LOCUS       sequence-415147-        7286 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GreenBraid reverse destination vector pGBZ1r - PaqCI - Eco31I - 
            spisPink- Eco31I - PaqCI.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7286)
  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 7286)
  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: pMP120
FEATURES             Location/Qualifiers
     source          1..7286
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    2..26
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             459..1274
                     /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             459..1274
                     /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     complement(532..551)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1362..1950
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1851..1870
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2104..2121
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2136..2276
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2202..2221)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2362..2384
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      2620..2814
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(2880..3953)
                     /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"
     CDS             complement(4382..5011)
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     misc_feature    6312..6336
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     promoter        6381..6415
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     RBS             6424..6435
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      7124..7195
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      7211..7238
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
ORIGIN
        1 tgtttacacc acaatatatc ctgccaccag ccagccaaca gctccccgac cggcagctcg
       61 gcacaaaatc accactcgat acaggcagcc catcagtccg ggacggcgtc agcgggagag
      121 ccgttgtaag gcggcagact ttgctcatgt taccgatgct attcggaaga acggcaacta
      181 agctgccggg tttgaaacac ggatgatctc gcggagggta gcatgttgat tgtaacgatg
      241 acagagcgtt gctgcctgtg atcaccgcgg tttcaaaatc ggctccgtcg atactatgtt
      301 atacgccaac tttgaaaaca actttgaaaa agctgttttc tggtatttaa ggttttagaa
      361 tgcaaggaac agtgaattgg agttcgtctt gttataatta gcttcttggg gtatctttaa
      421 atactgtaga aaagaggaag gaaataataa gttctgttat gagccatatt caacgggaaa
      481 cgtcttgctc aaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat
      541 gggctcgcga taatgtcggg caatcaggtg cgacaatcta ccgattgtat gggaagcccg
      601 atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg
      661 agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta
      721 tccgtactcc tgatgatgca tggttactca ccactgcgat cccagggaaa acagcattcc
      781 aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc
      841 tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc
      901 gactcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg
      961 acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat
     1021 tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg
     1081 aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg
     1141 atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt
     1201 ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg
     1261 atgagttttt ctaatcacct agaatgcatg accaaaatcc cttaacgtga gttttcgttc
