LOCUS       sequence-246907-        8100 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Module 1 vector of PlaMiNGo toolkit for cloning of candidate gene at
            N-terminus of GFP11x3 and expression in plant cell.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8100)
  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 8100)
  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: pTEI128
FEATURES             Location/Qualifiers
     source          1..8100
                     /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"
     CDS             complement(4988..5035)
                     /codon_start=1
                     /product="11th beta-strand of superfolder GFP"
                     /label=GFP11
                     /note="associates with GFP(1-10), consisting of the first 
                     10 beta-strands, to reconstitute fluorescent GFP (Cabantous
                     et al., 2005; Kamiyama et al., 2016)"
                     /translation="RDHMVLHEYVNAAGIT"
     CDS             complement(5051..5098)
                     /codon_start=1
                     /product="11th beta-strand of superfolder GFP"
                     /label=GFP11
                     /note="associates with GFP(1-10), consisting of the first 
                     10 beta-strands, to reconstitute fluorescent GFP (Cabantous
                     et al., 2005; Kamiyama et al., 2016)"
                     /translation="RDHMVLHEYVNAAGIT"
     CDS             complement(5114..5161)
                     /codon_start=1
                     /product="11th beta-strand of superfolder GFP"
                     /label=GFP11
                     /note="associates with GFP(1-10), consisting of the first 
                     10 beta-strands, to reconstitute fluorescent GFP (Cabantous
                     et al., 2005; Kamiyama et al., 2016)"
                     /translation="RDHMVLHEYVNAAGIT"
     promoter        5192..5294
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             5295..5978
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGRKRR
                     VDPAY"
     primer_bind     complement(5361..5380)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             6298..6603
                     /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     6482..6501
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     promoter        complement(6733..7078)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(6770..6790)
                     /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 gacaagtaaa gcgttgtcaa tcaattggca agtcataaaa tgcattaaaa
     4381 aatattttca tactcaacta caaatccatg agtataacta taattataaa gcaatgatta
     4441 gaatctgaca aggattctgg aaaattacat aaaggaaagt tcataaatgt ctaaaacaca
     4501 agaggacata cttgtattca gtaacatttg cagcttttct aggtctgaaa atatatttgt
     4561 tgcctagtga ataagcataa tggtacaact acaagtgttt tactcctcat attaacttcg
     4621 gtcattagag gccacgattt gacacatttt tactcaaaac aaaatgtttg catatctctt
     4681 ataatttcaa attcaacaca caacaaataa gagaaaaaac aaataatatt aatttgagaa
     4741 tgaacaaaag gaccatatca ttcattaact cttctccatc catttccatt tcacagttcg
     4801 atagcgaaaa ccgaataaaa aacacagtaa attacaagca caacaaatgg tacaagaaaa
     4861 acagttttcc caatgccata atactcaaac tcagtaggat tctggtgtgt gcgcaatgaa
     4921 actgatgcat tgaacttgac gaacgttgtc gaaaccgatg atacgaacga aagctctaag
     4981 cttatccggt tattccggct gcattgacat attcatgtag taccatatgg tcgcgtcctc
     5041 cactaccgcc agtgatgcct gcggcgttaa cgtattcgtg gagaaccata tgatctctac
     5101 ctccagagcc acctgtaatc ccagcagcat ttacatactc atgaaggacc atgtggtcac
     5161 gcatgaattt gccgcccatc gaatgagacc ttgatcggca cgtaagaggt tccaactttc
