LOCUS       sequence-415310-        6868 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GreenBraid joker destination vector pGBY1j - Eco31I - dummy - 
            Eco31I.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6868)
  AUTHORS   Piepers M, Erbstein K, Reyes-Hernandez J, Song C, Tessi T, 
            Petrasiunaite V, Faerber N, Distel K, Maizel A
  TITLE     GreenGate 2.0: Backwards compatible addons for assembly of complex 
            transcriptional units and their stacking with GreenGate.
  JOURNAL   PLoS One. 2023 Sep 8;18(9):e0290097. doi: 
            10.1371/journal.pone.0290097. eCollection 2023.
  PUBMED    37682951
REFERENCE   2  (bases 1 to 6868)
  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: "PLoS One.";
            date: "2023-09-8"; volume: "18(9):e0290097. doi"; pages: " 
            10.1371/journal.pone.0290097. eCollection 2023"
COMMENT     Sequence Label: pMP074
FEATURES             Location/Qualifiers
     source          1..6868
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    204..228
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             1529..2158
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             2587..3660
                     /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      3726..3920
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(4156..4178)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4264..4404
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4319..4338
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4419..4436)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4590..5178)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4670..4689)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5266..6057)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     misc_feature    6843..6867
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 taacggatca ccatggggtc tcaacctctc aatggtaact ttactcttta tttaaccata
       61 catttttttt tatttttttc actttgttct tcatccacta ttgttctttg ttcatcttga
      121 acaaaagctc cctccttctt tgttcttcat ccaccattgt tcttcatcaa tcatttcgct
      181 gtcatgagac cctcgagaga tcctgacagg atatattggc gggtaaacct aagagaaaag
      241 agcgtttatt agaataatcg gatatttaaa agggcgtgaa aaggtttatc cgttcgtcca
      301 tttgtatgtg catgccaacc acagggttcc cctcgggatc aaagtacttt gatccaaccc
      361 ctccgctgct atagtgcagt cggcttctga cgttcagtgc agccgtcatc tgaaaacgac
      421 atgtcgcaca agtcctaagt tacgcgacag gctgccgccc tgcccttttc ctggcgtttt
      481 cttgtcgcgt gttttagtcg cataaagtag aatacttgcg actagaaccg gagacattac
      541 gccatgaaca agagcgccgc cgctggcctg ctgggctatg cccgcgtcag caccgacgac
      601 caggacttga ccaaccaacg ggccgaactg cacgcggccg gctgcaccaa gctgttttcc
      661 gagaagatca ccggcaccag gcgcgaccgc ccggagctgg ccaggatgct tgaccaccta
      721 cgccctggcg acgttgtgac agtgaccagg ctagaccgcc tggcccgcag cacccgcgac
      781 ctactggaca ttgccgagcg catccaggag gccggcgcgg gcctgcgtag cctggcagag
      841 ccgtgggccg acaccaccac gccggccggc cgcatggtgt tgaccgtgtt cgccggcatt
      901 gccgagttcg agcgttccct aatcatcgac cgcacccgga gcgggcgcga ggccgccaag
      961 gcccgaggcg tgaagtttgg cccccgccct accctcaccc cggcacagat cgcgcacgcc
     1021 cgcgagctga tcgaccagga aggccgcacc gtgaaagagg cggctgcact gcttggcgtg
     1081 catcgctcga ccctgtaccg cgcacttgag cgcagcgagg aagtgacgcc caccgaggcc
     1141 aggcggcgcg gtgccttccg tgaggacgca ttgaccgagg ccgacgccct ggcggccgcc
     1201 gagaatgaac gccaagagga acaagcatga aaccgcacca ggacggccag gacgaaccgt