     1321 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     1381 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     1441 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     1501 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     1561 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     1621 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     1681 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     1741 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     1801 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     1861 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     1921 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     1981 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     2041 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     2101 cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg
     2161 catctgtgcg gtatttcaca ccgcatatgg tgcactctca gtacaatctg ctctgatgcc
     2221 gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg gctgcgcccc
     2281 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     2341 acagacaagc tgtgaccgtc accgggagct gcatgtgtca gaggttttca ccgtcatcac
     2401 cgaaacgcgc gaggcagggt gccttgatgt gggcgccggc ggtcgagtgg cgacggcgcg
     2461 gcttgtccgc gccctggtag attgcctggc cgtaggccag ccatttttga gcggccagcg
     2521 gccgcgatag gccgacgcga agcggcgggg cgtagggagc gcagcgaccg aagggtaggc
     2581 gctttttgca gctcttcggc tgtgcgctgg ccagacagtt atgcacaggc caggcgggtt
     2641 ttaagagttt taataagttt taaagagttt taggcggaaa aatcgccttt tttctctttt
     2701 atatcagtca cttacatgtg tgaccggttc ccaatgtacg gctttgggtt cccaatgtac
     2761 gggttccggt tcccaatgta cggctttggg ttcccaatgt acgtgctatc cacaggaaag
     2821 agtccttttc gacctttttc ccctgctagg gcaatttgcc ctagcatctg ctccgtacat
     2881 taggaaccgg cggatgcttc gccctcgatc aggttgcggt agcgcatgac taggatcggg
     2941 ccagcctgcc ccgcctcctc cttcaaatcg tactccggca ggtcatttga cccgatcagc
     3001 ttgcgcacgg tgaaacagaa cttcttgaac tctccggcgc tgccactgcg ttcgtagatc
     3061 gtcttgaaca accatctggc ttctgccttg cctgcggcgc ggcgtgccag gcggtagaga
     3121 aaacggccga tgccgggatc gatcaaaaag taatcggggt gaaccgtcag cacgtccggg
     3181 ttcttgcctt ctgtgatctc gcggtacatc caatcagcta gctcgatctc gatgtactcc
     3241 ggccgcccgg tttcgctctt tacgatcttg tagcggctaa tcaaggcttc accctcggat
     3301 accgtcacca ggcggccgtt cttggccttc ttcgtacgct gcatggcaac gtgcgtggtg
     3361 tttaaccgaa tgcaggtttc taccaggtcg tctttctgct ttccgccatc ggctcgccgg
     3421 cagaacttga gtacgtccgc aacgtgtgga cggaacacgc ggccgggctt gtctcccttc
     3481 ccttcccggt atcggttcat ggattcggtt agatgggaaa ccgccatcag taccaggtcg
     3541 taatcccaca cactggccat gccggccggc cctgcggaaa cctctacgtg cccgtctgga
     3601 agctcgtagc ggatcacctc gccagctcgt cggtcacgct tcgacagacg gaaaacggcc
     3661 acgtccatga tgctgcgact atcgcgggtg cccacgtcat agagcatcgg aacgaaaaaa
     3721 tctggttgct cgtcgccctt gggcggcttc ctaatcgacg gcgcaccggc tgccggcggt
     3781 tgccgggatt ctttgcggat tcgatcagcg gccgcttgcc acgattcacc ggggcgtgct
     3841 tctgcctcga tgcgttgccg ctgggcggcc tgcgcggcct tcaacttctc caccaggtca
     3901 tcacccagcg ccgcgccgat ttgtaccggg ccggatggtt tgcgaccgct cacgccgatt
     3961 cctcgggctt gggggttcca gtgccattgc agggccggca ggcaacccag ccgcttacgc
     4021 ctggccaacc gcccgttcct ccacacatgg ggcattccac ggcgtcggtg cctggttgtt
     4081 cttgattttc catgccgcct cctttagccg ctaaaattca tctactcatt tattcatttg
     4141 ctcatttact ctggtagctg cgcgatgtat tcagatagca gctcggtaat ggtcttgcct
     4201 tggcgtaccg cgtacatctt cagcttggtg tgatcctccg ccggcaactg aaagttgacc
     4261 cgcttcatgg ctggcgtgtc tgccaggctg gccaacgttg cagccttgct gctgcgtgcg
     4321 ctcggacggc cggcacttag cgtgtttgtg cttttgctca ttttctcttt acctcattaa
     4381 ctcaaatgag ttttgattta atttcagcgg ccagcgcctg gacctcgcgg gcagcgtcgc
