LOCUS       sequence-415334-        7653 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GreenBraid destination vector pGBY2 - Eco31I - PaqCI -sfGFP- PaqCI -
            Eco31I.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7653)
  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 7653)
  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: pMP119
FEATURES             Location/Qualifiers
     source          1..7653
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        49..83
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     RBS             92..103
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      834..905
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      921..948
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    989..1013
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             2314..2943
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             3372..4445
                     /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      4511..4705
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(4941..4963)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    5049..5189
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     5104..5123
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5204..5221)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5375..5963)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5455..5474)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6051..6842)
                     /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    7628..7652
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 taacggatca ccatggggtc tcagtcaacc tatcggcagg tgctgcagtt tacggctagc
       61 tcagtcctag gtatagtgct agctactaga gaaagaggag aaatactaaa tgcgtaaagg
      121 cgaagaactg ttcacgggcg tagttccgat tctggtcgag ctggacggcg atgtgaacgg
      181 tcataagttt agcgttcgcg gtgaaggtga gggcgacgcg accaacggca aactgaccct
      241 gaagttcatc tgcaccaccg gtaaactgcc ggtgccttgg ccgaccttgg tgacgacgtt
      301 gacgtatggc gtgcagtgtt ttgcgcgtta tccggaccac atgaaacaac acgatttctt
      361 caaatctgcg atgccggagg gttacgtcca ggagcgtacc atttccttca aggatgatgg
      421 ctactacaaa actcgcgcag aggttaagtt tgaaggtgac acgctggtca atcgtatcga
      481 attgaagggt atcgacttta aagaggatgg taacattctg ggccataaac tggagtataa
      541 cttcaacagc cataatgttt acattacggc agacaagcaa aagaacggca tcaaggccaa
      601 tttcaagatt cgccacaatg ttgaggacgg tagcgtccaa ctggccgacc attaccagca
      661 gaacacccca attggtgacg gtccggtttt gctgccggat aatcactatc tgagcaccca
      721 aagcgtgctg agcaaagatc cgaacgaaaa acgtgatcac atggtcctgc tggaatttgt
      781 gaccgctgcg ggcatcaccc acggtatgga cgagctgtat aagtaatgag cttcaaataa
      841 aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg
      901 ctctctacta gagtcacact ggctcacctt cgggtgggcc tttctgcggg atcccacctg
      961 ctacagtatt gagaccctcg agagatcctg acaggatata ttggcgggta aacctaagag
     1021 aaaagagcgt ttattagaat aatcggatat ttaaaagggc gtgaaaaggt ttatccgttc
     1081 gtccatttgt atgtgcatgc caaccacagg gttcccctcg ggatcaaagt actttgatcc
     1141 aacccctccg ctgctatagt gcagtcggct tctgacgttc agtgcagccg tcatctgaaa
     1201 acgacatgtc gcacaagtcc taagttacgc gacaggctgc cgccctgccc ttttcctggc
     1261 gttttcttgt cgcgtgtttt agtcgcataa agtagaatac ttgcgactag aaccggagac
     1321 attacgccat gaacaagagc gccgccgctg gcctgctggg ctatgcccgc gtcagcaccg
     1381 acgaccagga cttgaccaac caacgggccg aactgcacgc ggccggctgc accaagctgt
     1441 tttccgagaa gatcaccggc accaggcgcg accgcccgga gctggccagg atgcttgacc
     1501 acctacgccc tggcgacgtt gtgacagtga ccaggctaga ccgcctggcc cgcagcaccc
     1561 gcgacctact ggacattgcc gagcgcatcc aggaggccgg cgcgggcctg cgtagcctgg
     1621 cagagccgtg ggccgacacc accacgccgg ccggccgcat ggtgttgacc gtgttcgccg
