LOCUS       sequence-416781-        3995 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 3995)
  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 3995)
  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: pSEVA23-GG
FEATURES             Location/Qualifiers
     source          1..3995
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      515..1286
                     /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(1401..1487)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     1502..1524
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     RBS             1695..1703
                     /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      2520..2614
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(2735..2754)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2746..3561
                     /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"
     oriT            3728..3836
                     /note="incP origin of transfer"
     CDS             complement(join(3847..3995,1..514))
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
ORIGIN
        1 ccggaatgcc gggctggctg ggcggctcct cgccggggcc ggtcggtagt tgctgctcgc
       61 ccggatacag ggtcgggatg cggcgcaggt cgccatgccc caacagcgat tcgtcctggt
      121 cgtcgtgatc aaccaccacg gcggcactga acaccgacag gcgcaactgg tcgcggggct
      181 ggccccacgc cacgcggtca ttgaccacgt aggccgacac ggtgccgggg ccgttgagct
      241 tcacgacgga gatccagcgc tcggccacca agtccttgac tgcgtattgg accgtccgca
      301 aagaacgtcc gatgagcttg gaaagtgtct tctggctgac caccacggcg ttctggtggc
      361 ccatctgcgc cacgaggtga tgcagcagca ttgccgccgt gggtttcctc gcaataagcc
      421 cggcccacgc ctcatgcgct ttgcgttccg tttgcaccca gtgaccgggc ttgttcttgg
      481 cttgaatgcc gatttctctg gactgcgtgg ccatgcttat ctccatgcgg taggggtgcc
      541 gcacggttgc ggcaccatgc gcaatcagct gcaacttttc ggcagcgcga caacaattat
      601 gcgttgcgta aaagtggcag tcaattacag attttcttta acctacgcaa tgagctattg
      661 cggggggtgc cgcaatgagc tgttgcgtac cccccttttt taagttgttg atttttaagt
      721 ctttcgcatt tcgccctata tctagttctt tggtgcccaa agaagggcac ccctgcgggg
      781 ttcccccacg ccttcggcgc ggctccccct ccggcaaaaa gtggcccctc cggggcttgt
      841 tgatcgactg cgcggccttc ggccttgccc aaggtggcgc tgcccccttg gaacccccgc
      901 actcgccgcc gtgaggctcg gggggcaggc gggcgggctt cgcccttcga ctgcccccac
      961 tcgcataggc ttgggtcgtt ccaggcgcgt caaggccaag ccgctgcgcg gtcgctgcgc
     1021 gagccttgac ccgccttcca cttggtgtcc aaccggcaag cgaagcgcgc aggccgcagg
     1081 ccggaggctt ttccccagag aaaattaaaa aaattgatgg ggcaaggccg caggccgcgc
     1141 agttggagcc ggtgggtatg tggtcgaagg ctgggtagcc ggtgggcaat ccctgtggtc
     1201 aagctcgtgg gcaggcgcag cctgtccatc agcttgtcca gcagggttgt ccacgggccg
     1261 agcgaagcga gccagccggt ggccgctcgc ggccatcgtc cacatatcca cgggctggca
     1321 agggagcgca gcgaccgcgc agggcgaagc ccggagagca agcccgtagg gggggcgcgc
     1381 ccagctgtct agggcggcgg atttgtccta ctcaggagag cgttcaccga caaacaacag
     1441 ataaaacgaa aggcccagtc tttcgactga gcctttcgtt ttatttgatg cctttaatta
     1501 aagcggataa caatttcaca caggaggccg cctaggccgc ggccgcgcga attcgagctc
