LOCUS       sequence-449159-        7066 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For Agrobacterium-mediated transformation. Plastid(outer envelope 
            membrane)-localized mCherry-UnaG. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7066)
  AUTHORS   Ishikawa K, Kodama Y
  TITLE     Bilirubin Distribution in Plants at the Subcellular and Tissue 
            Levels.
  JOURNAL   Plant Cell Physiol. 2024 Feb 27:pcae017. doi: 10.1093/pcp/pcae017.
  PUBMED    38466577
REFERENCE   2  (bases 1 to 7066)
  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: "Plant Cell 
            Physiol. 2024 Feb 27:pcae017. doi: 10.1093/pcp/pcae017."
COMMENT     Sequence Label: pGWB602-oeUnaG
FEATURES             Location/Qualifiers
     source          1..7066
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      420..672
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     protein_bind    708..729
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(744..749,750..767,768..774)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    782..798
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     787..809
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     806..822
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     806..822
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     terminator      complement(855..1107)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     protein_bind    1139..1263
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1406..1425)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             1865..2938
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     rep_origin      3004..3198
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(3434..3456)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3542..3682
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3597..3616
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3697..3714)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3868..4456)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3948..3967)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4702..5493)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     misc_feature    6014..6038
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        6544..6723
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     CDS             join(6775..7066,1..260)
                     /codon_start=1
                     /gene="Streptomyces hygroscopicus bar"
                     /product="phosphinothricin acetyltransferase"
                     /label=BlpR
                     /note="confers resistance to bialophos or phosphinothricin"
                     /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEI"
ORIGIN
        1 agcggacggg actgggctcc acgctctaca cccacctgct gaagtccctg gaggcacagg
       61 gcttcaagag cgtggtcgct gtcatcgggc tgcccaacga cccgagcgtg cgcatgcacg
      121 aggcgctcgg atatgccccc cgcggcatgc tgcgggcggc cggcttcaag cacgggaact
      181 ggcatgacgt gggtttctgg cagctggact tcagcctgcc ggtaccgccc cgtccggtcc
      241 tgcccgtcac cgagatctga acggagtgcg cgtggcatcg cccgagttgg agctggtacg
      301 ggaactcatc gaactcaact ggcatacccg caatggtgag gtggaaccgc ggcggatcgc
      361 gtacgaccgt gcccaggagg gggtaccgag ctggggacgt ccccagctcg aatttccccg
      421 atcgttcaaa catttggcaa taaagtttct taagattgaa tcctgttgcc ggtcttgcga
      481 tgattatcat ataatttctg ttgaattacg ttaagcatgt aataattaac atgtaatgca
      541 tgacgttatt tatgagatgg gtttttatga ttagagtccc gcaattatac atttaatacg
      601 cgatagaaaa caaaatatag cgcgcaaact aggataaatt atcgcgcgcg gtgtcatcta
      661 tgttactaga tcgggaatta atagaaatag gcgcgcctaa ttcagcctaa tgtgagttag
      721 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
      781 attgtgagcg gataacaatt tcacacagga aacagctatg acatgattac gaattggttc
