LOCUS       sequence-305128-        6815 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  phi-C31-integrated plasmid including BsaI-recognition sites and 
            T4-terminators surrounding the expression cassette.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6815)
  AUTHORS   Schaffert L, Jacob L, Schneiker-Bekel S, Persicke M, Marz C, Ruckert
            C, Puhler A, Kalinowski J
  TITLE     pSETT4, an Improved phiC31-Based Integrative Vector System for 
            Actinoplanes sp. SE50/110.
  JOURNAL   Microbiol Resour Announc. 2020 Sep 24;9(39). pii: 9/39/e00596-20. 
            doi: 10.1128/MRA.00596-20.
  PUBMED    32972929
REFERENCE   2  (bases 1 to 6815)
  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 
            Resour Announc."; date: "2020-09-24"; volume: "9(39). pii: 
            9/39/e00596-20. doi"; pages: " 10.1128/MRA.00596-20"
COMMENT     Sequence Label: pSETT4gap
FEATURES             Location/Qualifiers
     source          1..6815
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(581..1169)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(661..680)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1373..2173
                     /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"
     CDS             complement(2607..2978)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            3011..3120
                     /note="incP origin of transfer"
     CDS             complement(3358..3369)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    3440..3539
                     /label=phage phi-C31 attP
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attachment site of phage phi-C31"
     CDS             3549..5366
                     /codon_start=1
                     /product="integrase from phage phi-C31"
                     /label=phage phi-C31 integrase
                     /translation="MDTYAGAYDRQSRERENSSAASPATQRSANEDKAADLQREVERDG
                     GRFRFVGHFSEAPGTSAFGTAERPEFERILNECRAGRLNMIIVYDVSRFSRLKVMDAIP
                     IVSELLALGVTIVSTQEGVFRQGNVMDLIHLIMRLDASHKESSLKSAKILDTKNLQREL
                     GGYVGGKAPYGFELVSETKEITRNGRMVNVVINKLAHSTTPLTGPFEFEPDVIRWWWRE
                     IKTHKHLPFKPGSQAAIHPGSITGLCKRMDADAVPTRGETIGKKTASSAWDPATVMRIL
                     RDPRIAGFAAEVIYKKKPDGTPTTKIEGYRIQRDPITLRPVELDCGPIIEPAEWYELQA
                     WLDGRGRGKGLSRGQAILSAMDKLYCECGAVMTSKRGEESIKDSYRCRRRKVVDPSAPG
                     QHEGTCNVSMAALDKFVAERIFNKIRHAEGDEETLALLWEAARRFGKLTEAPEKSGERA
                     NLVAERADALNALEELYEDRAAGAYDGPVGRKHFRKQQAALTLRQQGAEERLAELEAAE
                     APKLPLDQWFPEDADADPTGPKSWWGRASVDDKRVFVGLFVDKIVVTKSTTGRGQGTPI
                     EKRASITWAKPPTDDDEDDAQDGTEDVAA"
     primer_bind     complement(5785..5802)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     CDS             complement(6162..6506)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MITNSISRAAADPLESTCSPSLALAVVLQRRDWENPGVTQLNRLA
                     AHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRINSPMSSTSGIG
                     SAANAKCRST"
     primer_bind     6407..6429
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6421..6438
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6422..6438
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6507..6523)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6507..6523)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6520..6542)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6531..6547
                     /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(6555..6561,6562..6579,6580..6585))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6600..6621
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 aattaattag taatataatc tattaaaggt cattcaaaag gtcatccacc tcacttcggt
       61 gaatcgaagc gcggcatcag ggttactttt tggatacctg agacattcgt cgcttccggg
      121 tatgcgctct atgtgacggt cttttggcgc acaaatgctc agcaccattt aaattagacc
      181 gactccagat ctgtaaggtc caacaaaacc catcgtagtc cttagacttg gcacacttac
