LOCUS       sequence-305129-        6705 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 6705)
  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 6705)
  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: pSETT4tip
FEATURES             Location/Qualifiers
     source          1..6705
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(595..1183)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(675..694)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1387..2187
                     /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(2621..2992)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            3025..3134
                     /note="incP origin of transfer"
     CDS             complement(3372..3383)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    3454..3553
                     /label=phage phi-C31 attP
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attachment site of phage phi-C31"
     CDS             3563..5380
                     /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(5799..5816)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     CDS             complement(6066..6410)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MITNSISRAAADPLESTCSPSLALAVVLQRRDWENPGVTQLNRLA
                     AHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRINSPMSSTSGIG
                     SAADAKCRST"
     primer_bind     6311..6333
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6325..6342
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6326..6342
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6411..6427)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6411..6427)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6424..6446)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6435..6451
                     /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(6459..6465,6466..6483,6484..6489))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6504..6525
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 cctctagggt ccccaattaa ttagtaatat aatctattaa aggtcattca aaaggtcatc
       61 cactgcaggt gtaagtgtgc caagtctaag gactacgatg ggttttgttg gaccttacag
      121 atctggagtc ggtctaattt aaatggtgct gagcatttgt gcgccaaaag accgtcacat
      181 agagcgcata cccggaagcg acgaatgtct caggtatcca aaaagtaacc ctgatgccgc
      241 gcttcgattc accgaagtga ggtggatgac cttttgaatg acctttaata gattatatta
      301 ctaattaatt ggggacccta gaggtcccct tttttatttt aaaaattttt tcacaaaacg
      361 gtttacaagc ataaagcttg ccacgcagac gacagcccac gctgaccgat ctacctgaac
      421 ggcgaccatc tgtgtggtac tggggcggag agataactac ggtgccgctt accgggctca
      481 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
      541 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
      601 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
      661 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
      721 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
      781 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
      841 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
      901 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
      961 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     1021 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     1081 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     1141 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     1201 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     1261 attatcaaaa aggatcttca cctagatcct tttggttcat gtgcagctcc atcagcaaaa