     1681 gcattgccga gttcgagcgt tccctaatca tcgaccgcac ccggagcggg cgcgaggccg
     1741 ccaaggcccg aggcgtgaag tttggccccc gccctaccct caccccggca cagatcgcgc
     1801 acgcccgcga gctgatcgac caggaaggcc gcaccgtgaa agaggcggct gcactgcttg
     1861 gcgtgcatcg ctcgaccctg taccgcgcac ttgagcgcag cgaggaagtg acgcccaccg
     1921 aggccaggcg gcgcggtgcc ttccgtgagg acgcattgac cgaggccgac gccctggcgg
     1981 ccgccgagaa tgaacgccaa gaggaacaag catgaaaccg caccaggacg gccaggacga
     2041 accgtttttc attaccgaag agatcgaggc ggagatgatc gcggccgggt acgtgttcga
     2101 gccgcccgcg cacctctcaa ccgtgcggct gcatgaaatc ctggccggtt tgtctgatgc
     2161 caagctggcg gcctggccgg ccagcttggc cgctgaagaa accgagcgcc gccgtctaaa
     2221 aaggtgatgt gtatttgagt aaaacagctt gcgtcatgcg gtcgctgcgt atatgatgcg
     2281 atgagtaaat aaacaaatac gcaaggggaa cgcatgaagg ttatcgctgt acttaaccag
     2341 aaaggcgggt caggcaagac gaccatcgca acccatctag cccgcgccct gcaactcgcc
     2401 ggggccgatg ttctgttagt cgattccgat ccccagggca gtgcccgcga ttgggcggcc
     2461 gtgcgggaag atcaaccgct aaccgttgtc ggcatcgacc gcccgacgat tgaccgcgac
     2521 gtgaaggcca tcggccggcg cgacttcgta gtgatcgacg gagcgcccca ggcggcggac
     2581 ttggctgtgt ccgcgatcaa ggcagccgac ttcgtgctga ttccggtgca gccaagccct
     2641 tacgacatat gggccaccgc cgacctggtg gagctggtta agcagcgcat tgaggtcacg
     2701 gatggaaggc tacaagcggc ctttgtcgtg tcgcgggcga tcaaaggcac gcgcatcggc
     2761 ggtgaggttg ccgaggcgct ggccgggtac gagctgccca ttcttgagtc ccgtatcacg
     2821 cagcgcgtga gctacccagg cactgccgcc gccggcacaa ccgttcttga atcagaaccc
     2881 gagggcgacg ctgcccgcga ggtccaggcg ctggccgctg aaattaaatc aaaactcatt
     2941 tgagttaatg aggtaaagag aaaatgagca aaagcacaaa cacgctaagt gccggccgtc
     3001 cgagcgcacg cagcagcaag gctgcaacgt tggccagcct ggcagacacg ccagccatga
     3061 agcgggtcaa ctttcagttg ccggcggagg atcacaccaa gctgaagatg tacgcggtac
     3121 gccaaggcaa gaccattacc gagctgctat ctgaatacat cgcgcagcta ccagagtaaa
     3181 tgagcaaatg aataaatgag tagatgaatt ttagcggcta aaggaggcgg catggaaaat
     3241 caagaacaac caggcaccga cgccgtggaa tgccccatgt gtggaggaac gggcggttgg
     3301 ccaggcgtaa gcggctgggt tgcctgccgg ccctgcaatg gcactggaac ccccaagccc
     3361 gaggaatcgg cgtgagcggt cgcaaaccat ccggcccggt acaaatcggc gcggcgctgg
     3421 gtgatgacct ggtggagaag ttgaaggccg cgcaggccgc ccagcggcaa cgcatcgagg
     3481 cagaagcacg ccccggtgaa tcgtggcaag cggccgctga tcgaatccgc aaagaatccc
     3541 ggcaaccgcc ggcagccggt gcgccgtcga ttaggaagcc gcccaagggc gacgagcaac
     3601 cagatttttt cgttccgatg ctctatgacg tgggcacccg cgatagtcgc agcatcatgg
     3661 acgtggccgt tttccgtctg tcgaagcgtg accgacgagc tggcgaggtg atccgctacg
     3721 agcttccaga cgggcacgta gaggtttccg cagggccggc cggcatggcc agtgtgtggg
     3781 attacgacct ggtactgatg gcggtttccc atctaaccga atccatgaac cgataccggg
     3841 aagggaaggg agacaagccc ggccgcgtgt tccgtccaca cgttgcggac gtactcaagt
     3901 tctgccggcg agccgatggc ggaaagcaga aagacgacct ggtagaaacc tgcattcggt
     3961 taaacaccac gcacgttgcc atgcagcgta cgaagaaggc caagaacggc cgcctggtga
     4021 cggtatccga gggtgaagcc ttgattagcc gctacaagat cgtaaagagc gaaaccgggc
     4081 ggccggagta catcgagatc gagctagctg attggatgta ccgcgagatc acagaaggca
     4141 agaacccgga cgtgctgacg gttcaccccg attacttttt gatcgatccc ggcatcggcc
     4201 gttttctcta ccgcctggca cgccgcgccg caggcaaggc agaagccaga tggttgttca
     4261 agacgatcta cgaacgcagt ggcagcgccg gagagttcaa gaagttctgt ttcaccgtgc
     4321 gcaagctgat cgggtcaaat gacctgccgg agtacgattt gaaggaggag gcggggcagg
     4381 ctggcccgat cctagtcatg cgctaccgca acctgatcga gggcgaagca tccgccggtt
     4441 cctaatgtac ggagcagatg ctagggcaaa ttgccctagc aggggaaaaa ggtcgaaaag
     4501 gactctttcc tgtggatagc acgtacattg ggaacccaaa gccgtacatt gggaaccgga