      241 acctgcagtg gatgaccttt tgaatgacct ttaatagatt atattactaa ttaattgggg
      301 accctagagg tccccttttt tattttaaaa attttttcac aaaacggttt acaagcataa
      361 agcttgccac gcagacgaca gcccacgctg accgatctac ctgaacggcg accatctgtg
      421 tggtactggg gcggagagat aactacggtg ccgcttaccg ggctcactca aaggcggtaa
      481 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
      541 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
      601 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
      661 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
      721 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
      781 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
      841 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
      901 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
      961 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     1021 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     1081 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     1141 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     1201 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     1261 tcttcaccta gatccttttg gttcatgtgc agctccatca gcaaaagggg atgataagtt
     1321 tatcaccacc gactatttgc aacagtgccg ttgatcgtgc tatgatcgac tgatgtcatc
     1381 agcggtggag tgcaatgtcg tgcaatacga atggcgaaaa gccgagctca tcggtcagct
     1441 tctcaacctt ggggttaccc ccggcggtgt gctgctggtc cacagctcct tccgtagcgt
     1501 ccggcccctc gaagatgggc cacttggact gatcgaggcc ctgcgtgctg cgctgggtcc
     1561 gggagggacg ctcgtcatgc cctcgtggtc aggtctggac gacgagccgt tcgatcctgc
     1621 cacgtcgccc gttacaccgg accttggagt tgtctctgac acattctggc gcctgccaaa
     1681 tgtaaagcgc agcgcccatc catttgcctt tgcggcagcg gggccacagg cagagcagat
     1741 catctctgat ccattgcccc tgccacctca ctcgcctgca agcccggtcg cccgtgtcca
     1801 tgaactcgat gggcaggtac ttctcctcgg cgtgggacac gatgccaaca cgacgctgca
     1861 tcttgccgag ttgatggcaa aggttcccta tggggtgccg agacactgca ccattcttca
     1921 ggatggcaag ttggtacgcg tcgattatct cgagaatgac cactgctgtg agcgctttgc
     1981 cttggcggac aggtggctca aggagaagag ccttcagaag gaaggtccag tcggtcatgc
     2041 ctttgctcgg ttgatccgct cccgcgacat tgtggcgaca gccctgggtc aactgggccg
     2101 agatccgttg atcttcctgc atccgccaga ggcgggatgc gaagaatgcg atgccgctcg
     2161 ccagtcgatt ggctgagctc atgagcggag aacgagatga cgttggaggg gcaaggtcgc
     2221 gctgattgct ggggcaacac gtggagcgga tcggggattg tctttcttca gctcgctgat
     2281 gatatgctga cgctcaatgc cgtttggcct ccgactaacg aaaatcccgc atttggacgg
     2341 ctgatccgat tggcacggcg gacggcgaat ggcggagcag acgctcgtcc gggggcaatg
     2401 agatatgaaa aagcctgaac tcaccgcgac gtatcgggcc ctggccagct agctagagtc
     2461 gacctgcagg tccccgggga tcggtcttgc cttgctcgtc ggtgatgtac ttcaccagct
     2521 ccgcgaagtc gctcttcttg atggagcgca tggggacgtg cttggcaatc acgcgcaccc
     2581 cccggccgtt ttagcggcta aaaaagtcat ggctctgccc tcgggcggac cacgcccatc
     2641 atgaccttgc caagctcgtc ctgcttctct tcgatcttcg ccagcagggc gaggatcgtg
     2701 gcatcaccga accgcgccgt gcgcgggtcg tcggtgagcc agagtttcag caggccgccc
     2761 aggcggccca ggtcgccatt gatgcgggcc agctcgcgga cgtgctcata gtccacgacg
     2821 cccgtgattt tgtagccctg gccgacggcc agcaggtagg ccgacaggct catgccggcc
     2881 gccgccgcct tttcctcaat cgctcttcgt tcgtctggaa ggcagtacac cttgataggt
     2941 gggctgccct tcctggttgg cttggtttca tcagccatcc gcttgccctc atctgttacg
     3001 ccggcggtag ccggccagcc tcgcagagca ggattcccgt tgagcaccgc caggtgcgaa
     3061 taagggacag tgaagaagga acacccgctc gcgggtgggc ctacttcacc tatcctgccc
     3121 ggctgacgcc gttggataca ccaaggaaag tctacacgaa ccctttggca aaatcctgta
     3181 tatcgtgcga aaaaggatgg atataccgaa aaaatcgcta taatgacccc gaagcagggt
     3241 tatgcagcgg aaaagatccg tcgacctgca ggcatgcaag ctctagcgat tccagacgtc
     3301 ccgaaggcgt ggcgcggctt ccccgtgccg gagcaatcgc cctgggtggg ttacacgacg
     3361 cccctctatg gcccgtactg acggacacac cgaagccccg gcggcaaccc tcagcggatg
     3421 ccccggggct tcacgttttc ccaggtcaga agcggttttc gggagtagtg ccccaactgg
     3481 ggtaaccttt gagttctctc agttgggggc gtagggtcgc cgacatgaca caaggggttg
     3541 tgaccggggt ggacacgtac gcgggtgctt acgaccgtca gtcgcgcgag cgcgaaaatt
