LOCUS       sequence-416785-        4096 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 4096)
  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 4096)
  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: pSEVA13-GG
FEATURES             Location/Qualifiers
     source          1..4096
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      519..1290
                     /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(1405..1491)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     1506..1528
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     RBS             1699..1707
                     /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      2524..2618
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     promoter        2691..2782
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2783..2851,2852..3643)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     terminator      3647..3686
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     oriT            3833..3941
                     /note="incP origin of transfer"
     CDS             complement(join(3952..4096,1..518))
                     /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 atgcccggaa tgccgggctg gctgggcggc tcctcgccgg ggccggtcgg tagttgctgc
       61 tcgcccggat acagggtcgg gatgcggcgc aggtcgccat gccccaacag cgattcgtcc
      121 tggtcgtcgt gatcaaccac cacggcggca ctgaacaccg acaggcgcaa ctggtcgcgg
      181 ggctggcccc acgccacgcg gtcattgacc acgtaggccg acacggtgcc ggggccgttg
      241 agcttcacga cggagatcca gcgctcggcc accaagtcct tgactgcgta ttggaccgtc
      301 cgcaaagaac gtccgatgag cttggaaagt gtcttctggc tgaccaccac ggcgttctgg
      361 tggcccatct gcgccacgag gtgatgcagc agcattgccg ccgtgggttt cctcgcaata
      421 agcccggccc acgcctcatg cgctttgcgt tccgtttgca cccagtgacc gggcttgttc
      481 ttggcttgaa tgccgatttc tctggactgc gtggccatgc ttatctccat gcggtagggg
      541 tgccgcacgg ttgcggcacc atgcgcaatc agctgcaact tttcggcagc gcgacaacaa
      601 ttatgcgttg cgtaaaagtg gcagtcaatt acagattttc tttaacctac gcaatgagct
      661 attgcggggg gtgccgcaat gagctgttgc gtacccccct tttttaagtt gttgattttt
      721 aagtctttcg catttcgccc tatatctagt tctttggtgc ccaaagaagg gcacccctgc
      781 ggggttcccc cacgccttcg gcgcggctcc ccctccggca aaaagtggcc cctccggggc
      841 ttgttgatcg actgcgcggc cttcggcctt gcccaaggtg gcgctgcccc cttggaaccc
      901 ccgcactcgc cgccgtgagg ctcggggggc aggcgggcgg gcttcgccct tcgactgccc
      961 ccactcgcat aggcttgggt cgttccaggc gcgtcaaggc caagccgctg cgcggtcgct
     1021 gcgcgagcct tgacccgcct tccacttggt gtccaaccgg caagcgaagc gcgcaggccg
     1081 caggccggag gcttttcccc agagaaaatt aaaaaaattg atggggcaag gccgcaggcc
     1141 gcgcagttgg agccggtggg tatgtggtcg aaggctgggt agccggtggg caatccctgt
     1201 ggtcaagctc gtgggcaggc gcagcctgtc catcagcttg tccagcaggg ttgtccacgg
     1261 gccgagcgaa gcgagccagc cggtggccgc tcgcggccat cgtccacata tccacgggct
     1321 ggcaagggag cgcagcgacc gcgcagggcg aagcccggag agcaagcccg tagggggggc
     1381 gcgcccagct gtctagggcg gcggatttgt cctactcagg agagcgttca ccgacaaaca
     1441 acagataaaa cgaaaggccc agtctttcga ctgagccttt cgttttattt gatgccttta
     1501 attaaagcgg ataacaattt cacacaggag gccgcctagg ccgcggccgc gcgaattcga
