of replication"
     misc_feature    8590..8614
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 attgcggacg tttttaatgt actggggtgg atgcagtggg ccccactctg tgaagacaag
       61 caagaattca agcttggagg aattccaatc ccacaaaaat ctgagcttaa cagcacagtt
      121 gctcctctca gagcagaatc gggtattcaa caccctcata tcaactacta cgttgtgtat
      181 aacggtccac atgccggtat atacgatgac tggggttgta caaaggcggc aacaaacggc
      241 gttcccggag ttgcacacaa gaaatttgcc actattacag aggcaagagc agcagctgac
      301 gcgtacacaa caagtcagca aacagacagg ttgaacttca tccccaaagg agaagctcaa
      361 ctcaagccca agagctttgc taaggcccta acaagcccac caaagcaaaa agcccactgg
      421 ctcacgctag gaaccaaaag gcccagcagt gatccagccc caaaagagat ctcctttgcc
      481 ccggagatta caatggacga tttcctctat ctttacgatc taggaaggaa gttcgaaggt
      541 gaaggtgacg acactatgtt caccactgat aatgagaagg ttagcctctt caatttcaga
      601 aagaatgctg acccacagat ggttagagag gcctacgcag caagtctcat caagacgatc
      661 tacccgagta acaatctcca ggagatcaaa taccttccca agaaggttaa agatgcagtc
      721 aaaagattca ggactaattg catcaagaac acagagaaag acatatttct caagatcaga
      781 agtactattc cagtatggac gattcaaggc ttgcttcata aaccaaggca agtaatagag
      841 attggagtct ctaaaaaggt agttcctact gaatctaagg ccatgcatgg agtctaagat
      901 tcaaatcgag gatctaacag aactcgccgt caagactggc gaacagttca tacagagtct
      961 tttacgactc aatgacaaga agaaaatctt cgtcaacatg gtggagcacg acactctggt
     1021 ctactccaaa aatgtcaaag atacagtctc agaagatcaa agggctattg agacttttca
     1081 acaaaggata atttcgggaa acctcctcgg attccattgc ccagctatct gtcacttcat
     1141 cgaaaggaca gtagaaaagg aaggtggctc ctacaaatgc catcattgcg ataaaggaaa
     1201 ggctatcatt caagatctct ctgccgacag tggtcccaaa gatggacccc cacccacgag
     1261 gagcatcgtg gaaaaagaag aggttccaac cacgtctaca aagcaagtgg attgatgtga
     1321 catctccact gacgtaaggg atgacgcaca atcccactat ccttcgcaag acccttcctc
     1381 tatataagga agttcatttc atttggagag gacacgctcg agtataagag ctcattttta
     1441 caacaattac caacaacaac aaacaacaaa caacattaca attacattta caattatcga
     1501 taccattgag accgactggc tgtgtataag ggagcctgac atttatattc cccagaacat
     1561 caggttaatg gcgtttttga tgtcattttc gcggtggctg agatcagcca cttcttcccc
     1621 gataacggaa accggcacac tggccatatc ggtggtcatc atgcgccagc tttcatcccc
     1681 gatatgcacc accgggtaaa gttcacggga gactttatct gacagcagac gtgcactggc
     1741 cagggggatc accatccgtc gcccaggcgt gtcaataata tcactctgta catccacaaa
     1801 cagacgataa cggctctctc ttttataggt gtaaacctta aactgcattt caccagcccc
     1861 tgttctcgtc agcaaaagag ccgttcattt caataaaccg ggcgacctca gccatccctt
     1921 cctgattttc cgctttccag cgttcggcac gcagacgacg ggcttcattc tgcatggttg
     1981 tgcttaccag accggagata ttgacatcat atatgccttg agcaactgat agctgtcgct
     2041 gtcaactgtc actgtaatac gctgcttcat agcatacctc tttttgacat acttcgggta
     2101 tacatatcag tatatattct tataccgcaa aaatcagcgc gcaaatacgc atactgttat
     2161 ctggctttta gtaagccgga tccacgcggc gtttacgccc cccctgccac tcatcgcagt
     2221 actgttgtaa ttcattaagc attctgccga catggaagcc atcacaaacg gcatgatgaa
     2281 cctgaatcgc cagcggcatc agcaccttgt cgccttgcgt ataatatttg cccatggtga
     2341 aaacgggggc gaagaagttg tccatattgg ccacgtttaa atcaaaactg gtgaaactca
     2401 cccagggatt ggctgaaacg aaaaacatat tctcaataaa ccctttaggg aaataggcca
     2461 ggttttcacc gtaacacgcc acatcttgcg aatatatgtg tagaaactgc cggaaatcgt
     2521 cgtggtattc actccagagc gatgaaaacg tttcagtttg ctcatggaaa acggtgtaac
     2581 aagggtgaac actatcccat atcaccagct caccgtcttt cattgccata cggaattccg
     2641 gatgagcatt catcaggcgg gcaagaatgt gaataaaggc cggataaaac ttgtgcttat
     2701 ttttctttac ggtctttaaa aaggccgtaa tatccagctg aacggtctgg ttataggtac
