LOCUS       sequence-415335-        7657 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GreenBraid reverse destination vector pGBY2r - Eco31I - PaqCI 
            -sfGFP- PaqCI - Eco31I.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7657)
  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 7657)
  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: pMP123
FEATURES             Location/Qualifiers
     source          1..7657
                     /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    993..1017
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             2318..2947
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             3376..4449
                     /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      4515..4709
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(4945..4967)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    5053..5193
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     5108..5127
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5208..5225)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5379..5967)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5459..5478)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6055..6846)
                     /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    7632..7656
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 taacggatca ccatggggtc tcagtcaata catcggcagg 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 ctgcaaggtg tattgagacc ctcgagagat cctgacagga tatattggcg ggtaaaccta
     1021 agagaaaaga gcgtttatta gaataatcgg atatttaaaa gggcgtgaaa aggtttatcc
     1081 gttcgtccat ttgtatgtgc atgccaacca cagggttccc ctcgggatca aagtactttg
     1141 atccaacccc tccgctgcta tagtgcagtc ggcttctgac gttcagtgca gccgtcatct
     1201 gaaaacgaca tgtcgcacaa gtcctaagtt acgcgacagg ctgccgccct gcccttttcc
     1261 tggcgttttc ttgtcgcgtg ttttagtcgc ataaagtaga atacttgcga ctagaaccgg
     1321 agacattacg ccatgaacaa gagcgccgcc gctggcctgc tgggctatgc ccgcgtcagc
     1381 accgacgacc aggacttgac caaccaacgg gccgaactgc acgcggccgg ctgcaccaag
     1441 ctgttttccg agaagatcac cggcaccagg cgcgaccgcc cggagctggc caggatgctt
     1501 gaccacctac gccctggcga cgttgtgaca gtgaccaggc tagaccgcct ggcccgcagc
     1561 acccgcgacc tactggacat tgccgagcgc atccaggagg ccggcgcggg cctgcgtagc
     1621 ctggcagagc cgtgggccga caccaccacg ccggccggcc gcatggtgtt gaccgtgttc
     1681 gccggcattg ccgagttcga gcgttcccta atcatcgacc gcacccggag cgggcgcgag
     1741 gccgccaagg cccgaggcgt gaagtttggc ccccgcccta ccctcacccc ggcacagatc
     1801 gcgcacgccc gcgagctgat cgaccaggaa ggccgcaccg tgaaagaggc ggctgcactg
     1861 cttggcgtgc atcgctcgac cctgtaccgc gcacttgagc gcagcgagga agtgacgccc
     1921 accgaggcca ggcggcgcgg tgccttccgt gaggacgcat tgaccgaggc cgacgccctg
     1981 gcggccgccg agaatgaacg ccaagaggaa caagcatgaa accgcaccag gacggccagg
     2041 acgaaccgtt tttcattacc gaagagatcg aggcggagat gatcgcggcc gggtacgtgt
     2101 tcgagccgcc cgcgcacctc tcaaccgtgc ggctgcatga aatcctggcc ggtttgtctg
     2161 atgccaagct ggcggcctgg ccggccagct tggccgctga agaaaccgag cgccgccgtc
     2221 taaaaaggtg atgtgtattt gagtaaaaca gcttgcgtca tgcggtcgct gcgtatatga
     2281 tgcgatgagt aaataaacaa atacgcaagg ggaacgcatg aaggttatcg ctgtacttaa