      841 cggaaccaat tcccgatcta gtaacataga tgacaccgcg cgcgataatt tatcctagtt
      901 tgcgcgctat attttgtttt ctatcgcgta ttaaatgtat aattgcggga ctctaatcat
      961 aaaaacccat ctcataaata acgtcatgca ttacatgtta attattacat gcttaacgta
     1021 attcaacaga aattatatga taatcatcgc aagaccggca acaggattca atcttaagaa
     1081 actttattgc caaatgtttg aacgatcggg gaaattcgag ctctaagcgc tgttatcaac
     1141 cactttgtac aagaaagctg aacgagaaac gtaaaatgat ataaatatca atatattaaa
     1201 ttagattttg cataaaaaac agactacata atactgtaaa acacaacata tccagtcact
     1261 atggtcgacc tgcagactgg ctgtgtataa gggagcctga catttatatt ccccagaaca
     1321 tcaggttaat ggcgtttttg atgtcatttt cgcggtggct gagatcagcc acttcttccc
     1381 cgataacgga gaccggcaca ctggccatat cggtggtcat catgcgccag ctttcatccc
     1441 cgatatgcac caccgggtaa agttcacggg agactttatc tgacagcaga cgtgcactgg
     1501 ccagggggat caccatccgt cgcccgggcg tgtcaataat atcactctgt acatccacaa
     1561 acagacgatg ttgccggcgg aggatcacac caagctgaag atgtacgcgg tacgccaagg
     1621 caagaccatt accgagctgc tatctgaata catcgcgcag ctaccagagt aaatgagcaa
     1681 atgaataaat gagtagatga attttagcgg ctaaaggagg cggcatggaa aatcaagaac
     1741 aaccaggcac cgacgccgtg gaatgcccca tgtgtggagg aacgggcggt tggccaggcg
     1801 taagcggctg ggttgtctgc cggccctgca atggcactgg aacccccaag cccgaggaat
     1861 cggcgtgagc ggtcgcaaac catccggccc ggtacaaatc ggcgcggcgc tgggtgatga
     1921 cctggtggag aagttgaagg ccgcgcaggc cgcccagcgg caacgcatcg aggcagaagc
     1981 acgccccggt gaatcgtggc aagcggccgc tgatcgaatc cgcaaagaat cccggcaacc
     2041 gccggcagcc ggtgcgccgt cgattaggaa gccgcccaag ggcgacgagc aaccagattt
     2101 tttcgttccg atgctctatg acgtgggcac ccgcgatagt cgcagcatca tggacgtggc
     2161 cgttttccgt ctgtcgaagc gtgaccgacg agctggcgag gtgatccgct acgagcttcc
     2221 agacgggcac gtagaggttt ccgcagggcc ggccggcatg gccagtgtgt gggattacga
     2281 cctggtactg atggcggttt cccatctaac cgaatccatg aaccgatacc gggaagggaa
     2341 gggagacaag cccggccgcg tgttccgtcc acacgttgcg gacgtactca agttctgccg
     2401 gcgagccgat ggcggaaagc agaaagacga cctggtagaa acctgcattc ggttaaacac
     2461 cacgcacgtt gccatgcagc gtacgaagaa ggccaagaac ggccgcctgg tgacggtatc
     2521 cgagggtgaa gccttgatta gccgctacaa gatcgtaaag agcgaaaccg ggcggccgga
     2581 gtacatcgag atcgagctag ctgattggat gtaccgcgag atcacagaag gcaagaaccc
     2641 ggacgtgctg acggttcacc ccgattactt tttgatcgat cccggcatcg gccgttttct
     2701 ctaccgcctg gcacgccgcg ccgcaggcaa ggcagaagcc agatggttgt tcaagacgat
     2761 ctacgaacgc agtggcagcg ccggagagtt caagaagttc tgtttcaccg tgcgcaagct
     2821 gatcgggtca aatgacctgc cggagtacga tttgaaggag gaggcggggc aggctggccc
     2881 gatcctagtc atgcgctacc gcaacctgat cgagggcgaa gcatccgccg gttcctaatg
     2941 tacggagcag atgctagggc aaattgccct agcaggggaa aaaggtcgaa aaggtctctt
     3001 tcctgtggat agcacgtaca ttgggaaccc aaagccgtac attgggaacc ggaacccgta
     3061 cattgggaac ccaaagccgt acattgggaa ccggtcacac atgtaagtga ctgatataaa
     3121 agagaaaaaa ggcgattttt ccgcctaaaa ctctttaaaa cttattaaaa ctcttaaaac
     3181 ccgcctggcc tgtgcataac tgtctggcca gcgcacagcc gaagagctgc aaaaagcgcc
     3241 tacccttcgg tcgctgcgct ccctacgccc cgccgcttcg cgtcggccta tcgcggccgc
     3301 tggccgctca aaaatggctg gcctacggcc aggcaatcta ccagggcgcg gacaagccgc
     3361 gccgtcgcca ctcgaccgcc ggcgcccaca tcaaggcacc ctgcctcgcg cgtttcggtg
     3421 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     3481 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     3541 gcgcagccat gacccagtca cgtagcgata gcggagtgta tactggctta actatgcggc
     3601 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt
     3661 aaggagaaaa taccgcatca ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     3721 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     3781 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     3841 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     3901 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     3961 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     4021 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     4081 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     4141 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     4201 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     4261 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     4321 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     4381 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     4441 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     4501 cgaaaactca cgttaaggga ttttggtcat gcatgatata tctcccaatt tgtgtagggc
     4561 ttattatgca cgcttaaaaa taataaaagc agacttgacc tgatagtttg gctgtgagca
     4621 attatgtgct tagtgcatct aacgcttgag ttaagccgcg ccgcgaagcg gcgtcggctt
     4681 gaacgaattt ctagctagac attatttgcc gactaccttg gtgatctcgc ctttcacgta
     4741 gtggacaaat tcttccaact gatctgcgcg cgaggccaag cgatcttctt cttgtccaag
     4801 ataagcctgt ctagcttcaa gtatgacggg ctgatactgg gccggcaggc gctccattgc
     4861 ccagtcggca gcgacatcct tcggcgcgat tttgccggtt actgcgctgt accaaatgcg
     4921 ggacaacgta agcactacat ttcgctcatc gccagcccag tcgggcggcg agttccatag
     4981 cgttaaggtt tcatttagcg cctcaaatag atcctgttca ggaaccggat caaagagttc
     5041 ctccgccgct ggacctacca aggcaacgct atgttctctt gcttttgtca gcaagatagc
     5101 cagatcaatg tcgatcgtgg ctggctcgaa gatacctgca agaatgtcat tgcgctgcca
     5161 ttctccaaat tgcagttcgc gcttagctgg ataacgccac ggaatgatgt cgtcgtgcac
     5221 aacaatggtg acttctacag cgcggagaat ctcgctctct ccaggggaag ccgaagtttc
     5281 caaaaggtcg ttgatcaaag ctcgccgcgt tgtttcatca agccttacgg tcaccgtaac
     5341 cagcaaatca atatcactgt gtggcttcag gccgccatcc actgcggagc cgtacaaatg
     5401 tacggccagc aacgtcggtt cgagatggcg ctcgatgacg ccaactacct ctgatagttg
     5461 agtcgatact tcggcgatca ccgcttcccc catgatgttt aactttgttt tagggcgact
     5521 gccctgctgc gtaacatcgt tgctgctcca taacatcaaa catcgaccca cggcgtaacg
     5581 cgcttgctgc ttggatgccc gaggcataga ctgtacccca aaaaaacagt cataacaagc
     5641 catgaaaacc gccactgcgc cgttaccacc gctgcgttcg gtcaaggttc tggaccagtt
     5701 gcgtgagcgc atacgctact tgcattacag cttacgaacc gaacaggctt atgtccactg
     5761 ggttcgtgcc cgaattgatc acaggcagca acgctctgtc atcgttacaa tcaacatgct
     5821 accctccgcg agatcatccg tgtttcaaac ccggcagctt agttgccgtt cttccgaata
     5881 gcatcggtaa catgagcaaa gtctgccgcc ttacaacggc tctcccgctg acgccgtccc
     5941 ggactgatgg gctgcctgta tcgagtggtg attttgtgcc gagctgccgg tcggggagct
     6001 gttggctggc tggtggcagg atatattgtg gtgtaaacaa attgacgctt agacaactta
     6061 ataacacatt gcggacgttt ttaatgtact gaattaacgc cgaattaggc tgaatttatt
     6121 tctattaatc tcgacggatc agcttcacgc tgccgcaagc actcagggcg caagggctgc
     6181 taaaggaagc ggaacacgta gaaagccagt ccgcagaaac ggtgctgacc ccggatgaat
     6241 gtcagctact gggctatctg gacaagggaa aacgcaagcg caaagagaaa gcaggtagct
     6301 tgcagtgggc ttacatggcg atagctagac tgggcggttt tatggacagc aagcgaaccg
     6361 gaattgccag ctggggcgcc ctctggtaag gttgggaagc cctgcaaagt aaactggatg
     6421 gctttcttgc cgccaaggat ctgatggcgc aggggatcaa gatcatgagc ggagaattaa
     6481 gggagtcacg ttatgacccc cgccgatgac gcgggacaag ccgttttacg tttggaactg
     6541 acagaaccgc aacgttgaag gagccactca gccgcgggtt tctggagttt aatgagctaa
     6601 gcacatacgt cagaaaccat tattgcgcgt tcaaaagtcg cctaaggtca ctatcagcta
     6661 gcaaatattt cttgtcaaaa atgctccact gacgttccat aaattcccct cggtatccaa
     6721 ttagagtctc atattcactc tcaatccaga tctgatcccg gggggcaata agatatgagc
     6781 ccagaacgac gcccggccga catccgccgt gccaccgagg cggacatgcc ggcggtctgc
     6841 accatcgtca accactacat cgagacaagc acggtcaact tccgtaccga gccgcaggaa
     6901 ccgcaggagt ggacggacga cctcgtccgt ctgcgggagc gctatccctg gctcgtcgcc
     6961 gaggtggacg gcgaggtcgc cggcatcgcc tacgcgggcc cctggaaggc acgcaacgcc
     7021 tacgactgga cggccgagtc gaccgtgtac gtctcccccc gccacc
//