     1261 ttttcattac cgaagagatc gaggcggaga tgatcgcggc cgggtacgtg ttcgagccgc
     1321 ccgcgcacct ctcaaccgtg cggctgcatg aaatcctggc cggtttgtct gatgccaagc
     1381 tggcggcctg gccggccagc ttggccgctg aagaaaccga gcgccgccgt ctaaaaaggt
     1441 gatgtgtatt tgagtaaaac agcttgcgtc atgcggtcgc tgcgtatatg atgcgatgag
     1501 taaataaaca aatacgcaag gggaacgcat gaaggttatc gctgtactta accagaaagg
     1561 cgggtcaggc aagacgacca tcgcaaccca tctagcccgc gccctgcaac tcgccggggc
     1621 cgatgttctg ttagtcgatt ccgatcccca gggcagtgcc cgcgattggg cggccgtgcg
     1681 ggaagatcaa ccgctaaccg ttgtcggcat cgaccgcccg acgattgacc gcgacgtgaa
     1741 ggccatcggc cggcgcgact tcgtagtgat cgacggagcg ccccaggcgg cggacttggc
     1801 tgtgtccgcg atcaaggcag ccgacttcgt gctgattccg gtgcagccaa gcccttacga
     1861 catatgggcc accgccgacc tggtggagct ggttaagcag cgcattgagg tcacggatgg
     1921 aaggctacaa gcggcctttg tcgtgtcgcg ggcgatcaaa ggcacgcgca tcggcggtga
     1981 ggttgccgag gcgctggccg ggtacgagct gcccattctt gagtcccgta tcacgcagcg
     2041 cgtgagctac ccaggcactg ccgccgccgg cacaaccgtt cttgaatcag aacccgaggg
     2101 cgacgctgcc cgcgaggtcc aggcgctggc cgctgaaatt aaatcaaaac tcatttgagt
     2161 taatgaggta aagagaaaat gagcaaaagc acaaacacgc taagtgccgg ccgtccgagc
     2221 gcacgcagca gcaaggctgc aacgttggcc agcctggcag acacgccagc catgaagcgg
     2281 gtcaactttc agttgccggc ggaggatcac accaagctga agatgtacgc ggtacgccaa
     2341 ggcaagacca ttaccgagct gctatctgaa tacatcgcgc agctaccaga gtaaatgagc
     2401 aaatgaataa atgagtagat gaattttagc ggctaaagga ggcggcatgg aaaatcaaga
     2461 acaaccaggc accgacgccg tggaatgccc catgtgtgga ggaacgggcg gttggccagg
     2521 cgtaagcggc tgggttgcct gccggccctg caatggcact ggaaccccca agcccgagga
     2581 atcggcgtga gcggtcgcaa accatccggc ccggtacaaa tcggcgcggc gctgggtgat
     2641 gacctggtgg agaagttgaa ggccgcgcag gccgcccagc ggcaacgcat cgaggcagaa
     2701 gcacgccccg gtgaatcgtg gcaagcggcc gctgatcgaa tccgcaaaga atcccggcaa
     2761 ccgccggcag ccggtgcgcc gtcgattagg aagccgccca agggcgacga gcaaccagat
     2821 tttttcgttc cgatgctcta tgacgtgggc acccgcgata gtcgcagcat catggacgtg
     2881 gccgttttcc gtctgtcgaa gcgtgaccga cgagctggcg aggtgatccg ctacgagctt
     2941 ccagacgggc acgtagaggt ttccgcaggg ccggccggca tggccagtgt gtgggattac
     3001 gacctggtac tgatggcggt ttcccatcta accgaatcca tgaaccgata ccgggaaggg
     3061 aagggagaca agcccggccg cgtgttccgt ccacacgttg cggacgtact caagttctgc
     3121 cggcgagccg atggcggaaa gcagaaagac gacctggtag aaacctgcat tcggttaaac
     3181 accacgcacg ttgccatgca gcgtacgaag aaggccaaga acggccgcct ggtgacggta
     3241 tccgagggtg aagccttgat tagccgctac aagatcgtaa agagcgaaac cgggcggccg
     3301 gagtacatcg agatcgagct agctgattgg atgtaccgcg agatcacaga aggcaagaac
     3361 ccggacgtgc tgacggttca ccccgattac tttttgatcg atcccggcat cggccgtttt
     3421 ctctaccgcc tggcacgccg cgccgcaggc aaggcagaag ccagatggtt gttcaagacg
     3481 atctacgaac gcagtggcag cgccggagag ttcaagaagt tctgtttcac cgtgcgcaag
     3541 ctgatcgggt caaatgacct gccggagtac gatttgaagg aggaggcggg gcaggctggc
     3601 ccgatcctag tcatgcgcta ccgcaacctg atcgagggcg aagcatccgc cggttcctaa
     3661 tgtacggagc agatgctagg gcaaattgcc ctagcagggg aaaaaggtcg aaaaggactc
     3721 tttcctgtgg atagcacgta cattgggaac ccaaagccgt acattgggaa ccggaacccg
     3781 tacattggga acccaaagcc gtacattggg aaccggtcac acatgtaagt gactgatata
     3841 aaagagaaaa aaggcgattt ttccgcctaa aactctttaa aacttattaa aactcttaaa
     3901 acccgcctgg cctgtgcata actgtctggc cagcgcacag ccgaagagct gcaaaaagcg
