LOCUS       sequence-416776-        3873 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 3873)
  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 3873)
  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: pSEVA63-GG
FEATURES             Location/Qualifiers
     source          1..3873
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     5..27
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     RBS             198..206
                     /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      1023..1117
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     promoter        join(1148..1153,1154..1170,1171..1176)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             1365..1898
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     oriT            2109..2217
                     /note="incP origin of transfer"
     CDS             complement(2228..2890)
                     /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      2891..3662
                     /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(3777..3863)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ttaaagcgga taacaatttc acacaggagg ccgcctaggc cgcggccgcg cgaattcgag
       61 ctcggtaccc ggggattcag agacctgccc attgacaagg ctctcgcggc caggtataat
      121 tgcacgaggg cccaagttca cttaaaaagg agatcaacaa tgaaagcaat tttcgtactg
      181 aaacatctta atcatgctaa ggaggttttc taatgcgtaa aggtgaagaa ctgttcaccg
      241 gtgttgttcc gatcctggtt gaactggatg gtgatgttaa cggccacaaa ttctctgttc
      301 gtggtgaagg tgaaggtgat gcaaccaacg gtaaactgac cctgaaattc atctgcacta
      361 ccggtaaact gccggttcca tggccgactc tggtgactac cctgacctat ggtgttcagt
      421 gtttttctcg ttacccggat cacatgaagc agcatgattt cttcaaatct gcaatgccgg
      481 aaggttatgt acaggagcgc accatttctt tcaaagacga tggcacctac aaaacccgtg
      541 cagaggttaa atttgaaggt gatactctgg tgaaccgtat tgaactgaaa ggcattgatt
      601 tcaaagagga cggcaacatc ctgggccaca aactggaata taacttcaac tcccataacg
      661 tttacatcac cgcagacaaa cagaagaacg gtatcaaagc taacttcaaa attcgccata
      721 acgttgaaga cggtagcgta cagctggcgg accactacca gcagaacact ccgatcggtg
      781 atggtccggt tctgctgccg gataaccact acctgtccac ccagtctaaa ctgtccaaag
      841 acccgaacga aaagcgcgac cacatggtgc tgctggagtt cgttactgcc gcaggtatca
      901 cgcacggcat ggatgaactc tacaaataaa tgactagtgg tctcgtcgcc cggcaaaacc
      961 gggcgtggat tgaaacgctt gcggccgcgt cgtgactggg aaaaccctgg cgactagtct
     1021 tggactcctg ttgatagatc cagtaatgac ctcagaactc catctggatt tgttcagaac
     1081 gctcggttgc cgccgggcgt tttttattgg tgagaatcca ggggtcccca ataattacga
     1141 tttaaatttg acataagcct gttcggttcg taaactgtaa tgcaagtagc gtatgcgctc
     1201 acgcaactgg tccagaacct tgaccgaacg cagcggtggt aacggcgcag tggcggtttt
     1261 catggcttgt tatgactgtt tttttgtaca gcctatgcct cgggcatcca agcagcaagc
     1321 gcgttacgcc gtgggtcgat gtttgatgtt atggagcagc aacgatgtta cgcagcagca
     1381 acgatgttac gcagcagggc agtcgcccta aaacaaagtt aggtggctca agtatgggca
     1441 tcattcgcac atgtaggctc ggccctgacc aagtcaaatc catgcgggct gctcttgatc
