LOCUS       sequence-246910-        8349 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Module 1 vector of PlaMiNGo toolkit for cloning of candidate gene at
            N-terminus of GFP11x7 and expression in plant cell.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8349)
  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 8349)
  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: pTEI129
FEATURES             Location/Qualifiers
     source          1..8349
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1092..1437
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     1380..1400
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             complement(1567..1872)
                     /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(1669..1688)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(2192..2875)
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGRKRR
                     VDPAY"
     primer_bind     2790..2809
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2876..2978)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             3009..3434
                     /codon_start=1
                     /product="seven tandem copies of the 11th beta-strand of 
                     superfolder GFP"
                     /label=GFP11x7
                     /note="each GFP11 associates with GFP(1-10), consisting of 
                     the first 10 beta-strands, to reconstitute fluorescent GFP 
                     (Kamiyama et al., 2016)"
                     /translation="RDHMVLHEYVNAAGITGGSGGRDHMVLHEYVNAAGITGGSGGRDH
                     MVLHEYVNAAGITGGSGGRDHMVLHEYVNAAGITGGSGGRDHMVLHEYVNAAGITGGSG
                     GRDHMVLHEYVNAAGITGGSGGRDHMVLHEYVNAAGIT"
     misc_feature    4121..4145
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(4301..5449)
                     /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(5785..6373)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5865..5884)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6544..7335,7336..7404))
                     /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     7167..7186
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7405..7509)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7596..8227
                     /label=oriV
                     /note="incP origin of replication"
     misc_feature    8319..8343
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 acgcttagac aacttaataa cacattgcgg acgtttttaa tgtactgggg tggatgcagt
       61 gggccccact ctgtgaagac aaactagaat tcgagctcgg aggaattcca atcccacaaa
      121 aatctgagct taacagcaca gttgctcctc tcagagcaga atcgggtatt caacaccctc
      181 atatcaacta ctacgttgtg tataacggtc cacatgccgg tatatacgat gactggggtt
      241 gtacaaaggc ggcaacaaac ggcgttcccg gagttgcaca caagaaattt gccactatta
      301 cagaggcaag agcagcagct gacgcgtaca caacaagtca gcaaacagac aggttgaact
      361 tcatccccaa aggagaagct caactcaagc ccaagagctt tgctaaggcc ctaacaagcc
      421 caccaaagca aaaagcccac tggctcacgc taggaaccaa aaggcccagc agtgatccag
      481 ccccaaaaga gatctccttt gccccggaga ttacaatgga cgatttcctc tatctttacg
      541 atctaggaag gaagttcgaa ggtgaaggtg acgacactat gttcaccact gataatgaga
      601 aggttagcct cttcaatttc agaaagaatg ctgacccaca gatggttaga gaggcctacg
      661 cagcaagtct catcaagacg atctacccga gtaacaatct ccaggagatc aaataccttc
      721 ccaagaaggt taaagatgca gtcaaaagat tcaggactaa ttgcatcaag aacacagaga
      781 aagacatatt tctcaagatc agaagtacta ttccagtatg gacgattcaa ggcttgcttc
      841 ataaaccaag gcaagtaata gagattggag tctctaaaaa ggtagttcct actgaatcta
      901 aggccatgca tggagtctaa gattcaaatc gaggatctaa cagaactcgc cgtcaagact
      961 ggcgaacagt tcatacagag tcttttacga ctcaatgaca agaagaaaat cttcgtcaac
     1021 atggtggagc acgacactct ggtctactcc aaaaatgtca aagatacagt ctcagaagat
     1081 caaagggcta ttgagacttt tcaacaaagg ataatttcgg gaaacctcct cggattccat
     1141 tgcccagcta tctgtcactt catcgaaagg acagtagaaa aggaaggtgg ctcctacaaa
     1201 tgccatcatt gcgataaagg aaaggctatc attcaagatc tctctgccga cagtggtccc
     1261 aaagatggac ccccacccac gaggagcatc gtggaaaaag aagaggttcc aaccacgtct
     1321 acaaagcaag tggattgatg tgacatctcc actgacgtaa gggatgacgc acaatcccac