     3961 cctacccttc ggtcgctgcg ctccctacgc cccgccgctt cgcgtcggcc tatcgcggcc
     4021 gctggccgct caaaaatggc tggcctacgg ccaggcaatc taccagggcg cggacaagcc
     4081 gcgccgtcgc cactcgaccg ccggcgccca catcaaggca ccctgcctcg cgcgtttcgg
     4141 tgatgacggt gaaaacctct gacacatgca gctcccggtg acggtcacag cttgtctgta
     4201 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     4261 gggcgcagcc atgacccagt cacgtagcga tagcggagtg tatactggct taactatgcg
     4321 gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc
     4381 gtaaggagaa aataccgcat caggcgctct tccgcttcct cgctcactga ctcgctgcgc
     4441 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     4501 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     4561 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     4621 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     4681 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     4741 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     4801 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     4861 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     4921 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     4981 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     5041 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     5101 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     5161 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     5221 aacgaaaact cacgttaagg gattttggtc atgcattcta ggtgattatt tgccgactac
     5281 cttggtgatc tcgcctttca cgtagtggac aaattcttcc aactgatctg cgcgcgaggc
     5341 caagcgatct tcttcttgtc caagataagc ctgtctagct tcaagtatga cgggctgata
     5401 ctgggccggc aggcgctcca ttgcccagtc ggcagcgaca tccttcggcg cgattttgcc
     5461 ggttactgcg ctgtaccaaa tgcgggacaa cgtaagcact acatttcgct catcaccagc
     5521 ccagtcgggc ggcgagttcc atagcgttaa ggtttcattt agcgcctcaa atagatcctg
     5581 ttcaggaacc ggatcaaaga gttcctccgc cgctggacct accaaggcaa cgctatgttc
     5641 tcttgctttt gtcagcaaga tagccagatc aatgtcgatc gtggctggct cgaagatacc
     5701 tgcaagaatg tcattgcgct gccattctcc aaattgcagt tcgcgcttag ctggataacg
     5761 ccacggaatg atgtcgtcgt gcacaacaat ggtgacttct acagcgcgga gaatctcgct
     5821 ctctccaggg gaagccgaag tttccaaaag gtcgttgatc aaagctcgcc gcgttgtttc
     5881 atcaagcctt acggtcaccg taaccagcaa atcaatatca ctgtgtggct tcaggccgcc
     5941 atccactgcg gagccgtaca aatgtacggc cagcaacgtc ggttcgagat ggcgctcgat
     6001 gacgccaact acctctgata gttgagtcga tacttcggcg atcaccgctt ccctcataat
     6061 gtttaacttt gttttagggc gactgccctg ctgcgtaaca tcgttgctgc tccataacat
     6121 caaacatcga cccacggcgt aacgcgcttg ctgcttggat gcccgaggca tagactgtac
     6181 cccaaaaaaa cagtcataac aagccatgaa aaccgccact gcgccgttac caccgctgcg
     6241 ttcggtcaag gttctggacc agttgcgtga gcgcatacgc tacttgcatt acagcttacg
     6301 aaccgaacag gcttatgtcc actgggttcg tgccttcatc cgtttccacg gtgtgcgtca
     6361 cccggcaacc ttgggtagca gcgaagtcga ggcatttctg tcctggctgg aacagaactt
     6421 attatttcct tcctcttttc tacagtattt aaagataccc caagaagcta attataacaa
     6481 gacgaactcc aattcactgt tccttgcatt ctaaaacctt aaataccaga aaacagcttt
     6541 ttcaaagttg ttttcaaagt tggcgtataa catagtatcg acggagccga ttttgaaacc
     6601 gcggtgatca caggcagcaa cgctctgtca tcgttacaat caacatgcta ccctccgcga
     6661 gatcatccgt gtttcaaacc cggcagctta gttgccgttc ttccgaatag catcggtaac
     6721 atgagcaaag tctgccgcct tacaacggct ctcccgctga cgccgtcccg gactgatggg
     6781 ctgcctgtat cgagtggtga ttttgtgccg agctgccggt cggggagctg ttggctggct
     6841 ggtggcagga tatattgtgg tgtaaaca
//
