LOCUS       sequence-415150-        6548 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GreenBraid joker destination vector pGBZ1j - PaqCI - dummy - PaqCI.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6548)
  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 6548)
  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: pMP072
FEATURES             Location/Qualifiers
     source          1..6548
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    213..237
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             1538..2167
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             2596..3669
                     /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      3735..3929
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(4165..4187)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4273..4413
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4328..4347
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4428..4445)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4599..5187)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4679..4698)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5275..6090)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(5275..6090)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     5998..6017
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     misc_feature    6523..6547
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 taacgaatta ccatggcacc tgctacaacc ttctcaatgg taactttact ctttatttaa
       61 ccatacattt ttttttattt ttttcacttt gttcttcatc cactattgtt ctttgttcat
      121 cttgaacaaa agctccctcc ttctttgttc ttcatccacc attgttcttc atcaatcatt
      181 tcgctgtcat gtagcaggtg ctcgagcaat tctgacagga tatattggcg ggtaaaccta
      241 agagaaaaga gcgtttatta gaataatcgg atatttaaaa gggcgtgaaa aggtttatcc
      301 gttcgtccat ttgtatgtgc atgccaacca cagggttccc ctcgggatca aagtactttg
      361 atccaacccc tccgctgcta tagtgcagtc ggcttctgac gttcagtgca gccgtcatct
      421 gaaaacgaca tgtcgcacaa gtcctaagtt acgcgacagg ctgccgccct gcccttttcc
      481 tggcgttttc ttgtcgcgtg ttttagtcgc ataaagtaga atacttgcga ctagaaccgg
      541 agacattacg ccatgaacaa gagcgccgcc gctggcctgc tgggctatgc ccgcgtcagc
      601 accgacgacc aggacttgac caaccaacgg gccgaactgc acgcggccgg ctgcaccaag
      661 ctgttttccg agaagatcac cggcaccagg cgcgaccgcc cggagctggc caggatgctt
      721 gaccacctac gccctggcga cgttgtgaca gtgaccaggc tagaccgcct ggcccgcagc
      781 acccgcgacc tactggacat tgccgagcgc atccaggagg ccggcgcggg cctgcgtagc
      841 ctggcagagc cgtgggccga caccaccacg ccggccggcc gcatggtgtt gaccgtgttc
      901 gccggcattg ccgagttcga gcgttcccta atcatcgacc gcacccggag cgggcgcgag
      961 gccgccaagg cccgaggcgt gaagtttggc ccccgcccta ccctcacccc ggcacagatc
     1021 gcgcacgccc gcgagctgat cgaccaggaa ggccgcaccg tgaaagaggc ggctgcactg
     1081 cttggcgtgc atcgctcgac cctgtaccgc gcacttgagc gcagcgagga agtgacgccc
     1141 accgaggcca ggcggcgcgg tgccttccgt gaggacgcat tgaccgaggc cgacgccctg
     1201 gcggccgccg agaatgaacg ccaagaggaa caagcatgaa accgcaccag gacggccagg
     1261 acgaaccgtt tttcattacc gaagagatcg aggcggagat gatcgcggcc gggtacgtgt
     1321 tcgagccgcc cgcgcacctc tcaaccgtgc ggctgcatga aatcctggcc ggtttgtctg
     1381 atgccaagct ggcggcctgg ccggccagct tggccgctga agaaaccgag cgccgccgtc
     1441 taaaaaggtg atgtgtattt gagtaaaaca gcttgcgtca tgcggtcgct gcgtatatga
     1501 tgcgatgagt aaataaacaa atacgcaagg ggaacgcatg aaggttatcg ctgtacttaa
     1561 ccagaaaggc gggtcaggca agacgaccat cgcaacccat ctagcccgcg ccctgcaact
     1621 cgccggggcc gatgttctgt tagtcgattc cgatccccag ggcagtgccc gcgattgggc
     1681 ggccgtgcgg gaagatcaac cgctaaccgt tgtcggcatc gaccgcccga cgattgaccg
     1741 cgacgtgaag gccatcggcc ggcgcgactt cgtagtgatc gacggagcgc cccaggcggc
     1801 ggacttggct gtgtccgcga tcaaggcagc cgacttcgtg ctgattccgg tgcagccaag
     1861 cccttacgac atatgggcca ccgccgacct ggtggagctg gttaagcagc gcattgaggt
     1921 cacggatgga aggctacaag cggcctttgt cgtgtcgcgg gcgatcaaag gcacgcgcat
     1981 cggcggtgag gttgccgagg cgctggccgg gtacgagctg cccattcttg agtcccgtat
     2041 cacgcagcgc gtgagctacc caggcactgc cgccgccggc acaaccgttc ttgaatcaga
     2101 acccgagggc gacgctgccc gcgaggtcca ggcgctggcc gctgaaatta aatcaaaact
     2161 catttgagtt aatgaggtaa agagaaaatg agcaaaagca caaacacgct aagtgccggc
     2221 cgtccgagcg cacgcagcag caaggctgca acgttggcca gcctggcaga cacgccagcc
     2281 atgaagcggg tcaactttca gttgccggcg gaggatcaca ccaagctgaa gatgtacgcg
     2341 gtacgccaag gcaagaccat taccgagctg ctatctgaat acatcgcgca gctaccagag
     2401 taaatgagca aatgaataaa tgagtagatg aattttagcg gctaaaggag gcggcatgga
     2461 aaatcaagaa caaccaggca ccgacgccgt ggaatgcccc atgtgtggag gaacgggcgg
     2521 ttggccaggc gtaagcggct gggttgcctg ccggccctgc aatggcactg gaacccccaa
     2581 gcccgaggaa tcggcgtgag cggtcgcaaa ccatccggcc cggtacaaat cggcgcggcg