     2341 ccagaaaggc gggtcaggca agacgaccat cgcaacccat ctagcccgcg ccctgcaact
     2401 cgccggggcc gatgttctgt tagtcgattc cgatccccag ggcagtgccc gcgattgggc
     2461 ggccgtgcgg gaagatcaac cgctaaccgt tgtcggcatc gaccgcccga cgattgaccg
     2521 cgacgtgaag gccatcggcc ggcgcgactt cgtagtgatc gacggagcgc cccaggcggc
     2581 ggacttggct gtgtccgcga tcaaggcagc cgacttcgtg ctgattccgg tgcagccaag
     2641 cccttacgac atatgggcca ccgccgacct ggtggagctg gttaagcagc gcattgaggt
     2701 cacggatgga aggctacaag cggcctttgt cgtgtcgcgg gcgatcaaag gcacgcgcat
     2761 cggcggtgag gttgccgagg cgctggccgg gtacgagctg cccattcttg agtcccgtat
     2821 cacgcagcgc gtgagctacc caggcactgc cgccgccggc acaaccgttc ttgaatcaga
     2881 acccgagggc gacgctgccc gcgaggtcca ggcgctggcc gctgaaatta aatcaaaact
     2941 catttgagtt aatgaggtaa agagaaaatg agcaaaagca caaacacgct aagtgccggc
     3001 cgtccgagcg cacgcagcag caaggctgca acgttggcca gcctggcaga cacgccagcc
     3061 atgaagcggg tcaactttca gttgccggcg gaggatcaca ccaagctgaa gatgtacgcg
     3121 gtacgccaag gcaagaccat taccgagctg ctatctgaat acatcgcgca gctaccagag
     3181 taaatgagca aatgaataaa tgagtagatg aattttagcg gctaaaggag gcggcatgga
     3241 aaatcaagaa caaccaggca ccgacgccgt ggaatgcccc atgtgtggag gaacgggcgg
     3301 ttggccaggc gtaagcggct gggttgcctg ccggccctgc aatggcactg gaacccccaa
     3361 gcccgaggaa tcggcgtgag cggtcgcaaa ccatccggcc cggtacaaat cggcgcggcg
     3421 ctgggtgatg acctggtgga gaagttgaag gccgcgcagg ccgcccagcg gcaacgcatc
     3481 gaggcagaag cacgccccgg tgaatcgtgg caagcggccg ctgatcgaat ccgcaaagaa
     3541 tcccggcaac cgccggcagc cggtgcgccg tcgattagga agccgcccaa gggcgacgag
     3601 caaccagatt ttttcgttcc gatgctctat gacgtgggca cccgcgatag tcgcagcatc
     3661 atggacgtgg ccgttttccg tctgtcgaag cgtgaccgac gagctggcga ggtgatccgc
     3721 tacgagcttc cagacgggca cgtagaggtt tccgcagggc cggccggcat ggccagtgtg
     3781 tgggattacg acctggtact gatggcggtt tcccatctaa ccgaatccat gaaccgatac
     3841 cgggaaggga agggagacaa gcccggccgc gtgttccgtc cacacgttgc ggacgtactc
     3901 aagttctgcc ggcgagccga tggcggaaag cagaaagacg acctggtaga aacctgcatt
     3961 cggttaaaca ccacgcacgt tgccatgcag cgtacgaaga aggccaagaa cggccgcctg
     4021 gtgacggtat ccgagggtga agccttgatt agccgctaca agatcgtaaa gagcgaaacc
     4081 gggcggccgg agtacatcga gatcgagcta gctgattgga tgtaccgcga gatcacagaa
     4141 ggcaagaacc cggacgtgct gacggttcac cccgattact ttttgatcga tcccggcatc
     4201 ggccgttttc tctaccgcct ggcacgccgc gccgcaggca aggcagaagc cagatggttg
     4261 ttcaagacga tctacgaacg cagtggcagc gccggagagt tcaagaagtt ctgtttcacc
     4321 gtgcgcaagc tgatcgggtc aaatgacctg ccggagtacg atttgaagga ggaggcgggg
     4381 caggctggcc cgatcctagt catgcgctac cgcaacctga tcgagggcga agcatccgcc
     4441 ggttcctaat gtacggagca gatgctaggg caaattgccc tagcagggga aaaaggtcga
     4501 aaaggactct ttcctgtgga tagcacgtac attgggaacc caaagccgta cattgggaac
     4561 cggaacccgt acattgggaa cccaaagccg tacattggga accggtcaca catgtaagtg
     4621 actgatataa aagagaaaaa aggcgatttt tccgcctaaa actctttaaa acttattaaa
     4681 actcttaaaa cccgcctggc ctgtgcataa ctgtctggcc agcgcacagc cgaagagctg
     4741 caaaaagcgc ctacccttcg gtcgctgcgc tccctacgcc ccgccgcttc gcgtcggcct
     4801 atcgcggccg ctggccgctc aaaaatggct ggcctacggc caggcaatct accagggcgc
     4861 ggacaagccg cgccgtcgcc actcgaccgc cggcgcccac atcaaggcac cctgcctcgc
