LOCUS       sequence-344531-        5783 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CAPTURE method receiver plasmid for Streptomyces heterologous 
            expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5783)
  AUTHORS   Enghiad B, Huang C, Guo F, Jiang G, Wang B, Tabatabaei SK, Martin 
            TA, Zhao H
  TITLE     Cas12a-assisted precise targeted cloning using in vivo Cre-lox 
            recombination.
  JOURNAL   Nat Commun. 2021 Feb 19;12(1):1171. doi: 10.1038/s41467-021-21275-4.
  PUBMED    33608525
REFERENCE   2  (bases 1 to 5783)
  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: "Nat 
            Commun."; date: "2021-02-19"; volume: "12(1):1171. doi"; pages: " 
            10.1038/s41467-021-21275-4"
COMMENT     Sequence Label: pBE45
FEATURES             Location/Qualifiers
     source          1..5783
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(30..47)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(201..789)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(281..300)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(790..823)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             1027..1827
                     /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(2261..2632)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            2665..2774
                     /note="incP origin of transfer"
     CDS             complement(3012..3023)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    3094..3193
                     /label=phage phi-C31 attP
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attachment site of phage phi-C31"
     CDS             3203..5020
                     /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"
     protein_bind    complement(5141..5174)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(5243..5581)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MITNSISRAAADPLESTCSPSLALAVVLQRRDWENPGVTQLNRLA
                     AHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRINSPMSSTSGIG
                     SAADAKCR"
     primer_bind     5482..5504
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5496..5513
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5497..5513
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5582..5598)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5582..5598)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5595..5617)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5606..5622
                     /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(5630..5636,5637..5654,5655..5660))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5675..5696
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
       61 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
      121 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
      181 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
      241 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
      301 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
      361 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
      421 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
      481 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
      541 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
      601 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
      661 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
      721 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
      781 ggatctcaaa taacttcgta tagcatacat tatacgaagt tatgaagatc ctttgatctt
      841 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
      901 attatcaaaa aggatcttca cctagatcct tttggttcat gtgcagctcc atcagcaaaa
      961 ggggatgata agtttatcac caccgactat ttgcaacagt gccgttgatc gtgctatgat
     1021 cgactgatgt catcagcggt ggagtgcaat gtcgtgcaat acgaatggcg aaaagccgag
     1081 ctcatcggtc agcttctcaa ccttggggtt acccccggcg gtgtgctgct ggtccacagc
     1141 tccttccgta gcgtccggcc cctcgaagat gggccacttg gactgatcga ggccctgcgt
     1201 gctgcgctgg gtccgggagg gacgctcgtc atgccctcgt ggtcaggtct ggacgacgag
     1261 ccgttcgatc ctgccacgtc gcccgttaca ccggaccttg gagttgtctc tgacacattc
     1321 tggcgcctgc caaatgtaaa gcgcagcgcc catccatttg cctttgcggc agcggggcca
     1381 caggcagagc agatcatctc tgatccattg cccctgccac ctcactcgcc tgcaagcccg
     1441 gtcgcccgtg tccatgaact cgatgggcag gtacttctcc tcggcgtggg acacgatgcc
     1501 aacacgacgc tgcatcttgc cgagttgatg gcaaaggttc cctatggggt gccgagacac
     1561 tgcaccattc ttcaggatgg caagttggta cgcgtcgatt atctcgagaa tgaccactgc
     1621 tgtgagcgct ttgccttggc ggacaggtgg ctcaaggaga agagccttca gaaggaaggt
     1681 ccagtcggtc atgcctttgc tcggttgatc cgctcccgcg acattgtggc gacagccctg
     1741 ggtcaactgg gccgagatcc gttgatcttc ctgcatccgc cagaggcggg atgcgaagaa
     1801 tgcgatgccg ctcgccagtc gattggctga gctcatgagc ggagaacgag atgacgttgg
     1861 aggggcaagg tcgcgctgat tgctggggca acacgtggag cggatcgggg attgtctttc
