LOCUS       sequence-415320-        6869 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GreenBraid joker destination vector pGBY2j - Eco31I - dummy - 
            Eco31I.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6869)
  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 6869)
  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: pMP075
FEATURES             Location/Qualifiers
     source          1..6869
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    205..229
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             1530..2159
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             2588..3661
                     /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      3727..3921
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(4157..4179)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4265..4405
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4320..4339
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4420..4437)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4591..5179)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4671..4690)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5267..6058)
                     /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    6844..6868
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 taacggatca ccatggggtc tcagtcagga gtctcaatgg taactttact ctttatttaa
       61 ccatacattt ttttttattt ttttcacttt gttcttcatc cactattgtt ctttgttcat
      121 cttgaacaaa agctccctcc ttctttgttc ttcatccacc attgttcttc atcaatcatt
      181 tgtattgaga ccctcgagag atcctgacag gatatattgg cgggtaaacc taagagaaaa
      241 gagcgtttat tagaataatc ggatatttaa aagggcgtga aaaggtttat ccgttcgtcc
      301 atttgtatgt gcatgccaac cacagggttc ccctcgggat caaagtactt tgatccaacc
      361 cctccgctgc tatagtgcag tcggcttctg acgttcagtg cagccgtcat ctgaaaacga
      421 catgtcgcac aagtcctaag ttacgcgaca ggctgccgcc ctgccctttt cctggcgttt
      481 tcttgtcgcg tgttttagtc gcataaagta gaatacttgc gactagaacc ggagacatta
      541 cgccatgaac aagagcgccg ccgctggcct gctgggctat gcccgcgtca gcaccgacga
      601 ccaggacttg accaaccaac gggccgaact gcacgcggcc ggctgcacca agctgttttc
      661 cgagaagatc accggcacca ggcgcgaccg cccggagctg gccaggatgc ttgaccacct
      721 acgccctggc gacgttgtga cagtgaccag gctagaccgc ctggcccgca gcacccgcga
      781 cctactggac attgccgagc gcatccagga ggccggcgcg ggcctgcgta gcctggcaga
      841 gccgtgggcc gacaccacca cgccggccgg ccgcatggtg ttgaccgtgt tcgccggcat
      901 tgccgagttc gagcgttccc taatcatcga ccgcacccgg agcgggcgcg aggccgccaa
      961 ggcccgaggc gtgaagtttg gcccccgccc taccctcacc ccggcacaga tcgcgcacgc
     1021 ccgcgagctg atcgaccagg aaggccgcac cgtgaaagag gcggctgcac tgcttggcgt
     1081 gcatcgctcg accctgtacc gcgcacttga gcgcagcgag gaagtgacgc ccaccgaggc
     1141 caggcggcgc ggtgccttcc gtgaggacgc attgaccgag gccgacgccc tggcggccgc
     1201 cgagaatgaa cgccaagagg aacaagcatg aaaccgcacc aggacggcca ggacgaaccg
     1261 tttttcatta ccgaagagat cgaggcggag atgatcgcgg ccgggtacgt gttcgagccg
     1321 cccgcgcacc tctcaaccgt gcggctgcat gaaatcctgg ccggtttgtc tgatgccaag
     1381 ctggcggcct ggccggccag cttggccgct gaagaaaccg agcgccgccg tctaaaaagg
     1441 tgatgtgtat ttgagtaaaa cagcttgcgt catgcggtcg ctgcgtatat gatgcgatga
     1501 gtaaataaac aaatacgcaa ggggaacgca tgaaggttat cgctgtactt aaccagaaag