     1321 ggggatgata agtttatcac caccgactat ttgcaacagt gccgttgatc gtgctatgat
     1381 cgactgatgt catcagcggt ggagtgcaat gtcgtgcaat acgaatggcg aaaagccgag
     1441 ctcatcggtc agcttctcaa ccttggggtt acccccggcg gtgtgctgct ggtccacagc
     1501 tccttccgta gcgtccggcc cctcgaagat gggccacttg gactgatcga ggccctgcgt
     1561 gctgcgctgg gtccgggagg gacgctcgtc atgccctcgt ggtcaggtct ggacgacgag
     1621 ccgttcgatc ctgccacgtc gcccgttaca ccggaccttg gagttgtctc tgacacattc
     1681 tggcgcctgc caaatgtaaa gcgcagcgcc catccatttg cctttgcggc agcggggcca
     1741 caggcagagc agatcatctc tgatccattg cccctgccac ctcactcgcc tgcaagcccg
     1801 gtcgcccgtg tccatgaact cgatgggcag gtacttctcc tcggcgtggg acacgatgcc
     1861 aacacgacgc tgcatcttgc cgagttgatg gcaaaggttc cctatggggt gccgagacac
     1921 tgcaccattc ttcaggatgg caagttggta cgcgtcgatt atctcgagaa tgaccactgc
     1981 tgtgagcgct ttgccttggc ggacaggtgg ctcaaggaga agagccttca gaaggaaggt
     2041 ccagtcggtc atgcctttgc tcggttgatc cgctcccgcg acattgtggc gacagccctg
     2101 ggtcaactgg gccgagatcc gttgatcttc ctgcatccgc cagaggcggg atgcgaagaa
     2161 tgcgatgccg ctcgccagtc gattggctga gctcatgagc ggagaacgag atgacgttgg
     2221 aggggcaagg tcgcgctgat tgctggggca acacgtggag cggatcgggg attgtctttc
     2281 ttcagctcgc tgatgatatg ctgacgctca atgtcgtttg gcctccgact aacgaaaatc
     2341 ccgcatttgg acggctgatc cgattggcac ggcggacggc gaatggcgga gcagacgctc
     2401 gtccgggggc aatgagatat gaaaaagcct gaactcaccg cgacgtatcg ggccctggcc
     2461 agctagctag agtcgacctg caggtccccg gggatcggtc ttgccttgct cgtcggtgat
     2521 gtacttcacc agctccgcga agtcgctctt cttgatggag cgcatgggga cgtgcttggc
     2581 aatcacgcgc accccccggc cgttttagcg gctaaaaaag tcatggctct gccctcgggc
     2641 ggaccacgcc catcatgacc ttgccaagct cgtcctgctt ctcttcgatc ttcgccagca
     2701 gggcgaggat cgtggcatca ccgaaccgcg ccgtgcgcgg gtcgtcggtg agccagagtt
     2761 tcagcaggcc gcccaggcgg cccaggtcgc cattgatgcg ggccagctcg cggacgtgct
     2821 catagtccac gacgcccgtg attttgtagc cctggccgac ggccagcagg taggccgaca
     2881 ggctcatgcc ggccgccgcc gccttttcct caatcgctct tcgttcgtct ggaaggcagt
     2941 acaccttgat aggtgggctg cccttcctgg ttggcttggt ttcatcagcc atccgcttgc
     3001 cctcatctgt tacgccggcg gtagccggcc agcctcgcag agcaggattc ccgttgagca
     3061 ccgccaggtg cgaataaggg acagtgaaga aggaacaccc gctcgcgggt gggcctactt
     3121 cacctatcct gcccggctga cgccgttgga tacaccaagg aaagtctaca cgaacccttt
     3181 ggcaaaatcc tgtatatcgt gcgaaaaagg atggatatac cgaaaaaatc gctataatga
     3241 ccccgaagca gggttatgca gcggaaaaga tccgtcgacc tgcaggcatg caagctctag
     3301 cgattccaga cgtcccgaag gcgtggcgcg gcttccccgt gccggagcaa tcgccctggg
     3361 tgggttacac gacgcccctc tatggcccgt actgacggac acaccgaagc cccggcggca
     3421 accctcagcg gatgccccgg ggcttcacgt tttcccaggt cagaagcggt tttcgggagt
     3481 agtgccccaa ctggggtaac ctttgagttc tctcagttgg gggcgtaggg tcgccgacat
     3541 gacacaaggg gttgtgaccg gggtggacac gtacgcgggt gcttacgacc gtcagtcgcg
     3601 cgagcgcgaa aattcgagcg cagcaagccc agcgacacag cgtagcgcca acgaagacaa
     3661 ggcggccgac cttcagcgcg aagtcgagcg cgacgggggc cggttcaggt tcgtcgggca
     3721 tttcagcgaa gcgccgggca cgtcggcgtt cgggacggcg gagcgcccgg agttcgaacg
     3781 catcctgaac gaatgccgcg ccgggcggct caacatgatc attgtctatg acgtgtcgcg
     3841 cttctcgcgc ctgaaggtca tggacgcgat tccgattgtc tcggaattgc tcgccctggg
     3901 cgtgacgatt gtttccactc aggaaggcgt cttccggcag ggaaacgtca tggacctgat
