LOCUS       sequence-246913-        7971 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Module 1 vector of PlaMiNGo toolkit for cloning of candidate gene at
            N-terminus of GFP11 and expression in plant cell.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7971)
  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 7971)
  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: pTEI161
FEATURES             Location/Qualifiers
     source          1..7971
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1023..1368
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     1311..1331
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             complement(1498..1803)
                     /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(1600..1619)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(2123..2806)
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGRKRR
                     VDPAY"
     primer_bind     2721..2740
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2807..2909)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             2940..2987
                     /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"
     misc_feature    3674..3698
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(3854..5002)
                     /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(5338..5926)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5418..5437)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6097..6888,6889..6957))
                     /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     6720..6739
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6958..7062)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7149..7780
                     /label=oriV
                     /note="incP origin of replication"
     misc_feature    7872..7896
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 tctgtgaaga caaactagaa ttcgagctcg gaggaattcc aatcccacaa aaatctgagc
       61 ttaacagcac agttgctcct ctcagagcag aatcgggtat tcaacaccct catatcaact
      121 actacgttgt gtataacggt ccacatgccg gtatatacga tgactggggt tgtacaaagg
      181 cggcaacaaa cggcgttccc ggagttgcac acaagaaatt tgccactatt acagaggcaa
      241 gagcagcagc tgacgcgtac acaacaagtc agcaaacaga caggttgaac ttcatcccca
      301 aaggagaagc tcaactcaag cccaagagct ttgctaaggc cctaacaagc ccaccaaagc
      361 aaaaagccca ctggctcacg ctaggaacca aaaggcccag cagtgatcca gccccaaaag
      421 agatctcctt tgccccggag attacaatgg acgatttcct ctatctttac gatctaggaa
      481 ggaagttcga aggtgaaggt gacgacacta tgttcaccac tgataatgag aaggttagcc
      541 tcttcaattt cagaaagaat gctgacccac agatggttag agaggcctac gcagcaagtc
      601 tcatcaagac gatctacccg agtaacaatc tccaggagat caaatacctt cccaagaagg
      661 ttaaagatgc agtcaaaaga ttcaggacta attgcatcaa gaacacagag aaagacatat
      721 ttctcaagat cagaagtact attccagtat ggacgattca aggcttgctt cataaaccaa
      781 ggcaagtaat agagattgga gtctctaaaa aggtagttcc tactgaatct aaggccatgc
      841 atggagtcta agattcaaat cgaggatcta acagaactcg ccgtcaagac tggcgaacag
      901 ttcatacaga gtcttttacg actcaatgac aagaagaaaa tcttcgtcaa catggtggag
      961 cacgacactc tggtctactc caaaaatgtc aaagatacag tctcagaaga tcaaagggct
     1021 attgagactt ttcaacaaag gataatttcg ggaaacctcc tcggattcca ttgcccagct
     1081 atctgtcact tcatcgaaag gacagtagaa aaggaaggtg gctcctacaa atgccatcat
     1141 tgcgataaag gaaaggctat cattcaagat ctctctgccg acagtggtcc caaagatgga
     1201 cccccaccca cgaggagcat cgtggaaaaa gaagaggttc caaccacgtc tacaaagcaa
     1261 gtggattgat gtgacatctc cactgacgta agggatgacg cacaatccca ctatccttcg
     1321 caagaccctt cctctatata aggaagttca tttcatttgg agaggacacg ctcgagtata
     1381 agagctcatt tttacaacaa ttaccaacaa caacaaacaa caaacaacat tacaattaca
     1441 tttacaatta tcgatacaat gtgagaccga ctggctgtgt ataagggagc ctgacattta
     1501 tattccccag aacatcaggt taatggcgtt tttgatgtca ttttcgcggt ggctgagatc