     2761 attgagcaac tgactgaaat gcctcaaaat gttctttacg atgccattgg gatatatcaa
     2821 cggtggtata tccagtgatt tttttctcca ttttagcttc cttagctcct gaaaatctcg
     2881 ataactcaaa aaatacgccc ggtagtgatc ttatttcatt atggtgaaag ttggaacctc
     2941 ttacgtgccg atcaaggtct caaatgtcca aaggagaaga actgtttacc ggtgttgtgc
     3001 caattttggt tgaactcgat ggtgatgtca acggacataa gttctcagtg agaggcgaag
     3061 gagaaggtga cgccaccatt ggaaaattga ctcttaaatt catctgtact actggtaaac
     3121 ttcctgtacc atggccgact ctcgtaacaa cgcttacgta cggagttcag tgcttttcga
     3181 gatacccaga ccatatgaaa agacatgact tttttaagtc ggctatgcct gaaggttacg
     3241 tgcaagaaag aacaatttcg ttcaaagatg atggaaaata taaaactaga gcagttgtta
     3301 aatttgaagg agatactttg gttaaccgca ttgaactgaa aggaacagat tttaaagaag
     3361 atggtaatat tcttggacac aaactcgaat acaattttaa tagtcataac gtatacatca
     3421 ctgctgataa gcaaaagaac ggaattaaag cgaatttcac agtacgccat aatgtagaag
     3481 atggcagtgt tcaacttgcc gaccattacc aacaaaacac ccctattgga gacggtccgg
     3541 tacttcttcc tgataatcac tacctctcaa cacaaacagt cctgagcaaa gatccaaatg
     3601 aaaaaggaac aggtggcggc ggaagttagc ttgtcctgct ttaatgagat atgcgagaag
     3661 cctatgatcg catgatattt gctttcaatt ctgttgtgca cgttgtaaaa aacctgagca
     3721 tgtgtagctc agatccttac cgccggtttc ggttcattct aatgaatata tcacccgtta
     3781 ctatcgtatt tttatgaata atattctccg ttcaatttac tgattgtacc ctactactta
     3841 tatgtacaat attaaaatga aaacaatata ttgtgctgaa taggtttata gcgacatcta
     3901 tgatagagcg ccacaataac aaacaattgc gttttattat tacaaatcca attttaaaaa
     3961 aagcggcaga accggtcaaa cctaaaagac tgattacata aatcttattc aaatttcaaa
     4021 agtgccccag gggctagtat ctacgacaca ccgagcggcg aactaataac gctcactgaa
     4081 gggaactccg gttccccgcc ggcgcgcatg ggtgagattc cttgaagttg agtattggcc
     4141 gtccgctcta ccgaaagtta cgggcaccat tcaacccggt ccagcacggc ggccgggtaa
     4201 ccgacttgct gccccgagaa ttatgcagca tttttttggt gtatgtgggc cccaaatgaa
     4261 gtgcaggtca aaccttgaca gtgacgacaa atcgttgggc gggtccaggg cgaattttgc
     4321 gacaacatgt cgaggctcag caggaccgct actattgtct tctgcacgaa gtggtttaaa
     4381 ctatcagtgt ttgacaggat atattggcgg gtaaacctaa gagaaaagag cgtttattag
     4441 aataatcgga tatttaaaag ggcgtgaaaa ggtttatccg ttcgtccatt tgtatgtgca
     4501 tgccaaccac agggttcccc agatcaggcg ctggctgctg aacccccagc cggaactgac
     4561 cccacaaggc cctagcgttt gcaatgcacc aggtcatcat tgacccaggc gtgttccacc
     4621 aggccgctgc ctcgcaactc ttcgcaggct tcgccgacct gctcgcgcca cttcttcacg
     4681 cgggtggaat ccgatccgca catgaggcgg aaggtttcca gcttgagcgg gtacggctcc
     4741 cggtgcgagc tgaaatagtc gaacatccgt cgggccgtcg gcgacagctt gcggtacttc
     4801 tcccatatga atttcgtgta gtggtcgcca gcaaacagca cgacgatttc ctcgtcgatc
     4861 aggacctggc aacgggacgt tttcttgcca cggtccagga cgcggaagcg gtgcagcagc
     4921 gacaccgatt ccaggtgccc aacgcggtcg gacgtgaagc ccatcgccgt cgcctgtagg
     4981 cgcgacaggc attcctcggc cttcgtgtaa taccggccat tgatcgacca gcccaggtcc
     5041 tggcaaagct cgtagaacgt gaaggtgatc ggctcgccga taggggtgcg cttcgcgtac
     5101 tccaacacct gctgccacac cagttcgtca tcgtcggccc gcagctcgac gccggtgtag
     5161 gtgatcttca cgtccttgtt gacgtggaaa atgaccttgt tttgcagcgc ctcgcgcggg
     5221 attttcttgt tgcgcgtggt gaacagggca gagcgggccg tgtcgtttgg catcgctcgc
     5281 atcgtgtccg gccacggcgc aatatcgaac aaggaaagct gcatttcctt gatctgctgc
     5341 ttcgtgtgtt tcagcaacgc ggcctgcttg gcctcgctga cctgttttgc caggtcctcg
     5401 ccggcggttt ttcgcttctt ggtcgtcata gttcctcgcg tgtcgatggt catcgacttc
     5461 gccaaacctg ccgcctcctg ttcaagacga cgcgaacgct ccacggcggc cgatggcgcg
     5521 ggcagggcag ggggagccag ttgcacgctg tcgcgctcga tcttggccgt agcttgctgg
