LOCUS       sequence-416773-        4048 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 4048)
  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 4048)
  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: pSEVA83-GG
FEATURES             Location/Qualifiers
     source          1..4048
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     9..31
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     RBS             202..210
                     /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      1027..1121
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1328..2128
                     /codon_start=1
                     /gene="aac(3)IV"
                     /product="aminoglycoside 3-N-acetyltransferase type IV     
                               "
                     /label=ApmR
                     /note="confers resistance to apramycin (Paget and Davies, 
                     1996)"
                     /translation="MSSAVECNVVQYEWRKAELIGQLLNLGVTPGGVLLVHSSFRSVRP
                     LEDGPLGLIEALRAALGPGGTLVMPSWSGLDDEPFDPATSPVTPDLGVVSDTFWRLPNV
                     KRSAHPFAFAAAGPQAEQIISDPLPLPPHSPASPVARVHELDGQVLLLGVGHDANTTLH
                     LAELMAKVPYGVPRHCTILQDGKLVRVDYLENDHCCERFALADRWLKEKSLQKEGPVGH
                     AFARLIRSRDIVATALGQLGRDPLIFLHPPEAGCEECDAARQSIG"
     oriT            2288..2396
                     /note="incP origin of transfer"
     CDS             complement(2407..3069)
                     /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      3070..3841
                     /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(3956..4042)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ttaattaaag cggataacaa tttcacacag gaggccgcct aggccgcggc cgcgcgaatt
       61 cgagctcggt acccggggat tcagagacct gcccattgac aaggctctcg cggccaggta
      121 taattgcacg agggcccaag ttcacttaaa aaggagatca acaatgaaag caattttcgt
      181 actgaaacat cttaatcatg ctaaggaggt tttctaatgc gtaaaggtga agaactgttc
      241 accggtgttg ttccgatcct ggttgaactg gatggtgatg ttaacggcca caaattctct
      301 gttcgtggtg aaggtgaagg tgatgcaacc aacggtaaac tgaccctgaa attcatctgc
      361 actaccggta aactgccggt tccatggccg actctggtga ctaccctgac ctatggtgtt
      421 cagtgttttt ctcgttaccc ggatcacatg aagcagcatg atttcttcaa atctgcaatg
      481 ccggaaggtt atgtacagga gcgcaccatt tctttcaaag acgatggcac ctacaaaacc
      541 cgtgcagagg ttaaatttga aggtgatact ctggtgaacc gtattgaact gaaaggcatt
      601 gatttcaaag aggacggcaa catcctgggc cacaaactgg aatataactt caactcccat
      661 aacgtttaca tcaccgcaga caaacagaag aacggtatca aagctaactt caaaattcgc
      721 cataacgttg aagacggtag cgtacagctg gcggaccact accagcagaa cactccgatc
      781 ggtgatggtc cggttctgct gccggataac cactacctgt ccacccagtc taaactgtcc
      841 aaagacccga acgaaaagcg cgaccacatg gtgctgctgg agttcgttac tgccgcaggt
      901 atcacgcacg gcatggatga actctacaaa taaatgacta gtggtctcgt cgcccggcaa
      961 aaccgggcgt ggattgaaac gcttgcggcc gcgtcgtgac tgggaaaacc ctggcgacta
     1021 gtcttggact cctgttgata gatccagtaa tgacctcaga actccatctg gatttgttca
     1081 gaacgctcgg ttgccgccgg gcgtttttta ttggtgagaa tccaggggtc cccaataatt
     1141 acgatttaaa tctcacggta actgatgccg tatttgcagt accagcgtac ggcccacaga
     1201 atgatgtcac gctgaaaatg ccggcctttg aatgggttca tgtgcagctc catcagcaaa
     1261 aggggatgat aagtttatca ccaccgacta tttgcaacag tgccgttgat cgtgctatga
     1321 tcgagtgatg tcatcagcgg tggagtgcaa tgtcgtgcaa tacgaatggc gaaaagccga