     1561 agccacttct tccccgataa cggaaaccgg cacactggcc atatcggtgg tcatcatgcg
     1621 ccagctttca tccccgatat gcaccaccgg gtaaagttca cgggagactt tatctgacag
     1681 cagacgtgca ctggccaggg ggatcaccat ccgtcgccca ggcgtgtcaa taatatcact
     1741 ctgtacatcc acaaacagac gataacggct ctctctttta taggtgtaaa ccttaaactg
     1801 catttcacca gcccctgttc tcgtcagcaa aagagccgtt catttcaata aaccgggcga
     1861 cctcagccat cccttcctga ttttccgctt tccagcgttc ggcacgcaga cgacgggctt
     1921 cattctgcat ggttgtgctt accagaccgg agatattgac atcatatatg ccttgagcaa
     1981 ctgatagctg tcgctgtcaa ctgtcactgt aatacgctgc ttcatagcat acctcttttt
     2041 gacatacttc gggtatacat atcagtatat attcttatac cgcaaaaatc agcgcgcaaa
     2101 tacgcatact gttatctggc ttttagtaag ccggatccac gcggcgttta cgccccccct
     2161 gccactcatc gcagtactgt tgtaattcat taagcattct gccgacatgg aagccatcac
     2221 aaacggcatg atgaacctga atcgccagcg gcatcagcac cttgtcgcct tgcgtataat
     2281 atttgcccat ggtgaaaacg ggggcgaaga agttgtccat attggccacg tttaaatcaa
     2341 aactggtgaa actcacccag ggattggctg aaacgaaaaa catattctca ataaaccctt
     2401 tagggaaata ggccaggttt tcaccgtaac acgccacatc ttgcgaatat atgtgtagaa
     2461 actgccggaa atcgtcgtgg tattcactcc agagcgatga aaacgtttca gtttgctcat
     2521 ggaaaacggt gtaacaaggg tgaacactat cccatatcac cagctcaccg tctttcattg
     2581 ccatacggaa ttccggatga gcattcatca ggcgggcaag aatgtgaata aaggccggat
     2641 aaaacttgtg cttatttttc tttacggtct ttaaaaaggc cgtaatatcc agctgaacgg
     2701 tctggttata ggtacattga gcaactgact gaaatgcctc aaaatgttct ttacgatgcc
     2761 attgggatat atcaacggtg gtatatccag tgattttttt ctccatttta gcttccttag
     2821 ctcctgaaaa tctcgataac tcaaaaaata cgcccggtag tgatcttatt tcattatggt
     2881 gaaagttgga acctcttacg tgccgatcaa ggtctcattc gatgggcggc aaattcatgc
     2941 gtgaccacat ggtccttcat gagtatgtaa atgctgctgg gattacataa gcttagagct
     3001 ttcgttcgta tcatcggttt cgacaacgtt cgtcaagttc aatgcatcag tttcattgcg
     3061 cacacaccag aatcctactg agtttgagta ttatggcatt gggaaaactg tttttcttgt
     3121 accatttgtt gtgcttgtaa tttactgtgt tttttattcg gttttcgcta tcgaactgtg
     3181 aaatggaaat ggatggagaa gagttaatga atgatatggt ccttttgttc attctcaaat
     3241 taatattatt tgttttttct cttatttgtt gtgtgttgaa tttgaaatta taagagatat
     3301 gcaaacattt tgttttgagt aaaaatgtgt caaatcgtgg cctctaatga ccgaagttaa
     3361 tatgaggagt aaaacacttg tagttgtacc attatgctta ttcactaggc aacaaatata
     3421 ttttcagacc tagaaaagct gcaaatgtta ctgaatacaa gtatgtcctc ttgtgtttta
     3481 gacatttatg aactttcctt tatgtaattt tccagaatcc ttgtcagatt ctaatcattg
     3541 ctttataatt atagttatac tcatggattt gtagttgagt atgaaaatat tttttaatgc
     3601 attttatgac ttgccaattg attgacaacg ctttacttgt cttctgcacg aagtggttta
     3661 aactatcagt gtttgacagg atatattggc gggtaaacct aagagaaaag agcgtttatt
     3721 agaataatcg gatatttaaa agggcgtgaa aaggtttatc cgttcgtcca tttgtatgtg
     3781 catgccaacc acagggttcc ccagatcagg cgctggctgc tgaaccccca gccggaactg
     3841 accccacaag gccctagcgt ttgcaatgca ccaggtcatc attgacccag gcgtgttcca
     3901 ccaggccgct gcctcgcaac tcttcgcagg cttcgccgac ctgctcgcgc cacttcttca
     3961 cgcgggtgga atccgatccg cacatgaggc ggaaggtttc cagcttgagc gggtacggct
     4021 cccggtgcga gctgaaatag tcgaacatcc gtcgggccgt cggcgacagc ttgcggtact
     4081 tctcccatat gaatttcgtg tagtggtcgc cagcaaacag cacgacgatt tcctcgtcga
     4141 tcaggacctg gcaacgggac gttttcttgc cacggtccag gacgcggaag cggtgcagca
     4201 gcgacaccga ttccaggtgc ccaacgcggt cggacgtgaa gcccatcgcc gtcgcctgta
     4261 ggcgcgacag gcattcctcg gccttcgtgt aataccggcc attgatcgac cagcccaggt
     4321 cctggcaaag ctcgtagaac gtgaaggtga tcggctcgcc gataggggtg cgcttcgcgt
     4381 actccaacac ctgctgccac accagttcgt catcgtcggc ccgcagctcg acgccggtgt
     4441 aggtgatctt cacgtccttg ttgacgtgga aaatgacctt gttttgcagc gcctcgcgcg
     4501 ggattttctt gttgcgcgtg gtgaacaggg cagagcgggc cgtgtcgttt ggcatcgctc
     4561 gcatcgtgtc cggccacggc gcaatatcga acaaggaaag ctgcatttcc ttgatctgct