     1561 gctcggtacc cggggattca gagacctgcc cattgacaag gctctcgcgg ccaggtataa
     1621 ttgcacgagg gcccaagttc acttaaaaag gagatcaaca atgaaagcaa ttttcgtact
     1681 gaaacatctt aatcatgcta aggaggtttt ctaatgcgta aaggtgaaga actgttcacc
     1741 ggtgttgttc cgatcctggt tgaactggat ggtgatgtta acggccacaa attctctgtt
     1801 cgtggtgaag gtgaaggtga tgcaaccaac ggtaaactga ccctgaaatt catctgcact
     1861 accggtaaac tgccggttcc atggccgact ctggtgacta ccctgaccta tggtgttcag
     1921 tgtttttctc gttacccgga tcacatgaag cagcatgatt tcttcaaatc tgcaatgccg
     1981 gaaggttatg tacaggagcg caccatttct ttcaaagacg atggcaccta caaaacccgt
     2041 gcagaggtta aatttgaagg tgatactctg gtgaaccgta ttgaactgaa aggcattgat
     2101 ttcaaagagg acggcaacat cctgggccac aaactggaat ataacttcaa ctcccataac
     2161 gtttacatca ccgcagacaa acagaagaac ggtatcaaag ctaacttcaa aattcgccat
     2221 aacgttgaag acggtagcgt acagctggcg gaccactacc agcagaacac tccgatcggt
     2281 gatggtccgg ttctgctgcc ggataaccac tacctgtcca cccagtctaa actgtccaaa
     2341 gacccgaacg aaaagcgcga ccacatggtg ctgctggagt tcgttactgc cgcaggtatc
     2401 acgcacggca tggatgaact ctacaaataa atgactagtg gtctcgtcgc ccggcaaaac
     2461 cgggcgtgga ttgaaacgct tgcggccgcg tcgtgactgg gaaaaccctg gcgactagtc
     2521 ttggactcct gttgatagat ccagtaatga cctcagaact ccatctggat ttgttcagaa
     2581 cgctcggttg ccgccgggcg ttttttattg gtgagaatcc aggggtcccc aataattacg
     2641 atttaaatta gtagcccgcc taatgagcgg gctttttttt aattccccta tttgtttatt
     2701 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     2761 ataatattga aaaaggaaga gtatgagcat tcagcatttt cgtgtggcgc tgattccgtt
     2821 ttttgcggcg ttttgcctgc cggtgtttgc gcatccggaa accctggtga aagtgaaaga
     2881 tgcggaagat caactgggtg cgcgcgtggg ctatattgaa ctggatctga acagcggcaa
     2941 aattctggaa tcttttcgtc cggaagaacg ttttccgatg atgagcacct ttaaagtgct
     3001 gctgtgcggt gcggttctga gccgtgtgga tgcgggccag gaacaactgg gccgtcgtat
     3061 tcattatagc cagaacgatc tggtggaata tagcccggtg accgaaaaac atctgaccga
     3121 tggcatgacc gtgcgtgaac tgtgcagcgc ggcgattacc atgagcgata acaccgcggc
     3181 gaacctgctg ctgacgacca ttggcggtcc gaaagaactg accgcgtttc tgcataacat
     3241 gggcgatcat gtgacccgtc tggatcgttg ggaaccggaa ctgaacgaag cgattccgaa
     3301 cgatgaacgt gataccacca tgccggcagc aatggcgacc accctgcgta aactgctgac
     3361 gggtgagctg ctgaccctgg caagccgcca gcaactgatt gattggatgg aagcggataa
     3421 agtggcgggt ccgctgctgc gtagcgcgct gccggctggc tggtttattg cggataaaag
     3481 cggtgcgggc gaacgtggca gccgtggcat tattgcggcg ctgggcccgg atggtaaacc
     3541 gagccgtatt gtggtgattt ataccaccgg cagccaggcg acgatggatg aacgtaaccg
     3601 tcagattgcg gaaattggcg cgagcctgat taaacattgg taaaccgata caattaaagg
     3661 ctccttttgg agcctttttt tttggacgac ccttgtcctt ttccgctgca taaccctgct
     3721 tcggggtcat tatagcgatt ttttcggtat atccatcctt tttcgcacga tatacaggat
     3781 tttgccaaag ggttcgtgta gactttcctt ggtgtatcca acggcgtcag ccgggcagga
     3841 taggtgaagt aggcccaccc gcgagcgggt gttccttctt cactgtccct tattcgcacc
     3901 tggcggtgct caacgggaat cctgctctgc gaggctggcc gtaggccggc cctaccggcg
     3961 cggcagcgtt acccgtgtcg gcggctccaa cggctcgcca tcgtccagaa aacacggctc
     4021 atcgggcatc ggcaggcgct gctgcccgcg ccgttcccat tcctccgttt cggtcaaggc
     4081 tggcaggtct ggttcc
//