     1561 gcgggtcagg caagacgacc atcgcaaccc atctagcccg cgccctgcaa ctcgccgggg
     1621 ccgatgttct gttagtcgat tccgatcccc agggcagtgc ccgcgattgg gcggccgtgc
     1681 gggaagatca accgctaacc gttgtcggca tcgaccgccc gacgattgac cgcgacgtga
     1741 aggccatcgg ccggcgcgac ttcgtagtga tcgacggagc gccccaggcg gcggacttgg
     1801 ctgtgtccgc gatcaaggca gccgacttcg tgctgattcc ggtgcagcca agcccttacg
     1861 acatatgggc caccgccgac ctggtggagc tggttaagca gcgcattgag gtcacggatg
     1921 gaaggctaca agcggccttt gtcgtgtcgc gggcgatcaa aggcacgcgc atcggcggtg
     1981 aggttgccga ggcgctggcc gggtacgagc tgcccattct tgagtcccgt atcacgcagc
     2041 gcgtgagcta cccaggcact gccgccgccg gcacaaccgt tcttgaatca gaacccgagg
     2101 gcgacgctgc ccgcgaggtc caggcgctgg ccgctgaaat taaatcaaaa ctcatttgag
     2161 ttaatgaggt aaagagaaaa tgagcaaaag cacaaacacg ctaagtgccg gccgtccgag
     2221 cgcacgcagc agcaaggctg caacgttggc cagcctggca gacacgccag ccatgaagcg
     2281 ggtcaacttt cagttgccgg cggaggatca caccaagctg aagatgtacg cggtacgcca
     2341 aggcaagacc attaccgagc tgctatctga atacatcgcg cagctaccag agtaaatgag
     2401 caaatgaata aatgagtaga tgaattttag cggctaaagg aggcggcatg gaaaatcaag
     2461 aacaaccagg caccgacgcc gtggaatgcc ccatgtgtgg aggaacgggc ggttggccag
     2521 gcgtaagcgg ctgggttgcc tgccggccct gcaatggcac tggaaccccc aagcccgagg
     2581 aatcggcgtg agcggtcgca aaccatccgg cccggtacaa atcggcgcgg cgctgggtga
     2641 tgacctggtg gagaagttga aggccgcgca ggccgcccag cggcaacgca tcgaggcaga
     2701 agcacgcccc ggtgaatcgt ggcaagcggc cgctgatcga atccgcaaag aatcccggca
     2761 accgccggca gccggtgcgc cgtcgattag gaagccgccc aagggcgacg agcaaccaga
     2821 ttttttcgtt ccgatgctct atgacgtggg cacccgcgat agtcgcagca tcatggacgt
     2881 ggccgttttc cgtctgtcga agcgtgaccg acgagctggc gaggtgatcc gctacgagct
     2941 tccagacggg cacgtagagg tttccgcagg gccggccggc atggccagtg tgtgggatta
     3001 cgacctggta ctgatggcgg tttcccatct aaccgaatcc atgaaccgat accgggaagg
     3061 gaagggagac aagcccggcc gcgtgttccg tccacacgtt gcggacgtac tcaagttctg
     3121 ccggcgagcc gatggcggaa agcagaaaga cgacctggta gaaacctgca ttcggttaaa
     3181 caccacgcac gttgccatgc agcgtacgaa gaaggccaag aacggccgcc tggtgacggt
     3241 atccgagggt gaagccttga ttagccgcta caagatcgta aagagcgaaa ccgggcggcc
     3301 ggagtacatc gagatcgagc tagctgattg gatgtaccgc gagatcacag aaggcaagaa
     3361 cccggacgtg ctgacggttc accccgatta ctttttgatc gatcccggca tcggccgttt
     3421 tctctaccgc ctggcacgcc gcgccgcagg caaggcagaa gccagatggt tgttcaagac
     3481 gatctacgaa cgcagtggca gcgccggaga gttcaagaag ttctgtttca ccgtgcgcaa
     3541 gctgatcggg tcaaatgacc tgccggagta cgatttgaag gaggaggcgg ggcaggctgg
     3601 cccgatccta gtcatgcgct accgcaacct gatcgagggc gaagcatccg ccggttccta
     3661 atgtacggag cagatgctag ggcaaattgc cctagcaggg gaaaaaggtc gaaaaggact
     3721 ctttcctgtg gatagcacgt acattgggaa cccaaagccg tacattggga accggaaccc
     3781 gtacattggg aacccaaagc cgtacattgg gaaccggtca cacatgtaag tgactgatat
     3841 aaaagagaaa aaaggcgatt tttccgccta aaactcttta aaacttatta aaactcttaa
     3901 aacccgcctg gcctgtgcat aactgtctgg ccagcgcaca gccgaagagc tgcaaaaagc
     3961 gcctaccctt cggtcgctgc gctccctacg ccccgccgct tcgcgtcggc ctatcgcggc
     4021 cgctggccgc tcaaaaatgg ctggcctacg gccaggcaat ctaccagggc gcggacaagc
     4081 cgcgccgtcg ccactcgacc gccggcgccc acatcaaggc accctgcctc gcgcgtttcg