     4561 acccgtacat tgggaaccca aagccgtaca ttgggaaccg gtcacacatg taagtgactg
     4621 atataaaaga gaaaaaaggc gatttttccg cctaaaactc tttaaaactt attaaaactc
     4681 ttaaaacccg cctggcctgt gcataactgt ctggccagcg cacagccgaa gagctgcaaa
     4741 aagcgcctac ccttcggtcg ctgcgctccc tacgccccgc cgcttcgcgt cggcctatcg
     4801 cggccgctgg ccgctcaaaa atggctggcc tacggccagg caatctacca gggcgcggac
     4861 aagccgcgcc gtcgccactc gaccgccggc gcccacatca aggcaccctg cctcgcgcgt
     4921 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggtgacggt cacagcttgt
     4981 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg
     5041 tgtcggggcg cagccatgac ccagtcacgt agcgatagcg gagtgtatac tggcttaact
     5101 atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca
     5161 gatgcgtaag gagaaaatac cgcatcaggc gctcttccgc ttcctcgctc actgactcgc
     5221 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     5281 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     5341 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     5401 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     5461 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     5521 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     5581 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     5641 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     5701 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     5761 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
     5821 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     5881 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     5941 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     6001 agtggaacga aaactcacgt taagggattt tggtcatgca ttctaggtga ttatttgccg
     6061 actaccttgg tgatctcgcc tttcacgtag tggacaaatt cttccaactg atctgcgcgc
     6121 gaggccaagc gatcttcttc ttgtccaaga taagcctgtc tagcttcaag tatgacgggc
     6181 tgatactggg ccggcaggcg ctccattgcc cagtcggcag cgacatcctt cggcgcgatt
     6241 ttgccggtta ctgcgctgta ccaaatgcgg gacaacgtaa gcactacatt tcgctcatca
     6301 ccagcccagt cgggcggcga gttccatagc gttaaggttt catttagcgc ctcaaataga
     6361 tcctgttcag gaaccggatc aaagagttcc tccgccgctg gacctaccaa ggcaacgcta
     6421 tgttctcttg cttttgtcag caagatagcc agatcaatgt cgatcgtggc tggctcgaag
     6481 atacctgcaa gaatgtcatt gcgctgccat tctccaaatt gcagttcgcg cttagctgga
     6541 taacgccacg gaatgatgtc gtcgtgcaca acaatggtga cttctacagc gcggagaatc
     6601 tcgctctctc caggggaagc cgaagtttcc aaaaggtcgt tgatcaaagc tcgccgcgtt
     6661 gtttcatcaa gccttacggt caccgtaacc agcaaatcaa tatcactgtg tggcttcagg
     6721 ccgccatcca ctgcggagcc gtacaaatgt acggccagca acgtcggttc gagatggcgc
     6781 tcgatgacgc caactacctc tgatagttga gtcgatactt cggcgatcac cgcttccctc
     6841 ataatgttta actttgtttt agggcgactg ccctgctgcg taacatcgtt gctgctccat
     6901 aacatcaaac atcgacccac ggcgtaacgc gcttgctgct tggatgcccg aggcatagac
     6961 tgtaccccaa aaaaacagtc ataacaagcc atgaaaaccg ccactgcgcc gttaccaccg
     7021 ctgcgttcgg tcaaggttct ggaccagttg cgtgagcgca tacgctactt gcattacagc
     7081 ttacgaaccg aacaggctta tgtccactgg gttcgtgcct tcatccgttt ccacggtgtg
     7141 cgtcacccgg caaccttggg tagcagcgaa gtcgaggcat ttctgtcctg gctggaacag
     7201 aacttattat ttccttcctc ttttctacag tatttaaaga taccccaaga agctaattat
     7261 aacaagacga actccaattc actgttcctt gcattctaaa accttaaata ccagaaaaca
     7321 gctttttcaa agttgttttc aaagttggcg tataacatag tatcgacgga gccgattttg
     7381 aaaccgcggt gatcacaggc agcaacgctc tgtcatcgtt acaatcaaca tgctaccctc
     7441 cgcgagatca tccgtgtttc aaacccggca gcttagttgc cgttcttccg aatagcatcg
     7501 gtaacatgag caaagtctgc cgccttacaa cggctctccc gctgacgccg tcccggactg
     7561 atgggctgcc tgtatcgagt ggtgattttg tgccgagctg ccggtcgggg agctgttggc
     7621 tggctggtgg caggatatat tgtggtgtaa aca
//