     2641 ctgggtgatg acctggtgga gaagttgaag gccgcgcagg ccgcccagcg gcaacgcatc
     2701 gaggcagaag cacgccccgg tgaatcgtgg caagcggccg ctgatcgaat ccgcaaagaa
     2761 tcccggcaac cgccggcagc cggtgcgccg tcgattagga agccgcccaa gggcgacgag
     2821 caaccagatt ttttcgttcc gatgctctat gacgtgggca cccgcgatag tcgcagcatc
     2881 atggacgtgg ccgttttccg tctgtcgaag cgtgaccgac gagctggcga ggtgatccgc
     2941 tacgagcttc cagacgggca cgtagaggtt tccgcagggc cggccggcat ggccagtgtg
     3001 tgggattacg acctggtact gatggcggtt tcccatctaa ccgaatccat gaaccgatac
     3061 cgggaaggga agggagacaa gcccggccgc gtgttccgtc cacacgttgc ggacgtactc
     3121 aagttctgcc ggcgagccga tggcggaaag cagaaagacg acctggtaga aacctgcatt
     3181 cggttaaaca ccacgcacgt tgccatgcag cgtacgaaga aggccaagaa cggccgcctg
     3241 gtgacggtat ccgagggtga agccttgatt agccgctaca agatcgtaaa gagcgaaacc
     3301 gggcggccgg agtacatcga gatcgagcta gctgattgga tgtaccgcga gatcacagaa
     3361 ggcaagaacc cggacgtgct gacggttcac cccgattact ttttgatcga tcccggcatc
     3421 ggccgttttc tctaccgcct ggcacgccgc gccgcaggca aggcagaagc cagatggttg
     3481 ttcaagacga tctacgaacg cagtggcagc gccggagagt tcaagaagtt ctgtttcacc
     3541 gtgcgcaagc tgatcgggtc aaatgacctg ccggagtacg atttgaagga ggaggcgggg
     3601 caggctggcc cgatcctagt catgcgctac cgcaacctga tcgagggcga agcatccgcc
     3661 ggttcctaat gtacggagca gatgctaggg caaattgccc tagcagggga aaaaggtcga
     3721 aaaggactct ttcctgtgga tagcacgtac attgggaacc caaagccgta cattgggaac
     3781 cggaacccgt acattgggaa cccaaagccg tacattggga accggtcaca catgtaagtg
     3841 actgatataa aagagaaaaa aggcgatttt tccgcctaaa actctttaaa acttattaaa
     3901 actcttaaaa cccgcctggc ctgtgcataa ctgtctggcc agcgcacagc cgaagagctg
     3961 caaaaagcgc ctacccttcg gtcgctgcgc tccctacgcc ccgccgcttc gcgtcggcct
     4021 atcgcggccg ctggccgctc aaaaatggct ggcctacggc caggcaatct accagggcgc
     4081 ggacaagccg cgccgtcgcc actcgaccgc cggcgcccac atcaaggcac cctgcctcgc
     4141 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggtga cggtcacagc
     4201 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     4261 cgggtgtcgg ggcgcagcca tgacccagtc acgtagcgat agcggagtgt atactggctt
     4321 aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg
     4381 cacagatgcg taaggagaaa ataccgcatc aggcgctctt ccgcttcctc gctcactgac
     4441 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     4501 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     4561 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     4621 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     4681 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     4741 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     4801 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     4861 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     4921 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     4981 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     5041 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     5101 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     5161 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     5221 gctcagtgga acgaaaactc acgttaaggg attttggtca tgcattctag gtgattagaa
     5281 aaactcatcg agcatcaaat gaaactgcaa tttattcata tcaggattat caataccata
     5341 tttttgaaaa agccgtttct gtaatgaagg agaaaactca ccgaggcagt tccataggat
     5401 ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca acatcaatac aacctattaa
     5461 tttcccctcg tcaaaaataa ggttatcaag tgagaaatca ccatgagtga cgactgaatc
     5521 cggtgagaat ggcaaaagtt tatgcatttc tttccagact tgttcaacag gccagccatt
     5581 acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg
     5641 agcgagtcga aatacgcgat cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa
     5701 ccggcgcagg aacactgcca gcgcatcaac aatattttca cctgaatcag gatattcttc
     5761 taatacctgg aatgctgttt tccctgggat cgcagtggtg agtaaccatg catcatcagg
     5821 agtacggata aaatgcttga tggtcggaag aggcataaat tccgtcagcc agtttagtct
     5881 gaccatctca tctgtaacat cattggcaac gctacctttg ccatgtttca gaaacaactc
     5941 tggcgcatcg ggcttcccat acaatcggta gattgtcgca cctgattgcc cgacattatc
     6001 gcgagcccat ttatacccat ataaatcagc atccatgttg gaatttaatc gcggccttga
     6061 gcaagacgtt tcccgttgaa tatggctcat aacagaactt attatttcct tcctcttttc
     6121 tacagtattt aaagataccc caagaagcta attataacaa gacgaactcc aattcactgt
     6181 tccttgcatt ctaaaacctt aaataccaga aaacagcttt ttcaaagttg ttttcaaagt
     6241 tggcgtataa catagtatcg acggagccga ttttgaaacc gcggtgatca caggcagcaa
     6301 cgctctgtca tcgttacaat caacatgcta ccctccgcga gatcatccgt gtttcaaacc
     6361 cggcagctta gttgccgttc ttccgaatag catcggtaac atgagcaaag tctgccgcct
     6421 tacaacggct ctcccgctga cgccgtcccg gactgatggg ctgcctgtat cgagtggtga
     6481 ttttgtgccg agctgccggt cggggagctg ttggctggct ggtggcagga tatattgtgg
     6541 tgtaaaca
//