     4141 gtgatgacgg tgaaaacctc tgacacatgc agctcccggt gacggtcaca gcttgtctgt
     4201 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
     4261 ggggcgcagc catgacccag tcacgtagcg atagcggagt gtatactggc ttaactatgc
     4321 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg
     4381 cgtaaggaga aaataccgca tcaggcgctc ttccgcttcc tcgctcactg actcgctgcg
     4441 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     4501 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     4561 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     4621 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     4681 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     4741 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     4801 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     4861 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     4921 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     4981 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     5041 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     5101 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     5161 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     5221 gaacgaaaac tcacgttaag ggattttggt catgcattct aggtgattat ttgccgacta
     5281 ccttggtgat ctcgcctttc acgtagtgga caaattcttc caactgatct gcgcgcgagg
     5341 ccaagcgatc ttcttcttgt ccaagataag cctgtctagc ttcaagtatg acgggctgat
     5401 actgggccgg caggcgctcc attgcccagt cggcagcgac atccttcggc gcgattttgc
     5461 cggttactgc gctgtaccaa atgcgggaca acgtaagcac tacatttcgc tcatcaccag
     5521 cccagtcggg cggcgagttc catagcgtta aggtttcatt tagcgcctca aatagatcct
     5581 gttcaggaac cggatcaaag agttcctccg ccgctggacc taccaaggca acgctatgtt
     5641 ctcttgcttt tgtcagcaag atagccagat caatgtcgat cgtggctggc tcgaagatac
     5701 ctgcaagaat gtcattgcgc tgccattctc caaattgcag ttcgcgctta gctggataac
     5761 gccacggaat gatgtcgtcg tgcacaacaa tggtgacttc tacagcgcgg agaatctcgc
     5821 tctctccagg ggaagccgaa gtttccaaaa ggtcgttgat caaagctcgc cgcgttgttt
     5881 catcaagcct tacggtcacc gtaaccagca aatcaatatc actgtgtggc ttcaggccgc
     5941 catccactgc ggagccgtac aaatgtacgg ccagcaacgt cggttcgaga tggcgctcga
     6001 tgacgccaac tacctctgat agttgagtcg atacttcggc gatcaccgct tccctcataa
     6061 tgtttaactt tgttttaggg cgactgccct gctgcgtaac atcgttgctg ctccataaca
     6121 tcaaacatcg acccacggcg taacgcgctt gctgcttgga tgcccgaggc atagactgta
     6181 ccccaaaaaa acagtcataa caagccatga aaaccgccac tgcgccgtta ccaccgctgc
     6241 gttcggtcaa ggttctggac cagttgcgtg agcgcatacg ctacttgcat tacagcttac
     6301 gaaccgaaca ggcttatgtc cactgggttc gtgccttcat ccgtttccac ggtgtgcgtc
     6361 acccggcaac cttgggtagc agcgaagtcg aggcatttct gtcctggctg gaacagaact
     6421 tattatttcc ttcctctttt ctacagtatt taaagatacc ccaagaagct aattataaca
     6481 agacgaactc caattcactg ttccttgcat tctaaaacct taaataccag aaaacagctt
     6541 tttcaaagtt gttttcaaag ttggcgtata acatagtatc gacggagccg attttgaaac
     6601 cgcggtgatc acaggcagca acgctctgtc atcgttacaa tcaacatgct accctccgcg
     6661 agatcatccg tgtttcaaac ccggcagctt agttgccgtt cttccgaata gcatcggtaa
     6721 catgagcaaa gtctgccgcc ttacaacggc tctcccgctg acgccgtccc ggactgatgg
     6781 gctgcctgta tcgagtggtg attttgtgcc gagctgccgg tcggggagct gttggctggc
     6841 tggtggcagg atatattgtg gtgtaaaca
//