     1921 ttcagctcgc tgatgatatg ctgacgctca atgccgtttg gcctccgact aacgaaaatc
     1981 ccgcatttgg acggctgatc cgattggcac ggcggacggc gaatggcgga gcagacgctc
     2041 gtccgggggc aatgagatat gaaaaagcct gaactcaccg cgacgtatcg ggccctggcc
     2101 agctagctag agtcgacctg caggtccccg gggatcggtc ttgccttgct cgtcggtgat
     2161 gtacttcacc agctccgcga agtcgctctt cttgatggag cgcatgggga cgtgcttggc
     2221 aatcacgcgc accccccggc cgttttagcg gctaaaaaag tcatggctct gccctcgggc
     2281 ggaccacgcc catcatgacc ttgccaagct cgtcctgctt ctcttcgatc ttcgccagca
     2341 gggcgaggat cgtggcatca ccgaaccgcg ccgtgcgcgg gtcgtcggtg agccagagtt
     2401 tcagcaggcc gcccaggcgg cccaggtcgc cattgatgcg ggccagctcg cggacgtgct
     2461 catagtccac gacgcccgtg attttgtagc cctggccgac ggccagcagg taggccgaca
     2521 ggctcatgcc ggccgccgcc gccttttcct caatcgctct tcgttcgtct ggaaggcagt
     2581 acaccttgat aggtgggctg cccttcctgg ttggcttggt ttcatcagcc atccgcttgc
     2641 cctcatctgt tacgccggcg gtagccggcc agcctcgcag agcaggattc ccgttgagca
     2701 ccgccaggtg cgaataaggg acagtgaaga aggaacaccc gctcgcgggt gggcctactt
     2761 cacctatcct gcccggctga cgccgttgga tacaccaagg aaagtctaca cgaacccttt
     2821 ggcaaaatcc tgtatatcgt gcgaaaaagg atggatatac cgaaaaaatc gctataatga
     2881 ccccgaagca gggttatgca gcggaaaaga tccgtcgacc tgcaggcatg caagctctag
     2941 cgattccaga cgtcccgaag gcgtggcgcg gcttccccgt gccggagcaa tcgccctggg
     3001 tgggttacac gacgcccctc tatggcccgt actgacggac acaccgaagc cccggcggca
     3061 accctcagcg gatgccccgg ggcttcacgt tttcccaggt cagaagcggt tttcgggagt
     3121 agtgccccaa ctggggtaac ctttgagttc tctcagttgg gggcgtaggg tcgccgacat
     3181 gacacaaggg gttgtgaccg gggtggacac gtacgcgggt gcttacgacc gtcagtcgcg
     3241 cgagcgcgaa aattcgagcg cagcaagccc agcgacacag cgtagcgcca acgaagacaa
     3301 ggcggccgac cttcagcgcg aagtcgagcg cgacgggggc cggttcaggt tcgtcgggca
     3361 tttcagcgaa gcgccgggca cgtcggcgtt cgggacggcg gagcgcccgg agttcgaacg
     3421 catcctgaac gaatgccgcg ccgggcggct caacatgatc attgtctatg acgtgtcgcg
     3481 cttctcgcgc ctgaaggtca tggacgcgat tccgattgtc tcggaattgc tcgccctggg
     3541 cgtgacgatt gtttccactc aggaaggcgt cttccggcag ggaaacgtca tggacctgat
     3601 tcacctgatt atgcggctcg acgcgtcgca caaagaatct tcgctgaagt cggcgaagat
     3661 tctcgacacg aagaaccttc agcgcgaatt gggcgggtac gtcggcggga aggcgcctta
     3721 cggcttcgag cttgtttcgg agacgaagga gatcacgcgc aacggccgaa tggtcaatgt
     3781 cgtcatcaac aagcttgcgc actcgaccac tccccttacc ggacccttcg agttcgagcc
     3841 cgacgtaatc cggtggtggt ggcgtgagat caagacgcac aaacaccttc ccttcaagcc
     3901 gggcagtcaa gccgccattc acccgggcag catcacgggg ctttgtaagc gcatggacgc
     3961 tgacgccgtg ccgacccggg gcgagacgat tgggaagaag accgcttcaa gcgcctggga
     4021 cccggcaacc gttatgcgaa tccttcggga cccgcgtatt gcgggcttcg ccgctgaggt
     4081 gatctacaag aagaagccgg acggcacgcc gaccacgaag attgagggtt accgcattca
     4141 gcgcgacccg atcacgctcc ggccggtcga gcttgattgc ggaccgatca tcgagcccgc
     4201 tgagtggtat gagcttcagg cgtggttgga cggcaggggg cgcggcaagg ggctttcccg
     4261 ggggcaagcc attctgtccg ccatggacaa gctgtactgc gagtgtggcg ccgtcatgac
     4321 ttcgaagcgc ggggaagaat cgatcaagga ctcttaccgc tgccgtcgcc ggaaggtggt
     4381 cgacccgtcc gcacctgggc agcacgaagg cacgtgcaac gtcagcatgg cggcactcga
     4441 caagttcgtt gcggaacgca tcttcaacaa gatcaggcac gccgaaggcg acgaagagac
     4501 gttggcgctt ctgtgggaag ccgcccgacg cttcggcaag ctcactgagg cgcctgagaa
     4561 gagcggcgaa cgggcgaacc ttgttgcgga gcgcgccgac gccctgaacg cccttgaaga
     4621 gctgtacgaa gaccgcgcgg caggcgcgta cgacggaccc gttggcagga agcacttccg
     4681 gaagcaacag gcagcgctga cgctccggca gcaaggggcg gaagagcggc ttgccgaact
     4741 tgaagccgcc gaagccccga agcttcccct tgaccaatgg ttccccgaag acgccgacgc
     4801 tgacccgacc ggccctaagt cgtggtgggg gcgcgcgtca gtagacgaca agcgcgtgtt
     4861 cgtcgggctc ttcgtagaca agatcgttgt cacgaagtcg actacgggca gggggcaggg
     4921 aacgcccatc gagaagcgcg cttcgatcac gtgggcgaag ccgccgaccg acgacgacga
     4981 agacgacgcc caggacggca cggaagacgt agcggcgtag cgagacaccc gggaagcctg
     5041 atctacgtct gtcgagaagt ttctgatcga aaagttcgac agcgtctccg acctgatgca
     5101 gctctcggag ggcgaagaat ctcgtgcttt cagcttcgat ataacttcgt atagcataca
     5161 ttatacgaag ttatgtagga gggcgtggat atgtcctgcg ggtaaatagc tgcgccgatg
     5221 gtttctacaa agatcgttat gtttatcggc actttgcatc ggccgcgctc ccgattccgg
     5281 aagtgcttga cattggggaa tttatgcggt gtgaaatacc gcacagatgc gtaaggagaa
     5341 aataccgcat caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg
     5401 tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa
     5461 gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgccaagc
     5521 ttgggctgca ggtcgactct agaggatccg cggccgcgcg cgatatcgaa ttcgtaatca
     5581 tgtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     5641 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     5701 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     5761 tcggccaacg cgcggggaga ggc
//