     3601 cgagcgcagc aagcccagcg acacagcgta gcgccaacga agacaaggcg gccgaccttc
     3661 agcgcgaagt cgagcgcgac gggggccggt tcaggttcgt cgggcatttc agcgaagcgc
     3721 cgggcacgtc ggcgttcggg acggcggagc gcccggagtt cgaacgcatc ctgaacgaat
     3781 gccgcgccgg gcggctcaac atgatcattg tctatgacgt gtcgcgcttc tcgcgcctga
     3841 aggtcatgga cgcgattccg attgtctcgg aattgctcgc cctgggcgtg acgattgttt
     3901 ccactcagga aggcgtcttc cggcagggaa acgtcatgga cctgattcac ctgattatgc
     3961 ggctcgacgc gtcgcacaaa gaatcttcgc tgaagtcggc gaagattctc gacacgaaga
     4021 accttcagcg cgaattgggc gggtacgtcg gcgggaaggc gccttacggc ttcgagcttg
     4081 tttcggagac gaaggagatc acgcgcaacg gccgaatggt caatgtcgtc atcaacaagc
     4141 ttgcgcactc gaccactccc cttaccggac ccttcgagtt cgagcccgac gtaatccggt
     4201 ggtggtggcg tgagatcaag acgcacaaac accttccctt caagccgggc agtcaagccg
     4261 ccattcaccc gggcagcatc acggggcttt gtaagcgcat ggacgctgac gccgtgccga
     4321 cccggggcga gacgattggg aagaagaccg cttcaagcgc ctgggacccg gcaaccgtta
     4381 tgcgaatcct tcgggacccg cgtattgcgg gcttcgccgc tgaggtgatc tacaagaaga
     4441 agccggacgg cacgccgacc acgaagattg agggttaccg cattcagcgc gacccgatca
     4501 cgctccggcc ggtcgagctt gattgcggac cgatcatcga gcccgctgag tggtatgagc
     4561 ttcaggcgtg gttggacggc agggggcgcg gcaaggggct ttcccggggg caagccattc
     4621 tgtccgccat ggacaagctg tactgcgagt gtggcgccgt catgacttcg aagcgcgggg
     4681 aagaatcgat caaggactct taccgctgcc gtcgccggaa ggtggtcgac ccgtccgcac
     4741 ctgggcagca cgaaggcacg tgcaacgtca gcatggcggc actcgacaag ttcgttgcgg
     4801 aacgcatctt caacaagatc aggcacgccg aaggcgacga agagacgttg gcgcttctgt
     4861 gggaagccgc ccgacgcttc ggcaagctca ctgaggcgcc tgagaagagc ggcgaacggg
     4921 cgaaccttgt tgcggagcgc gccgacgccc tgaacgccct tgaagagctg tacgaagacc
     4981 gcgcggcagg cgcgtacgac ggacccgttg gcaggaagca cttccggaag caacaggcag
     5041 cgctgacgct ccggcagcaa ggggcggaag agcggcttgc cgaacttgaa gccgccgaag
     5101 ccccgaagct tccccttgac caatggttcc ccgaagacgc cgacgctgac ccgaccggcc
     5161 ctaagtcgtg gtgggggcgc gcgtcagtag acgacaagcg cgtgttcgtc gggctcttcg
     5221 tagacaagat cgttgtcacg aagtcgacta cgggcagggg gcagggaacg cccatcgaga
     5281 agcgcgcttc gatcacgtgg gcgaagccgc cgaccgacga cgacgaagac gacgcccagg
     5341 acggcacgga agacgtagcg gcgtagcgag acacccggga agcctgatct acgtctgtcg
     5401 agaagtttct gatcgaaaag ttcgacagcg tctccgacct gatgcagctc tcggagggcg
     5461 aagaatctcg tgctttcagc ttcgatgtag gagggcgtgg atatgtcctg cgggtaaata
     5521 gctgcgccga tggttctctg tcgtcgctga cgtctgtagt ctagcctcat tatgattgta
     5581 cgctattcag ggattgactg ataccggaag acatctcaaa tgaagtggtc aagctttatg
     5641 cttgtaaacc gttttgtgaa aaaattttta aaataaaaaa ggggacctct agggtcccca
     5701 attaattagt aatataatct attaaaggtc attcaaaagg tcatccaagc ttggctgttt
     5761 tggcggatga gagaagattt tcagcctgat acagattaaa tcagaacgca gaagcggtct
     5821 gataaaacag aatttgcctg gcggcagtag cgcggtggtt ccacctgacc ccatgccgaa
     5881 ctcagaagtg aaacgccgta gcgccgatgg tagtgtggcc catgcgagag tacatatggt
     5941 actggccgat gctgggagaa gcgcgctgct gtacggcgcg caccgggtgc ggagcccctc
     6001 ggcgagcggt gtgaaacttc tgtgaatggc ctgttcggtt gcttttttta tacggctgcc
     6061 agataaggct tgcagcatct gggcggctac cgctatgatc ggggcgttcc tgcaattctt
     6121 agtgcgagta tctgaaaggg gatacgcatg gtaccgagac cttatgttga tcggcacttt
     6181 gcattggccg cgctcccgat tccggaagtg cttgacattg gggaatttat gcggtgtgaa
     6241 ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc attcaggctg
     6301 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa
     6361 gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt
     6421 tgtaaaacga cggccagtgc caagcttggg ctgcaggtcg actctagagg atccgcggcc
     6481 gcgcgcgata tcgaattcgt aatcatgtca tagctgtttc ctgtgtgaaa ttgttatccg
     6541 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     6601 tgagtgagct aactcacatt aattgcgttg cgcggtctcg gcgtttcgtg ccgcgtgatt
     6661 ttccgccaaa aactttaacg aacgttcgtt ataatggtgt catgaccttc acgacgaagt
     6721 actaaaattg gcccgaatca tcagctaagc tttatgcttg taaaccgttt tgtgaaaaaa
     6781 tttttaaaat aaaaaagggg acctctaggg tcccc
//