     4621 gcttcgtgtg tttcagcaac gcggcctgct tggcctcgct gacctgtttt gccaggtcct
     4681 cgccggcggt ttttcgcttc ttggtcgtca tagttcctcg cgtgtcgatg gtcatcgact
     4741 tcgccaaacc tgccgcctcc tgttcaagac gacgcgaacg ctccacggcg gccgatggcg
     4801 cgggcagggc agggggagcc agttgcacgc tgtcgcgctc gatcttggcc gtagcttgct
     4861 ggaccatcga gccgacggac tggaaggttt cgcggggcgc acgcatgacg gtgcggcttg
     4921 cgatggtttc ggcatcctcg gcggaaaacc ccgcgtcgat cagttcttgc ctgtatgcct
     4981 tccggtcaaa cgtccgattc attcaccctc cttgcgggat tgccccgact cacgccgggg
     5041 caatgtgccc ttattcctga tttgacccgc ctggtgcctt ggtgtccaga taatccacct
     5101 tatcggcaat gaagtcggtc ccgtagaccg tctggccgtc cttctcgtac ttggtattcc
     5161 gaatcttgcc ctgcacgaat accagcgacc ccttgcccaa atacttgccg tgggcctcgg
     5221 cctgagagcc aaaacacttg atgcggaaga agtcggtgcg ctcctgcttg tcgccggcat
     5281 cgttgcgcca catctaggat ctgccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     5341 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     5401 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     5461 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     5521 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     5581 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     5641 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     5701 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     5761 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     5821 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     5881 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     5941 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     6001 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     6061 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     6121 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     6181 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     6241 accacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     6301 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     6361 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     6421 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     6481 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     6541 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     6601 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     6661 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     6721 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     6781 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     6841 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     6901 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     6961 cttccttttt caatattatt gaagcattta tcaaggttat tgtctcatga gcggatacat
     7021 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacgaatt ggccagcgct
     7081 gccatttttg gggtgaggcc gttcgcggcc gaggggcgca gcccctgggg ggatgggagg
     7141 cccgcgttag cgggccggga gggttcgaga agggggggca ccccccttcg gcgtgcgcgg
     7201 tcacgcgcac agggcgcagc cctggttaaa aacaaggttt ataaatattg gtttaaaagc
     7261 aggttaaaag acaggttagc ggtggccgaa aaacgggcgg aaacccttgc aaatgctgga
     7321 ttttctgcct gtggacagcc cctcaaatgt caataggtgc gcccctcatc tgtcagcact
     7381 ctgcccctca agtgtcaagg atcgcgcccc tcatctgtca gtagtcgcgc ccctcaagtg
     7441 tcaataccgc agggcactta tccccaggct tgtccacatc atctgtggga aactcgcgta
     7501 aaatcaggcg ttttcgccga tttgcgaggc tggccagctc cacgtcgccg gccgaaatcg
     7561 agcctgcccc tcatctgtca acgccgcgcc gggtgagtcg gcccctcaag tgtcaacgtc
     7621 cgcccctcat ctgtcagtga gggccaagtt ttccgcgagg tatccacaac gccggcggcc
     7681 gcggtgtctc gcacacggct tcgacggcgt ttctggcgcg tttgcagggc catagacggc
     7741 cgccagccca gcggcgaggg caaccagccc ggtgagcgtc gcaaaggaga tcctgatctg
     7801 actgatgggc tgcctgtatc gagtggtgat tttgtgccga gctgccggtc ggggagctgt
     7861 tggctggctg gtggcaggat atattgtggt gtaaacaaat tgacgcttag acaacttaat
     7921 aacacattgc ggacgttttt aatgtactgg ggtggatgca gtgggcccca c
//