     5221 accataatga aataagatca ctaccgggcg tattttttga gttatcgaga ttttcaggag
     5281 ctaaggaagc taaaatggag aaaaaaatca ctggatatac caccgttgat atatcccaat
     5341 ggcatcgtaa agaacatttt gaggcatttc agtcagttgc tcaatgtacc tataaccaga
     5401 ccgttcagct ggatattacg gcctttttaa agaccgtaaa gaaaaataag cacaagtttt
     5461 atccggcctt tattcacatt cttgcccgcc tgatgaatgc tcatccggaa ttccgtatgg
     5521 caatgaaaga cggtgagctg gtgatatggg atagtgttca cccttgttac accgttttcc
     5581 atgagcaaac tgaaacgttt tcatcgctct ggagtgaata ccacgacgat ttccggcagt
     5641 ttctacacat atattcgcaa gatgtggcgt gttacggtga aaacctggcc tatttcccta
     5701 aagggtttat tgagaatatg tttttcgttt cagccaatcc ctgggtgagt ttcaccagtt
     5761 ttgatttaaa cgtggccaat atggacaact tcttcgcccc cgttttcacc atgggcaaat
     5821 attatacgca aggcgacaag gtgctgatgc cgctggcgat tcaggttcat catgccgttt
     5881 gtgatggctt ccatgtcggc agaatgctta atgaattaca acagtactgc gatgagtggc
     5941 agggggggcg taaacgccgc gtggatccgg cttactaaaa gccagataac agtatgcgta
     6001 tttgcgcgct gatttttgcg gtataagaat atatactgat atgtataccc gaagtatgtc
     6061 aaaaagaggt atgctatgaa gcagcgtatt acagtgacag ttgacagcga cagctatcag
     6121 ttgctcaagg catatatgat gtcaatatct ccggtctggt aagcacaacc atgcagaatg
     6181 aagcccgtcg tctgcgtgcc gaacgctgga aagcggaaaa tcaggaaggg atggctgagg
     6241 tcgcccggtt tattgaaatg aacggctctt ttgctgacga gaacaggggc tggtgaaatg
     6301 cagtttaagg tttacaccta taaaagagag agccgttatc gtctgtttgt ggatgtacag
     6361 agtgatatta ttgacacgcc tgggcgacgg atggtgatcc ccctggccag tgcacgtctg
     6421 ctgtcagata aagtctcccg tgaactttac ccggtggtgc atatcgggga tgaaagctgg
     6481 cgcatgatga ccaccgatat ggccagtgtg ccggtttccg ttatcgggga agaagtggct
     6541 gatctcagcc accgcgaaaa tgacatcaaa aacgccatta acctgatgtt ctggggaata
     6601 taaatgtcag gctcccttat acacagccag tcggtctcac attgtatcga taattgtaaa
     6661 tgtaattgta atgttgtttg ttgtttgttg ttgttggtaa ttgttgtaaa aatgagctct
     6721 tatactcgag cgtgtcctct ccaaatgaaa tgaacttcct tatatagagg aagggtcttg
     6781 cgaaggatag tgggattgtg cgtcatccct tacgtcagtg gagatgtcac atcaatccac
     6841 ttgctttgta gacgtggttg gaacctcttc tttttccacg atgctcctcg tgggtggggg
     6901 tccatctttg ggaccactgt cggcagagag atcttgaatg atagcctttc ctttatcgca
     6961 atgatggcat ttgtaggagc caccttcctt ttctactgtc ctttcgatga agtgacagat
     7021 agctgggcaa tggaatccga ggaggtttcc cgaaattatc ctttgttgaa aagtctcaat
     7081 agccctttga tcttctgaga ctgtatcttt gacatttttg gagtagacca gagtgtcgtg
     7141 ctccaccatg ttgacgaaga ttttcttctt gtcattgagt cgtaaaagac tctgtatgaa
     7201 ctgttcgcca gtcttgacgg cgagttctgt tagatcctcg atttgaatct tagactccat
     7261 gcatggcctt agattcagta ggaactacct ttttagagac tccaatctct attacttgcc
     7321 ttggtttatg aagcaagcct tgaatcgtcc atactggaat agtacttctg atcttgagaa
     7381 atatgtcttt ctctgtgttc ttgatgcaat tagtcctgaa tcttttgact gcatctttaa
     7441 ccttcttggg aaggtatttg atctcctgga gattgttact cgggtagatc gtcttgatga
     7501 gacttgctgc gtaggcctct ctaaccatct gtgggtcagc attctttctg aaattgaaga
     7561 ggctaacctt ctcattatca gtggtgaaca tagtgtcgtc accttcacct tcgaacttcc
     7621 ttcctagatc gtaaagatag aggaaatcgt ccattgtaat ctccggggca aaggagatct
     7681 cttttggggc tggatcactg ctgggccttt tggttcctag cgtgagccag tgggcttttt
     7741 gctttggtgg gcttgttagg gccttagcaa agctcttggg cttgagttga gcttctcctt
     7801 tggggatgaa gttcaacctg tctgtttgct gacttgttgt gtacgcgtca gctgctgctc
     7861 ttgcctctgt aatagtggca aatttcttgt gtgcaactcc gggaacgccg tttgttgccg
     7921 cctttgtaca accccagtca tcgtatatac cggcatgtgg accgttatac acaacgtagt
     7981 agttgatatg agggtgttga atacccgatt ctgctctgag aggagcaact gtgctgttaa
     8041 gctcagattt ttgtgggatt ggaattcctc cgagctcgaa ttctagtttg tcttcacaga
//