     1381 tatccttcgc aagacccttc ctctatataa ggaagttcat ttcatttgga gaggacacgc
     1441 tcgagtataa gagctcattt ttacaacaat taccaacaac aacaaacaac aaacaacatt
     1501 acaattacat ttacaattat cgatacaatg tgagaccgac tggctgtgta taagggagcc
     1561 tgacatttat attccccaga acatcaggtt aatggcgttt ttgatgtcat tttcgcggtg
     1621 gctgagatca gccacttctt ccccgataac ggaaaccggc acactggcca tatcggtggt
     1681 catcatgcgc cagctttcat ccccgatatg caccaccggg taaagttcac gggagacttt
     1741 atctgacagc agacgtgcac tggccagggg gatcaccatc cgtcgcccag gcgtgtcaat
     1801 aatatcactc tgtacatcca caaacagacg ataacggctc tctcttttat aggtgtaaac
     1861 cttaaactgc atttcaccag cccctgttct cgtcagcaaa agagccgttc atttcaataa
     1921 accgggcgac ctcagccatc ccttcctgat tttccgcttt ccagcgttcg gcacgcagac
     1981 gacgggcttc attctgcatg gttgtgctta ccagaccgga gatattgaca tcatatatgc
     2041 cttgagcaac tgatagctgt cgctgtcaac tgtcactgta atacgctgct tcatagcata
     2101 cctctttttg acatacttcg ggtatacata tcagtatata ttcttatacc gcaaaaatca
     2161 gcgcgcaaat acgcatactg ttatctggct tttagtaagc cggatccacg cggcgtttac
     2221 gccccccctg ccactcatcg cagtactgtt gtaattcatt aagcattctg ccgacatgga
     2281 agccatcaca aacggcatga tgaacctgaa tcgccagcgg catcagcacc ttgtcgcctt
     2341 gcgtataata tttgcccatg gtgaaaacgg gggcgaagaa gttgtccata ttggccacgt
     2401 ttaaatcaaa actggtgaaa ctcacccagg gattggctga aacgaaaaac atattctcaa
     2461 taaacccttt agggaaatag gccaggtttt caccgtaaca cgccacatct tgcgaatata
     2521 tgtgtagaaa ctgccggaaa tcgtcgtggt attcactcca gagcgatgaa aacgtttcag
     2581 tttgctcatg gaaaacggtg taacaagggt gaacactatc ccatatcacc agctcaccgt
     2641 ctttcattgc catacggaat tccggatgag cattcatcag gcgggcaaga atgtgaataa
     2701 aggccggata aaacttgtgc ttatttttct ttacggtctt taaaaaggcc gtaatatcca
     2761 gctgaacggt ctggttatag gtacattgag caactgactg aaatgcctca aaatgttctt
     2821 tacgatgcca ttgggatata tcaacggtgg tatatccagt gatttttttc tccattttag
     2881 cttccttagc tcctgaaaat ctcgataact caaaaaatac gcccggtagt gatcttattt
     2941 cattatggtg aaagttggaa cctcttacgt gccgatcaag gtctcattcg atgggcggca
     3001 aattcatgcg tgaccacatg gtccttcatg agtatgtaaa tgctgctggg attacaggtg
     3061 gctctggagg tagagatcat atggttctcc acgaatacgt taacgccgca ggcatcactg
     3121 gcggtagtgg aggacgcgac catatggtac tacatgaata tgtcaatgca gccggaataa
     3181 ccggagggtc cggaggccgg gatcacatgg tgctgcatga gtatgtgaac gcggcgggta
     3241 taactggtgg gtcgggcgga cgagatcata tggtgcttca cgaatacgta aacgcagctg
     3301 gcattactgg cggatcaggt ggcagggatc acatggtact ccatgagtac gtgaacgctg
     3361 ctggaatcac aggcggtagc ggcggtcggg accatatggt cctgcacgaa tatgtcaatg
     3421 ctgccggtat cacctaagct tagagctttc gttcgtatca tcggtttcga caacgttcgt
     3481 caagttcaat gcatcagttt cattgcgcac acaccagaat cctactgagt ttgagtatta
     3541 tggcattggg aaaactgttt ttcttgtacc atttgttgtg cttgtaattt actgtgtttt
     3601 ttattcggtt ttcgctatcg aactgtgaaa tggaaatgga tggagaagag ttaatgaatg
     3661 atatggtcct tttgttcatt ctcaaattaa tattatttgt tttttctctt atttgttgtg
     3721 tgttgaattt gaaattataa gagatatgca aacattttgt tttgagtaaa aatgtgtcaa
     3781 atcgtggcct ctaatgaccg aagttaatat gaggagtaaa acacttgtag ttgtaccatt
     3841 atgcttattc actaggcaac aaatatattt tcagacctag aaaagctgca aatgttactg
     3901 aatacaagta tgtcctcttg tgttttagac atttatgaac tttcctttat gtaattttcc
     3961 agaatccttg tcagattcta atcattgctt tataattata gttatactca tggatttgta
     4021 gttgagtatg aaaatatttt ttaatgcatt ttatgacttg ccaattgatt gacaacgctt
     4081 tacttgtctt ctgcacgaag tggtttaaac tatcagtgtt tgacaggata tattggcggg
     4141 taaacctaag agaaaagagc gtttattaga ataatcggat atttaaaagg gcgtgaaaag
     4201 gtttatccgt tcgtccattt gtatgtgcat gccaaccaca gggttcccca gatcaggcgc
     4261 tggctgctga acccccagcc ggaactgacc ccacaaggcc ctagcgtttg caatgcacca
     4321 ggtcatcatt gacccaggcg tgttccacca ggccgctgcc tcgcaactct tcgcaggctt
     4381 cgccgacctg ctcgcgccac ttcttcacgc gggtggaatc cgatccgcac atgaggcgga
     4441 aggtttccag cttgagcggg tacggctccc ggtgcgagct gaaatagtcg aacatccgtc
     4501 gggccgtcgg cgacagcttg cggtacttct cccatatgaa tttcgtgtag tggtcgccag
     4561 caaacagcac gacgatttcc tcgtcgatca ggacctggca acgggacgtt ttcttgccac
     4621 ggtccaggac gcggaagcgg tgcagcagcg acaccgattc caggtgccca acgcggtcgg
     4681 acgtgaagcc catcgccgtc gcctgtaggc gcgacaggca ttcctcggcc ttcgtgtaat