     1381 gctgatcggt cagcttctca accttggggt tacccccggc ggtgtgctgc tggtccacag
     1441 ctccttccgt agcgtccggc ccctcgaaga tgggccactt ggactgatcg aggccctgcg
     1501 tgctgcgctg ggtccgggag ggacgctcgt gatgccctcg tggtcaggtc tggacgacga
     1561 gccgttcgat cctgccacgt cgcccgttac accggacctt ggagttgtct ctgacacatt
     1621 ctggcgcctg ccaaatgtaa agcgcagcgc ccatccattt gcctttgcgg cagcggggcc
     1681 acaggcagag cagatcatct ctgatccatt gcccctgcca cctcactcgc ctgcaagccc
     1741 ggtcgcccgt gtccatgaac tcgatgggca ggtacttctc ctcggcgtgg gacacgatgc
     1801 caacacgacg ctgcatcttg ccgagttgat ggcaaaggtt ccctatgggg tgccgagaca
     1861 ctgcaccatt cttcaggatg gcaagttggt tcgcgtcgat tatctcgaga atgaccactg
     1921 ctgtgagcgc tttgccttgg cggacaggtg gctcaaggag aagagccttc agaaggaagg
     1981 tccagtcggt catgcctttg ctcggttgat ccgctcccgc gacattgtgg cgacagccct
     2041 gggtcaactg ggccgagatc cgttgatctt cctgcatccg ccagaggcgg gatgcgaaga
     2101 atgcgatgcc gctcgccagt cgattggcta agaccgcggt ccgcgcgttg tccttttccg
     2161 ctgcataacc ctgcttcggg gtcattatag cgattttttc ggtatatcca tcctttttcg
     2221 cacgatatac aggattttgc caaagggttc gtgtagactt tccttggtgt atccaacggc
     2281 gtcagccggg caggataggt gaagtaggcc cacccgcgag cgggtgttcc ttcttcactg
     2341 tcccttattc gcacctggcg gtgctcaacg ggaatcctgc tctgcgaggc tggccgtagg
     2401 ccggccctac cggcgcggca gcgttacccg tgtcggcggc tccaacggct cgccatcgtc
     2461 cagaaaacac ggctcatcgg gcatcggcag gcgctgctgc ccgcgccgtt cccattcctc
     2521 cgtttcggtc aaggctggca ggtctggttc catgcccgga atgccgggct ggctgggcgg
     2581 ctcctcgccg gggccggtcg gtagttgctg ctcgcccgga tacagggtcg ggatgcggcg
     2641 caggtcgcca tgccccaaca gcgattcgtc ctggtcgtcg tgatcaacca ccacggcggc
     2701 actgaacacc gacaggcgca actggtcgcg gggctggccc cacgccacgc ggtcattgac
     2761 cacgtaggcc gacacggtgc cggggccgtt gagcttcacg acggagatcc agcgctcggc
     2821 caccaagtcc ttgactgcgt attggaccgt ccgcaaagaa cgtccgatga gcttggaaag
     2881 tgtcttctgg ctgaccacca cggcgttctg gtggcccatc tgcgccacga ggtgatgcag
     2941 cagcattgcc gccgtgggtt tcctcgcaat aagcccggcc cacgcctcat gcgctttgcg
     3001 ttccgtttgc acccagtgac cgggcttgtt cttggcttga atgccgattt ctctggactg
     3061 cgtggccatg cttatctcca tgcggtaggg gtgccgcacg gttgcggcac catgcgcaat
     3121 cagctgcaac ttttcggcag cgcgacaaca attatgcgtt gcgtaaaagt ggcagtcaat
     3181 tacagatttt ctttaaccta cgcaatgagc tattgcgggg ggtgccgcaa tgagctgttg
     3241 cgtacccccc ttttttaagt tgttgatttt taagtctttc gcatttcgcc ctatatctag
     3301 ttctttggtg cccaaagaag ggcacccctg cggggttccc ccacgccttc ggcgcggctc
     3361 cccctccggc aaaaagtggc ccctccgggg cttgttgatc gactgcgcgg ccttcggcct
     3421 tgcccaaggt ggcgctgccc ccttggaacc cccgcactcg ccgccgtgag gctcgggggg
     3481 caggcgggcg ggcttcgccc ttcgactgcc cccactcgca taggcttggg tcgttccagg
     3541 cgcgtcaagg ccaagccgct gcgcggtcgc tgcgcgagcc ttgacccgcc ttccacttgg
     3601 tgtccaaccg gcaagcgaag cgcgcaggcc gcaggccgga ggcttttccc cagagaaaat
     3661 taaaaaaatt gatggggcaa ggccgcaggc cgcgcagttg gagccggtgg gtatgtggtc
     3721 gaaggctggg tagccggtgg gcaatccctg tggtcaagct cgtgggcagg cgcagcctgt
     3781 ccatcagctt gtccagcagg gttgtccacg ggccgagcga agcgagccag ccggtggccg
     3841 ctcgcggcca tcgtccacat atccacgggc tggcaaggga gcgcagcgac cgcgcagggc
     3901 gaagcccgga gagcaagccc gtaggggggg cgcgcccagc tgtctagggc ggcggatttg
     3961 tcctactcag gagagcgttc accgacaaac aacagataaa acgaaaggcc cagtctttcg
     4021 actgagcctt tcgttttatt tgatgcct
//
