LOCUS       sequence-416774-        4054 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  for cloning homology arms, with GFP as counter selection marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4054)
  AUTHORS   Lammens E-M, Volke DC, Schroven K, Voet M, Kerremans A, Lavigne R, 
            Hendrix H
  TITLE     A SEVA-based, CRISPR-Cas3-assisted genome engineering approach for 
            Pseudomonas with efficient vector curing.
  JOURNAL   Microbiol Spectr. 2023 Nov 17:e0270723. doi: 
            10.1128/spectrum.02707-23.
  PUBMED    37975669
REFERENCE   2  (bases 1 to 4054)
  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: "Microbiol 
            Spectr. 2023 Nov 17:e0270723. doi: 10.1128/spectrum.02707-23."
COMMENT     Sequence Label: pSEVA43-GG
FEATURES             Location/Qualifiers
     source          1..4054
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..23
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     RBS             194..202
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     terminator      1019..1113
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1349..2140
                     /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"
     oriT            2286..2394
                     /note="incP origin of transfer"
     CDS             complement(2405..3067)
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     rep_origin      3068..3839
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     terminator      complement(3954..4040)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 agcggataac aatttcacac aggaggccgc ctaggccgcg gccgcgcgaa ttcgagctcg
       61 gtacccgggg attcagagac ctgcccattg acaaggctct cgcggccagg tataattgca
      121 cgagggccca agttcactta aaaaggagat caacaatgaa agcaattttc gtactgaaac
      181 atcttaatca tgctaaggag gttttctaat gcgtaaaggt gaagaactgt tcaccggtgt
      241 tgttccgatc ctggttgaac tggatggtga tgttaacggc cacaaattct ctgttcgtgg
      301 tgaaggtgaa ggtgatgcaa ccaacggtaa actgaccctg aaattcatct gcactaccgg
      361 taaactgccg gttccatggc cgactctggt gactaccctg acctatggtg ttcagtgttt
      421 ttctcgttac ccggatcaca tgaagcagca tgatttcttc aaatctgcaa tgccggaagg
      481 ttatgtacag gagcgcacca tttctttcaa agacgatggc acctacaaaa cccgtgcaga
      541 ggttaaattt gaaggtgata ctctggtgaa ccgtattgaa ctgaaaggca ttgatttcaa
      601 agaggacggc aacatcctgg gccacaaact ggaatataac ttcaactccc ataacgttta
      661 catcaccgca gacaaacaga agaacggtat caaagctaac ttcaaaattc gccataacgt
      721 tgaagacggt agcgtacagc tggcggacca ctaccagcag aacactccga tcggtgatgg
      781 tccggttctg ctgccggata accactacct gtccacccag tctaaactgt ccaaagaccc
      841 gaacgaaaag cgcgaccaca tggtgctgct ggagttcgtt actgccgcag gtatcacgca
      901 cggcatggat gaactctaca aataaatgac tagtggtctc gtcgcccggc aaaaccgggc
      961 gtggattgaa acgcttgcgg ccgcgtcgtg actgggaaaa ccctggcgac tagtcttgga
     1021 ctcctgttga tagatccagt aatgacctca gaactccatc tggatttgtt cagaacgctc
     1081 ggttgccgcc gggcgttttt tattggtgag aatccagggg tccccaataa ttacgattta
     1141 cgtatttaaa tgaaccttga ccgaacgcag cggtggtaac ggcgcagtgg cggttttcat
     1201 ggcttgttat gactgttttt ttggggtaca gtctatgcct cgggcatcca agcagcaagc
     1261 gcgttacgcc gtgggtcgat gtttgatgtt atggagcagc aacgatgtta cgcagcaggg
     1321 cagtcgccct aaaacaaagt taaacatcat gagggaagcg gtgatcgccg aagtatcgac