     5581 accatcgagc cgacggactg gaaggtttcg cggggcgcac gcatgacggt gcggcttgcg
     5641 atggtttcgg catcctcggc ggaaaacccc gcgtcgatca gttcttgcct gtatgccttc
     5701 cggtcaaacg tccgattcat tcaccctcct tgcgggattg ccccgactca cgccggggca
     5761 atgtgccctt attcctgatt tgacccgcct ggtgccttgg tgtccagata atccacctta
     5821 tcggcaatga agtcggtccc gtagaccgtc tggccgtcct tctcgtactt ggtattccga
     5881 atcttgccct gcacgaatac cagcgacccc ttgcccaaat acttgccgtg ggcctcggcc
     5941 tgagagccaa aacacttgat gcggaagaag tcggtgcgct cctgcttgtc gccggcatcg
     6001 ttgcgccaca tctaggatct gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     6061 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     6121 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     6181 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     6241 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     6301 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     6361 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     6421 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     6481 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     6541 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     6601 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     6661 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     6721 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     6781 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     6841 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     6901 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagaac
     6961 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     7021 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     7081 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     7141 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     7201 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     7261 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     7321 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     7381 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     7441 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     7501 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     7561 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     7621 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     7681 tcctttttca atattattga agcatttatc aaggttattg tctcatgagc ggatacatat
     7741 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacgaattgg ccagcgctgc
     7801 catttttggg gtgaggccgt tcgcggccga ggggcgcagc ccctgggggg atgggaggcc
     7861 cgcgttagcg ggccgggagg gttcgagaag ggggggcacc ccccttcggc gtgcgcggtc
     7921 acgcgcacag ggcgcagccc tggttaaaaa caaggtttat aaatattggt ttaaaagcag
     7981 gttaaaagac aggttagcgg tggccgaaaa acgggcggaa acccttgcaa atgctggatt
     8041 ttctgcctgt ggacagcccc tcaaatgtca ataggtgcgc ccctcatctg tcagcactct
     8101 gcccctcaag tgtcaaggat cgcgcccctc atctgtcagt agtcgcgccc ctcaagtgtc
     8161 aataccgcag ggcacttatc cccaggcttg tccacatcat ctgtgggaaa ctcgcgtaaa
     8221 atcaggcgtt ttcgccgatt tgcgaggctg gccagctcca cgtcgccggc cgaaatcgag
     8281 cctgcccctc atctgtcaac gccgcgccgg gtgagtcggc ccctcaagtg tcaacgtccg
     8341 cccctcatct gtcagtgagg gccaagtttt ccgcgaggta tccacaacgc cggcggccgc
     8401 ggtgtctcgc acacggcttc gacggcgttt ctggcgcgtt tgcagggcca tagacggccg
     8461 ccagcccagc ggcgagggca accagcccgg tgagcgtcgc aaaggagatc ctgatctgac
     8521 tgatgggctg cctgtatcga gtggtgattt tgtgccgagc tgccggtcgg ggagctgttg
     8581 gctggctggt ggcaggatat attgtggtgt aaacaaattg acgcttagac aacttaataa
     8641 cac
//