     3961 tcacctgatt atgcggctcg acgcgtcgca caaagaatct tcgctgaagt cggcgaagat
     4021 tctcgacacg aagaaccttc agcgcgaatt gggcgggtac gtcggcggga aggcgcctta
     4081 cggcttcgag cttgtttcgg agacgaagga gatcacgcgc aacggccgaa tggtcaatgt
     4141 cgtcatcaac aagcttgcgc actcgaccac tccccttacc ggacccttcg agttcgagcc
     4201 cgacgtaatc cggtggtggt ggcgtgagat caagacgcac aaacaccttc ccttcaagcc
     4261 gggcagtcaa gccgccattc acccgggcag catcacgggg ctttgtaagc gcatggacgc
     4321 tgacgccgtg ccgacccggg gcgagacgat tgggaagaag accgcttcaa gcgcctggga
     4381 cccggcaacc gttatgcgaa tccttcggga cccgcgtatt gcgggcttcg ccgctgaggt
     4441 gatctacaag aagaagccgg acggcacgcc gaccacgaag attgagggtt accgcattca
     4501 gcgcgacccg atcacgctcc ggccggtcga gcttgattgc ggaccgatca tcgagcccgc
     4561 tgagtggtat gagcttcagg cgtggttgga cggcaggggg cgcggcaagg ggctttcccg
     4621 ggggcaagcc attctgtccg ccatggacaa gctgtactgc gagtgtggcg ccgtcatgac
     4681 ttcgaagcgc ggggaagaat cgatcaagga ctcttaccgc tgccgtcgcc ggaaggtggt
     4741 cgacccgtcc gcacctgggc agcacgaagg cacgtgcaac gtcagcatgg cggcactcga
     4801 caagttcgtt gcggaacgca tcttcaacaa gatcaggcac gccgaaggcg acgaagagac
     4861 gttggcgctt ctgtgggaag ccgcccgacg cttcggcaag ctcactgagg cgcctgagaa
     4921 gagcggcgaa cgggcgaacc ttgttgcgga gcgcgccgac gccctgaacg cccttgaaga
     4981 gctgtacgaa gaccgcgcgg caggcgcgta cgacggaccc gttggcagga agcacttccg
     5041 gaagcaacag gcagcgctga cgctccggca gcaaggggcg gaagagcggc ttgccgaact
     5101 tgaagccgcc gaagccccga agcttcccct tgaccaatgg ttccccgaag acgccgacgc
     5161 tgacccgacc ggccctaagt cgtggtgggg gcgcgcgtca gtagacgaca agcgcgtgtt
     5221 cgtcgggctc ttcgtagaca agatcgttgt cacgaagtcg actacgggca gggggcaggg
     5281 aacgcccatc gagaagcgcg cttcgatcac gtgggcgaag ccgccgaccg acgacgacga
     5341 agacgacgcc caggacggca cggaagacgt agcggcgtag cgagacaccc gggaagcctg
     5401 atctacgtct gtcgagaagt ttctgatcga aaagttcgac agcgtctccg acctgatgca
     5461 gctctcggag ggcgaagaat ctcgtgcttt cagcttcgat gtaggagggc gtggatatgt
     5521 cctgcgggta aatagctgcg ccgatggttc tctgtcgtcg ctgacgtctg tagtctagcc
     5581 tcattatgat tgtacgctat tcagggattg actgataccg gaagacatct caaatgaagt
     5641 ggtcaagctt tatgcttgta aaccgttttg tgaaaaaatt tttaaaataa aaaaggggac
     5701 ctctagggtc cccaattaat tagtaatata atctattaaa ggtcattcaa aaggtcatcc
     5761 aagcttggct gttttggcgg atgagagaag attttcagcc tgatacagat taaatcagaa
     5821 cgcagaagcg gtctgataaa acagaatttg cctggcggca gtagcgcggt ggtcccacct
     5881 gaccccatgc cgaactcaga agtgaaacgc cgtagcgccg atggtagtgt ggcccatgcg
     5941 agagtacaat ccctagaacg tccgggcttg cacctcacgt cacgtgagga ggcagcgtgg
     6001 acggcgtggt accaagctta ttggcactag tcgagcaacg gaggtattcc gatggtaccg
     6061 agaccttatg ttgatcggca ctttgcatcg gccgcgctcc cgattccgga agtgcttgac
     6121 attggggaat ttatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
     6181 aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
     6241 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg
     6301 ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgccaagct tgggctgcag
     6361 gtcgactcta gaggatccgc ggccgcgcgc gatatcgaat tcgtaatcat gtcatagctg
     6421 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     6481 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgcggt
     6541 ctcggcgttt cgtgccgcgt gattttccgc caaaaacttt aacgaacgtt cgttataatg
     6601 gtgtcatgac cttcacgacg aagtactaaa attggcccga atcatcagct aagctttatg
     6661 cttgtaaacc gttttgtgaa aaaattttta aaataaaaaa gggga
//
