LOCUS       sequence-338644-        3665 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Suicide vector carrying the site attL of the SSR system of pSAM2. 
            This plasmid is integrated into the genome of Actinobacteria by HR 
            when carrying a DNA fragment identical to a region of the genome..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3665)
  AUTHORS   Santos LDF, Caraty-Philippe L, Darbon E, Pernodet JL
  TITLE     Marker-Free Genome Engineering in Amycolatopsis Using the pSAM2 
            Site-Specific Recombination System.
  JOURNAL   Microorganisms. 2022 Apr 16;10(4). pii: microorganisms10040828. doi:
            10.3390/microorganisms10040828.
  PUBMED    35456877
REFERENCE   2  (bases 1 to 3665)
  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: 
            "Microorganisms."; date: "2022-04-16"; volume: "10(4). pii: 
            microorganisms10040828. doi"; pages: " 
            10.3390/microorganisms10040828"
COMMENT     Sequence Label: pEA02
FEATURES             Location/Qualifiers
     source          1..3665
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     463..485
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     477..494
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     478..494
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(759..775)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(759..775)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(772..794)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    783..799
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(807..813,814..831,832..837))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    852..873
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(990..1007)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1161..1749)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1241..1260)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1953..2753
                     /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            3198..3307
                     /note="incP origin of transfer"
ORIGIN
        1 ggtgaagtac atcaccgacg agcaaggcaa gaccgatccc cggggacctg catagatcta
       61 cgtctgtcga gaagtttctg atcgaaaagt tcgacagcgt ctccgacctg atgcagctct
      121 cggagggcga agaatctcgt gctttcagct tcgatgtagg agggcgtgga tatgtcctgc
      181 gggtaaatag ctgcgccgat ggtttctaca aagatcgtta tgtttatcgg cactttgcat
      241 cggccgcgct cccgattccg gaagtgcttg acattgggga atttatgcgg tgtgaaatac
      301 cgcacagatg cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca
      361 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg
      421 gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta
      481 aaacgacggc cagtgccaag cttgggctgc aggtcgactc tagaggatcc ctgtcagtca
      541 tgcgggcaac ttagccgagc cctacgacac cgcccgtggg aaggtgagtg tctaactgcg
      601 tgacaacgcc agcgcacagc ggcggacaac cgcgagcacc catggactgg cgccgcaggt
      661 gagaagcaca ctggcccaag gtcgagcgcc cacccaagtt gcttcgggac gaagaggtag
      721 atatctatcc taggcggccg catcgaattc gtaatcatgt catagctgtt tcctgtgtga
      781 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
      841 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
      901 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
      961 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     1021 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     1081 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     1141 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     1201 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     1261 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     1321 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     1381 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     1441 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     1501 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     1561 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     1621 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     1681 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     1741 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     1801 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatccttttg
     1861 gttcatgtgc agctccatca gcaaaagggg atgataagtt tatcaccacc gactatttgc
     1921 aacagtgccg ttgatcgtgc tatgatcgac tgatgtcatc agcggtggag tgcaatgtcg
     1981 tgcaatacga atggcgaaaa gccgagctca tcggtcagct tctcaacctt ggggttaccc
     2041 ccggcggtgt gctgctggtc cacagctcct tccgtagcgt ccggcccctc gaagatgggc
     2101 cacttggact gatcgaggcc ctgcgtgctg cgctgggtcc gggagggacg ctcgtcatgc
     2161 cctcgtggtc aggtctggac gacgagccgt tcgatcctgc cacgtcgccc gttacaccgg
     2221 accttggagt tgtctctgac acattctggc gcctgccaaa tgtaaagcgc agcgcccatc
     2281 catttgcctt tgcggcagcg gggccacagg cagagcagat catctctgat ccattgcccc
     2341 tgccacctca ctcgcctgca agcccggtcg cccgtgtcca tgaactcgat gggcaggtac
     2401 ttctcctcgg cgtgggacac gatgccaaca cgacgctgca tcttgccgag ttgatggcaa
     2461 aggttcccta tggggtgccg agacactgca ccattcttca ggatggcaag ttggtacgcg
     2521 tcgattatct cgagaatgac cactgctgtg agcgctttgc cttggcggac aggtggctca
     2581 aggagaagag ccttcagaag gaaggtccag tcggtcatgc ctttgctcgg ttgatccgct
     2641 cccgcgacat tgtggcgaca gccctgggtc aactgggccg agatccgttg atcttcctgc
     2701 atccgccaga ggcgggatgc gaagaatgcg atgccgctcg ccagtcgatt ggctgagctc
     2761 atgagcggag aacgagatga cgttggaggg gcaaggtcgc gctgattgct ggggcaacac
     2821 gtggagcgga tcggggattg tctttcttca gctcgctgat gatatgctga cgctcaatgc
     2881 cgtttggcct ccgactaacg aaaatcccgc atttggacgg ctgatccgat tggcacggcg
     2941 gacggcgaat ggcggagcag acgctcgtcc gggggcaatg agatatgaaa aagcctgaac
     3001 tcaccgcgac gtatcgatgg taccatatat ttaaagatcc actagtatgc aggtcgacgg
     3061 atcttttccg ctgcataacc ctgcttcggg gtcattatag cgattttttc ggtatatcca
     3121 tcctttttcg cacgatatac aggattttgc caaagggttc gtgtagactt tccttggtgt
     3181 atccaacggc gtcagccggg caggataggt gaagtaggcc cacccgcgag cgggtgttcc
     3241 ttcttcactg tcccttattc gcacctggcg gtgctcaacg ggaatcctgc tctgcgaggc
     3301 tggccggcat gagcctgtcg gcctacctgc tggccgtcgg ccagggctac aaaatcacgg
     3361 gcgtcgtgga ctatgagcac gtccgcgagc tggcccgcat caatggcgac ctgggccgcc
     3421 tgggcggcct gctgaaactc tggctcaccg acgacccgcg cacggcgcgg ttcggtgatg
     3481 ccacgatcct cgccctgctg gcgaagatcg aagagaagca ggacgagctt ggcaaggtca
     3541 tgatgggcgt ggtccgcccg agggcagagc catgactttt ttagccgcta aaacggccgg
     3601 ggggtgcgcg tgattgccaa gcacgtcccc atgcgctcca tcaagaagag cgacttcgcg
     3661 gagct
//