     4441 cctcgggttc tgattcaaga acggttgtgc cggcggcggc agtgcctggg tagctcacgc
     4501 gctgcgtgat acgggactca agaatgggca gctcgtaccc ggccagcgcc tcggcaacct
     4561 caccgccgat gcgcgtgcct ttgatcgccc gcgacacgac aaaggccgct tgtagccttc
     4621 catccgtgac ctcaatgcgc tgcttaacca gctccaccag gtcggcggtg gcccatatgt
     4681 cgtaagggct tggctgcacc ggaatcagca cgaagtcggc tgccttgatc gcggacacag
     4741 ccaagtccgc cgcctggggc gctccgtcga tcactacgaa gtcgcgccgg ccgatggcct
     4801 tcacgtcgcg gtcaatcgtc gggcggtcga tgccgacaac ggttagcggt tgatcttccc
     4861 gcacggccgc ccaatcgcgg gcactgccct ggggatcgga atcgactaac agaacatcgg
     4921 ccccggcgag ttgcagggcg cgggctagat gggttgcgat ggtcgtcttg cctgacccgc
     4981 ctttctggtt aagtacagcg ataaccttca tgcgttcccc ttgcgtattt gtttatttac
     5041 tcatcgcatc atatacgcag cgaccgcatg acgcaagctg ttttactcaa atacacatca
     5101 cctttttaga cggcggcgct cggtttcttc agcggccaag ctggccggcc aggccgccag
     5161 cttggcatca gacaaaccgg ccaggatttc atgcagccgc acggttgaga ggtgcgcggg
     5221 cggctcgaac acgtacccgg ccgcgatcat ctccgcctcg atctcttcgg taatgaaaaa
     5281 cggttcgtcc tggccgtcct ggtgcggttt catgcttgtt cctcttggcg ttcattctcg
     5341 gcggccgcca gggcgtcggc ctcggtcaat gcgtcctcac ggaaggcacc gcgccgcctg
     5401 gcctcggtgg gcgtcacttc ctcgctgcgc tcaagtgcgc ggtacagggt cgagcgatgc
     5461 acgccaagca gtgcagccgc ctctttcacg gtgcggcctt cctggtcgat cagctcgcgg
     5521 gcgtgcgcga tctgtgccgg ggtgagggta gggcgggggc caaacttcac gcctcgggcc
     5581 ttggcggcct cgcgcccgct ccgggtgcgg tcgatgatta gggaacgctc gaactcggca
     5641 atgccggcga acacggtcaa caccatgcgg ccggccggcg tggtggtgtc ggcccacggc
     5701 tctgccaggc tacgcaggcc cgcgccggcc tcctggatgc gctcggcaat gtccagtagg
     5761 tcgcgggtgc tgcgggccag gcggtctagc ctggtcactg tcacaacgtc gccagggcgt
     5821 aggtggtcaa gcatcctggc cagctccggg cggtcgcgcc tggtgccggt gatcttctcg
     5881 gaaaacagct tggtgcagcc ggccgcgtgc agttcggccc gttggttggt caagtcctgg
     5941 tcgtcggtgc tgacgcgggc atagcccagc aggccagcgg cggcgctctt gttcatggcg
     6001 taatgtctcc ggttctagtc gcaagtattc tactttatgc gactaaaaca cgcgacaaga
     6061 aaacgccagg aaaagggcag ggcggcagcc tgtcgcgtaa cttaggactt gtgcgacatg
     6121 tcgttttcag atgacggctg cactgaacgt cagaagccga ctgcactata gcagcggagg
     6181 ggttggatca aagtactttg atcccgaggg gaaccctgtg gttggcatgc acatacaaat
     6241 ggacgaacgg ataaaccttt tcacgccctt ttaaatatcc gattattcta ataaacgctc
     6301 ttttctctta ggtttacccg ccaatatatc ctgtcagaat taccatggca cctgctacaa
     6361 cctatacaga gaccctgcag tttacggcta gctcagtcct aggtatagtg ctagctacta
     6421 gagaaagagg agaaatacta aatgtcgcac tcaaaacaag cactggcaga tacgatgaaa
     6481 atgacgtggc tgatggaagg ttcggtcaac ggtcacgcct ttacgattga aggcgaaggt
     6541 accggcaaac cgtatgaagg taaacagagt ggcacctttc gtgttacgaa aggcggtccg
     6601 ctgccgtttg cgtttgatat tgtcgctccg accctgaaat acggtttcaa atgcttcatg
     6661 aaatacccgg cggatattcc ggactacttt aaactggcct tcccggaagg cctgacgtac
     6721 gatcgtaaaa ttgcgtttga agacggcggt tgtgcgaccg ccacggtcga aatgagcctg
     6781 aagggtaaca ccctggtgca taaaacgaac tttcagggcg gtaatttccc gattgatggt
     6841 ccggtgatgc aaaaacgcac cctgggctgg gaaccgacct ctgaaaaaat gacgccgtgc
     6901 gatggtatta tcaaaggcga caccatcatg tacctgatgg ttgaaggcgg taaaacgctg
     6961 aaatgtcgtt atgaaaacaa ttaccgcgcc aacaaaccag tgctgatgcc gccgagccac
     7021 tttgtggatc tgcgtctgac ccgcacgaat ctggataaag aaggtctggc gttcaaactg
     7081 gaagaatatg ctgttgcccg tgtgctggaa gtgtaataag cttcaaataa aacgaaaggc
     7141 tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctctacta
     7201 gagtcacact ggctcacctt cgggtgggcc tttctgcggg atccggtctc taggtgtcaa
     7261 tcggcaggtg ctcgagcaat tcgtta
//