     1381 tcaactatca gaggtagttg gcgtcatcga gcgccatctc gaaccgacgt tgctggccgt
     1441 acatttgtac ggctccgcag tggatggcgg cctgaagcca cacagtgata ttgatttgct
     1501 ggttacggtg accgtaaggc ttgatgaaac aacgcggcga gctttgatca acgacctttt
     1561 ggaaacttcg gcttcccctg gagagagcga gattctccgc gctgtagaag tcaccattgt
     1621 tgtgcacgac gacatcattc cgtggcgtta tccagctaag cgcgaactgc aatttggaga
     1681 atggcagcgc aatgacattc ttgcaggtat cttcgagcca gccacgatcg acattgatct
     1741 ggctatcttg ctgacaaaag caagagaaca tagcgttgcc ttggtaggtc cagcggcgga
     1801 ggaactcttt gatccggttc ctgaacagga tctatttgag gcgctaaatg aaaccttaac
     1861 gctatggaac tcgccgcccg actgggctgg cgatgagcga aatgtagtgc ttacgttgtc
     1921 ccgcatttgg tacagcgcag taaccggcaa aatcgcgccg aaggatgtcg ctgccgactg
     1981 ggcaatggag cgcctgccgg cccagtatca gcccgtcata cttgaagcta gacaggctta
     2041 tcttggacaa gaagaagatc gcttggcctc gcgcgcagat cagttggaag aatttgtcca
     2101 ctacgtgaaa ggcgagatca ccaaggtagt cggcaaataa gacaattgtc cttttccgct
     2161 gcataaccct gcttcggggt cattatagcg attttttcgg tatatccatc ctttttcgca
     2221 cgatatacag gattttgcca aagggttcgt gtagactttc cttggtgtat ccaacggcgt
     2281 cagccgggca ggataggtga agtaggccca cccgcgagcg ggtgttcctt cttcactgtc
     2341 ccttattcgc acctggcggt gctcaacggg aatcctgctc tgcgaggctg gccgtaggcc
     2401 ggccctaccg gcgcggcagc gttacccgtg tcggcggctc caacggctcg ccatcgtcca
     2461 gaaaacacgg ctcatcgggc atcggcaggc gctgctgccc gcgccgttcc cattcctccg
     2521 tttcggtcaa ggctggcagg tctggttcca tgcccggaat gccgggctgg ctgggcggct
     2581 cctcgccggg gccggtcggt agttgctgct cgcccggata cagggtcggg atgcggcgca
     2641 ggtcgccatg ccccaacagc gattcgtcct ggtcgtcgtg atcaaccacc acggcggcac
     2701 tgaacaccga caggcgcaac tggtcgcggg gctggcccca cgccacgcgg tcattgacca
     2761 cgtaggccga cacggtgccg gggccgttga gcttcacgac ggagatccag cgctcggcca
     2821 ccaagtcctt gactgcgtat tggaccgtcc gcaaagaacg tccgatgagc ttggaaagtg
     2881 tcttctggct gaccaccacg gcgttctggt ggcccatctg cgccacgagg tgatgcagca
     2941 gcattgccgc cgtgggtttc ctcgcaataa gcccggccca cgcctcatgc gctttgcgtt
     3001 ccgtttgcac ccagtgaccg ggcttgttct tggcttgaat gccgatttct ctggactgcg
     3061 tggccatgct tatctccatg cggtaggggt gccgcacggt tgcggcacca tgcgcaatca
     3121 gctgcaactt ttcggcagcg cgacaacaat tatgcgttgc gtaaaagtgg cagtcaatta
     3181 cagattttct ttaacctacg caatgagcta ttgcgggggg tgccgcaatg agctgttgcg
     3241 tacccccctt ttttaagttg ttgattttta agtctttcgc atttcgccct atatctagtt
     3301 ctttggtgcc caaagaaggg cacccctgcg gggttccccc acgccttcgg cgcggctccc
     3361 cctccggcaa aaagtggccc ctccggggct tgttgatcga ctgcgcggcc ttcggccttg
     3421 cccaaggtgg cgctgccccc ttggaacccc cgcactcgcc gccgtgaggc tcggggggca
     3481 ggcgggcggg cttcgccctt cgactgcccc cactcgcata ggcttgggtc gttccaggcg
     3541 cgtcaaggcc aagccgctgc gcggtcgctg cgcgagcctt gacccgcctt ccacttggtg
     3601 tccaaccggc aagcgaagcg cgcaggccgc aggccggagg cttttcccca gagaaaatta
     3661 aaaaaattga tggggcaagg ccgcaggccg cgcagttgga gccggtgggt atgtggtcga
     3721 aggctgggta gccggtgggc aatccctgtg gtcaagctcg tgggcaggcg cagcctgtcc
     3781 atcagcttgt ccagcagggt tgtccacggg ccgagcgaag cgagccagcc ggtggccgct
     3841 cgcggccatc gtccacatat ccacgggctg gcaagggagc gcagcgaccg cgcagggcga
     3901 agcccggaga gcaagcccgt agggggggcg cgcccagctg tctagggcgg cggatttgtc
     3961 ctactcagga gagcgttcac cgacaaacaa cagataaaac gaaaggccca gtctttcgac
     4021 tgagcctttc gttttatttg atgcctttaa ttaa
//
