LOCUS       sequence-249005-        1719 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes BxBI attP site as a Type 3a part to be used in the MTK 
            system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 1719)
  AUTHORS   Fonseca JP, Bonny AR, Kumar GR, Ng AH, Town J, Wu QC, Aslankoohi E, 
            Chen SY, Dods G, Harrigan P, Osimiri LC, Kistler AL, El-Samad H
  TITLE     A Toolkit for Rapid Modular Construction of Biological Circuits in 
            Mammalian Cells.
  JOURNAL   ACS Synth Biol. 2019 Nov 15;8(11):2593-2606. doi: 
            10.1021/acssynbio.9b00322. Epub 2019 Nov 5.
  PUBMED    31686495
REFERENCE   2  (bases 1 to 1719)
  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"; doi: 
            "10.1021/acssynbio.9b00322"; journalName: "ACS Synth Biol"; date: 
            "2019-11-15- 15"; volume: "8"; issue: "11"; pages: "2593-2606"
COMMENT     Sequence Label: MTK3a_009
FEATURES             Location/Qualifiers
     source          1..1719
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      56..644
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     545..564
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    761..808
                     /label=Bxb1 attP
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="Minimal attP site for the Mycopbacteriophage Bxb1 
                     integrase (Voutev and Mann, 2017)"
     terminator      complement(829..863)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(934..1593)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1508..1527
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
       61 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
      121 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
      181 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
      241 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
      301 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
      361 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
      421 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
      481 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
      541 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
      601 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
      661 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
      721 cccctgattc tgtggataac cgtagtcggt ctcatatggt ggtttgtctg gtcaaccacc
      781 gcggtctcag tggtgtacgg tacaaaccag gttcttgaga ccagaccaat aaaaaacgcc
      841 cggcggcaac cgagcgttct gaacaaatcc agatggagtt ctgaggtcat tactggatct
      901 atcaacagga gtccaagcga gctcgatatc aaattacgcc ccgccctgcc actcatcgca
      961 gtactgttgt aattcattaa gcattctgcc gacatggaag ccatcacaaa cggcatgatg
     1021 aacctgaatc gccagcggca tcagcacctt gtcgccttgc gtataatatt tgcccatggt
     1081 gaaaacgggg gcgaagaagt tgtccatatt ggccacgttt aaatcaaaac tggtgaaact
     1141 cacccaggga ttggctgaaa cgaaaaacat attctcaata aaccctttag ggaaataggc
     1201 caggttttca ccgtaacacg ccacatcttg cgaatatatg tgtagaaact gccggaaatc
     1261 gtcgtggtat tcactccaga gcgatgaaaa cgtttcagtt tgctcatgga aaacggtgta
     1321 acaagggtga acactatccc atatcaccag ctcaccgtct ttcattgcca tacgaaattc
     1381 cggatgagca ttcatcaggc gggcaagaat gtgaataaag gccggataaa acttgtgctt
     1441 atttttcttt acggtcttta aaaaggccgt aatatccagc tgaacggtct ggttataggt
     1501 acattgagca actgactgaa atgcctcaaa atgttcttta cgatgccatt gggatatatc
     1561 aacggtggta tatccagtga tttttttctc cattttagct tccttagctc ctgaaaatct
     1621 cgataactca aaaaatacgc ccggtagtga tcttatttca ttatggtgaa agttggaacc
     1681 tcttacgtgc ccgatcaatc atgaccaaaa tcccttaac
//