     4741 accggccatt gatcgaccag cccaggtcct ggcaaagctc gtagaacgtg aaggtgatcg
     4801 gctcgccgat aggggtgcgc ttcgcgtact ccaacacctg ctgccacacc agttcgtcat
     4861 cgtcggcccg cagctcgacg ccggtgtagg tgatcttcac gtccttgttg acgtggaaaa
     4921 tgaccttgtt ttgcagcgcc tcgcgcggga ttttcttgtt gcgcgtggtg aacagggcag
     4981 agcgggccgt gtcgtttggc atcgctcgca tcgtgtccgg ccacggcgca atatcgaaca
     5041 aggaaagctg catttccttg atctgctgct tcgtgtgttt cagcaacgcg gcctgcttgg
     5101 cctcgctgac ctgttttgcc aggtcctcgc cggcggtttt tcgcttcttg gtcgtcatag
     5161 ttcctcgcgt gtcgatggtc atcgacttcg ccaaacctgc cgcctcctgt tcaagacgac
     5221 gcgaacgctc cacggcggcc gatggcgcgg gcagggcagg gggagccagt tgcacgctgt
     5281 cgcgctcgat cttggccgta gcttgctgga ccatcgagcc gacggactgg aaggtttcgc
     5341 ggggcgcacg catgacggtg cggcttgcga tggtttcggc atcctcggcg gaaaaccccg
     5401 cgtcgatcag ttcttgcctg tatgccttcc ggtcaaacgt ccgattcatt caccctcctt
     5461 gcgggattgc cccgactcac gccggggcaa tgtgccctta ttcctgattt gacccgcctg
     5521 gtgccttggt gtccagataa tccaccttat cggcaatgaa gtcggtcccg tagaccgtct
     5581 ggccgtcctt ctcgtacttg gtattccgaa tcttgccctg cacgaatacc agcgacccct
     5641 tgcccaaata cttgccgtgg gcctcggcct gagagccaaa acacttgatg cggaagaagt
     5701 cggtgcgctc ctgcttgtcg ccggcatcgt tgcgccacat ctaggatctg ccaggaaccg
     5761 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     5821 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     5881 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     5941 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     6001 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     6061 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     6121 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     6181 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     6241 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     6301 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     6361 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     6421 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     6481 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     6541 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     6601 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     6661 ccccagtgct gcaatgatac cgcgagaacc acgctcaccg gctccagatt tatcagcaat
     6721 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     6781 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     6841 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     6901 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     6961 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     7021 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     7081 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     7141 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     7201 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     7261 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     7321 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     7381 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     7441 aggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     7501 ggttccgcgc acgaattggc cagcgctgcc atttttgggg tgaggccgtt cgcggccgag
     7561 gggcgcagcc cctgggggga tgggaggccc gcgttagcgg gccgggaggg ttcgagaagg
     7621 gggggcaccc cccttcggcg tgcgcggtca cgcgcacagg gcgcagccct ggttaaaaac
     7681 aaggtttata aatattggtt taaaagcagg ttaaaagaca ggttagcggt ggccgaaaaa
     7741 cgggcggaaa cccttgcaaa tgctggattt tctgcctgtg gacagcccct caaatgtcaa
     7801 taggtgcgcc cctcatctgt cagcactctg cccctcaagt gtcaaggatc gcgcccctca
     7861 tctgtcagta gtcgcgcccc tcaagtgtca ataccgcagg gcacttatcc ccaggcttgt
     7921 ccacatcatc tgtgggaaac tcgcgtaaaa tcaggcgttt tcgccgattt gcgaggctgg
     7981 ccagctccac gtcgccggcc gaaatcgagc ctgcccctca tctgtcaacg ccgcgccggg
     8041 tgagtcggcc cctcaagtgt caacgtccgc ccctcatctg tcagtgaggg ccaagttttc
     8101 cgcgaggtat ccacaacgcc ggcggccgcg gtgtctcgca cacggcttcg acggcgtttc
     8161 tggcgcgttt gcagggccat agacggccgc cagcccagcg gcgagggcaa ccagcccggt
     8221 gagcgtcgca aaggagatcc tgatctgact gatgggctgc ctgtatcgag tggtgatttt
     8281 gtgccgagct gccggtcggg gagctgttgg ctggctggtg gcaggatata ttgtggtgta
     8341 aacaaattg
//