     1561 ggtacccggg gattcagaga cctgcccatt gacaaggctc tcgcggccag gtataattgc
     1621 acgagggccc aagttcactt aaaaaggaga tcaacaatga aagcaatttt cgtactgaaa
     1681 catcttaatc atgctaagga ggttttctaa tgcgtaaagg tgaagaactg ttcaccggtg
     1741 ttgttccgat cctggttgaa ctggatggtg atgttaacgg ccacaaattc tctgttcgtg
     1801 gtgaaggtga aggtgatgca accaacggta aactgaccct gaaattcatc tgcactaccg
     1861 gtaaactgcc ggttccatgg ccgactctgg tgactaccct gacctatggt gttcagtgtt
     1921 tttctcgtta cccggatcac atgaagcagc atgatttctt caaatctgca atgccggaag
     1981 gttatgtaca ggagcgcacc atttctttca aagacgatgg cacctacaaa acccgtgcag
     2041 aggttaaatt tgaaggtgat actctggtga accgtattga actgaaaggc attgatttca
     2101 aagaggacgg caacatcctg ggccacaaac tggaatataa cttcaactcc cataacgttt
     2161 acatcaccgc agacaaacag aagaacggta tcaaagctaa cttcaaaatt cgccataacg
     2221 ttgaagacgg tagcgtacag ctggcggacc actaccagca gaacactccg atcggtgatg
     2281 gtccggttct gctgccggat aaccactacc tgtccaccca gtctaaactg tccaaagacc
     2341 cgaacgaaaa gcgcgaccac atggtgctgc tggagttcgt tactgccgca ggtatcacgc
     2401 acggcatgga tgaactctac aaataaatga ctagtggtct cgtcgcccgg caaaaccggg
     2461 cgtggattga aacgcttgcg gccgcgtcgt gactgggaaa accctggcga ctagtcttgg
     2521 actcctgttg atagatccag taatgacctc agaactccat ctggatttgt tcagaacgct
     2581 cggttgccgc cgggcgtttt ttattggtga gaatccaggg gtccccaata attacgattt
     2641 aaatttgtgt ctcaaaatct ctgatgttac attgcacaag ataaaaatat atcatcatga
     2701 acaataaaac tgtctgctta cataaacagt aatacaaggg gtgttatgag ccatattcag
     2761 cgtgaaacga gctgtagccg tccgcgtctg aacagcaaca tggatgcgga tctgtatggc
     2821 tataaatggg cgcgtgataa cgtgggtcag agcggcgcga ccatttatcg tctgtatggc
     2881 aaaccggatg cgccggaact gtttctgaaa catggcaaag gcagcgtggc gaacgatgtg
     2941 accgatgaaa tggtgcgtct gaactggctg accgaattta tgccgctgcc gaccattaaa
     3001 cattttattc gcaccccgga tgatgcgtgg ctgctgacca ccgcgattcc gggcaaaacc
     3061 gcgtttcagg tgctggaaga atatccggat agcggcgaaa acattgtgga tgcgctggcc
     3121 gtgtttctgc gtcgtctgca tagcattccg gtgtgcaact gcccgtttaa cagcgatcgt
     3181 gtgtttcgtc tggcccaggc gcagagccgt atgaacaacg gcctggtgga tgcgagcgat
     3241 tttgatgatg aacgtaacgg ctggccggtg gaacaggtgt ggaaagaaat gcataaactg
     3301 ctgccgttta gcccggatag cgtggtgacc cacggcgatt ttagcctgga taacctgatt
     3361 ttcgatgaag gcaaactgat tggctgcatt gatgtgggcc gtgtgggcat tgcggatcgt
     3421 tatcaggatc tggccattct gtggaactgc ctgggcgaat ttagcccgag cctgcaaaaa
     3481 cgtctgtttc agaaatatgg cattgataat ccggatatga acaaactgca atttcatctg
     3541 atgctggatg aatttttcta ataattaatt ggaccgcggt ccgcgcgttg tccttttccg
     3601 ctgcataacc ctgcttcggg gtcattatag cgattttttc ggtatatcca tcctttttcg
     3661 cacgatatac aggattttgc caaagggttc gtgtagactt tccttggtgt atccaacggc
     3721 gtcagccggg caggataggt gaagtaggcc cacccgcgag cgggtgttcc ttcttcactg
     3781 tcccttattc gcacctggcg gtgctcaacg ggaatcctgc tctgcgaggc tggccgtagg
     3841 ccggccctac cggcgcggca gcgttacccg tgtcggcggc tccaacggct cgccatcgtc
     3901 cagaaaacac ggctcatcgg gcatcggcag gcgctgctgc ccgcgccgtt cccattcctc
     3961 cgtttcggtc aaggctggca ggtctggttc catgc
//