     1501 ttttcggtcg tgagttcgga gacgtagcca cctactccca acatcagccg gactccgatt
     1561 acctcgggaa cttgctccgt agtaagacat tcatcgcgct tgctgccttc gaccaagaag
     1621 cggttgttgg cgctctcgcg gcttacgttc tgcccaagtt tgagcagccg cgtagtgaga
     1681 tctatatcta tgatctcgca gtctccggag agcaccggag gcagggcatt gccaccgcgc
     1741 tcatcaatct cctcaagcat gaggccaacg cgcttggtgc ttatgtgatc tacgtgcaag
     1801 cagattacgg tgacgatccc gcagtggctc tctatacaaa gttgggcata cgggaagaag
     1861 tgatgcactt tgatatcgac ccaagtaccg ccacctaaca attcgttcaa gccgagatcg
     1921 gcttcccggc cgcggagttg ttcggtaaat tggacaacgg tccgcgcgtt gtccttttcc
     1981 gctgcataac cctgcttcgg ggtcattata gcgatttttt cggtatatcc atcctttttc
     2041 gcacgatata caggattttg ccaaagggtt cgtgtagact ttccttggtg tatccaacgg
     2101 cgtcagccgg gcaggatagg tgaagtaggc ccacccgcga gcgggtgttc cttcttcact
     2161 gtcccttatt cgcacctggc ggtgctcaac gggaatcctg ctctgcgagg ctggccgtag
     2221 gccggcccta ccggcgcggc agcgttaccc gtgtcggcgg ctccaacggc tcgccatcgt
     2281 ccagaaaaca cggctcatcg ggcatcggca ggcgctgctg cccgcgccgt tcccattcct
     2341 ccgtttcggt caaggctggc aggtctggtt ccatgcccgg aatgccgggc tggctgggcg
     2401 gctcctcgcc ggggccggtc ggtagttgct gctcgcccgg atacagggtc gggatgcggc
     2461 gcaggtcgcc atgccccaac agcgattcgt cctggtcgtc gtgatcaacc accacggcgg
     2521 cactgaacac cgacaggcgc aactggtcgc ggggctggcc ccacgccacg cggtcattga
     2581 ccacgtaggc cgacacggtg ccggggccgt tgagcttcac gacggagatc cagcgctcgg
     2641 ccaccaagtc cttgactgcg tattggaccg tccgcaaaga acgtccgatg agcttggaaa
     2701 gtgtcttctg gctgaccacc acggcgttct ggtggcccat ctgcgccacg aggtgatgca
     2761 gcagcattgc cgccgtgggt ttcctcgcaa taagcccggc ccacgcctca tgcgctttgc
     2821 gttccgtttg cacccagtga ccgggcttgt tcttggcttg aatgccgatt tctctggact
     2881 gcgtggccat gcttatctcc atgcggtagg ggtgccgcac ggttgcggca ccatgcgcaa
     2941 tcagctgcaa cttttcggca gcgcgacaac aattatgcgt tgcgtaaaag tggcagtcaa
     3001 ttacagattt tctttaacct acgcaatgag ctattgcggg gggtgccgca atgagctgtt
     3061 gcgtaccccc cttttttaag ttgttgattt ttaagtcttt cgcatttcgc cctatatcta
     3121 gttctttggt gcccaaagaa gggcacccct gcggggttcc cccacgcctt cggcgcggct
     3181 ccccctccgg caaaaagtgg cccctccggg gcttgttgat cgactgcgcg gccttcggcc
     3241 ttgcccaagg tggcgctgcc cccttggaac ccccgcactc gccgccgtga ggctcggggg
     3301 gcaggcgggc gggcttcgcc cttcgactgc ccccactcgc ataggcttgg gtcgttccag
     3361 gcgcgtcaag gccaagccgc tgcgcggtcg ctgcgcgagc cttgacccgc cttccacttg
     3421 gtgtccaacc ggcaagcgaa gcgcgcaggc cgcaggccgg aggcttttcc ccagagaaaa
     3481 ttaaaaaaat tgatggggca aggccgcagg ccgcgcagtt ggagccggtg ggtatgtggt
     3541 cgaaggctgg gtagccggtg ggcaatccct gtggtcaagc tcgtgggcag gcgcagcctg
     3601 tccatcagct tgtccagcag ggttgtccac gggccgagcg aagcgagcca gccggtggcc
     3661 gctcgcggcc atcgtccaca tatccacggg ctggcaaggg agcgcagcga ccgcgcaggg
     3721 cgaagcccgg agagcaagcc cgtagggggg gcgcgcccag ctgtctaggg cggcggattt
     3781 gtcctactca ggagagcgtt caccgacaaa caacagataa aacgaaaggc ccagtctttc
     3841 gactgagcct ttcgttttat ttgatgcctt taa
//