     4921 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggtga cggtcacagc
     4981 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     5041 cgggtgtcgg ggcgcagcca tgacccagtc acgtagcgat agcggagtgt atactggctt
     5101 aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg
     5161 cacagatgcg taaggagaaa ataccgcatc aggcgctctt ccgcttcctc gctcactgac
     5221 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     5281 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     5341 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     5401 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     5461 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     5521 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     5581 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     5641 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     5701 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     5761 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     5821 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     5881 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     5941 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     6001 gctcagtgga acgaaaactc acgttaaggg attttggtca tgcattctag gtgattattt
     6061 gccgactacc ttggtgatct cgcctttcac gtagtggaca aattcttcca actgatctgc
     6121 gcgcgaggcc aagcgatctt cttcttgtcc aagataagcc tgtctagctt caagtatgac
     6181 gggctgatac tgggccggca ggcgctccat tgcccagtcg gcagcgacat ccttcggcgc
     6241 gattttgccg gttactgcgc tgtaccaaat gcgggacaac gtaagcacta catttcgctc
     6301 atcaccagcc cagtcgggcg gcgagttcca tagcgttaag gtttcattta gcgcctcaaa
     6361 tagatcctgt tcaggaaccg gatcaaagag ttcctccgcc gctggaccta ccaaggcaac
     6421 gctatgttct cttgcttttg tcagcaagat agccagatca atgtcgatcg tggctggctc
     6481 gaagatacct gcaagaatgt cattgcgctg ccattctcca aattgcagtt cgcgcttagc
     6541 tggataacgc cacggaatga tgtcgtcgtg cacaacaatg gtgacttcta cagcgcggag
     6601 aatctcgctc tctccagggg aagccgaagt ttccaaaagg tcgttgatca aagctcgccg
     6661 cgttgtttca tcaagcctta cggtcaccgt aaccagcaaa tcaatatcac tgtgtggctt
     6721 caggccgcca tccactgcgg agccgtacaa atgtacggcc agcaacgtcg gttcgagatg
     6781 gcgctcgatg acgccaacta cctctgatag ttgagtcgat acttcggcga tcaccgcttc
     6841 cctcataatg tttaactttg ttttagggcg actgccctgc tgcgtaacat cgttgctgct
     6901 ccataacatc aaacatcgac ccacggcgta acgcgcttgc tgcttggatg cccgaggcat
     6961 agactgtacc ccaaaaaaac agtcataaca agccatgaaa accgccactg cgccgttacc
     7021 accgctgcgt tcggtcaagg ttctggacca gttgcgtgag cgcatacgct acttgcatta
     7081 cagcttacga accgaacagg cttatgtcca ctgggttcgt gccttcatcc gtttccacgg
     7141 tgtgcgtcac ccggcaacct tgggtagcag cgaagtcgag gcatttctgt cctggctgga
     7201 acagaactta ttatttcctt cctcttttct acagtattta aagatacccc aagaagctaa
     7261 ttataacaag acgaactcca attcactgtt ccttgcattc taaaacctta aataccagaa
     7321 aacagctttt tcaaagttgt tttcaaagtt ggcgtataac atagtatcga cggagccgat
     7381 tttgaaaccg cggtgatcac aggcagcaac gctctgtcat cgttacaatc aacatgctac
     7441 cctccgcgag atcatccgtg tttcaaaccc ggcagcttag ttgccgttct tccgaatagc
     7501 atcggtaaca tgagcaaagt ctgccgcctt acaacggctc tcccgctgac gccgtcccgg
     7561 actgatgggc tgcctgtatc gagtggtgat tttgtgccga gctgccggtc ggggagctgt
     7621 tggctggctg gtggcaggat atattgtggt gtaaaca
//
