LOCUS       sequence-208047-        2380 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BTK Type 3 coding sequence for golden gate assembly.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2380)
  AUTHORS   Leonard SP, Perutka J, Powell JE, Geng P, Richhart DD, Byrom M, Kar 
            S, Davies BW, Ellington AD, Moran NA, Barrick JE
  TITLE     Genetic Engineering of Bee Gut Microbiome Bacteria with a Toolkit 
            for Modular Assembly of Broad-Host-Range Plasmids.
  JOURNAL   ACS Synth Biol. 2018 May 18;7(5):1279-1290. doi: 
            10.1021/acssynbio.7b00399. Epub 2018 Apr 13.
  PUBMED    29608282
REFERENCE   2  (bases 1 to 2380)
  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.7b00399"; journalName: "ACS Synth Biol"; date: 
            "2018-05-18- 18"; volume: "7"; issue: "5"; pages: "1279-1290"
COMMENT     Sequence Label: pBTK224
FEATURES             Location/Qualifiers
     source          1..2380
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      77..665
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     566..585
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             777..1451
                     /codon_start=1
                     /product="far-red noncytotoxic tetrameric variant of DsRed 
                     fluorescent protein (Strack et al., 2009)"
                     /label=E2-Crimson
                     /note="mammalian codon-optimized"
                     /translation="MDSTENVIKPFMRFKVHMEGSVNGHEFEIEGVGEGKPYEGTQTAK
                     LQVTKGGPLPFAWDILSPQFFYGSKAYIKHPADIPDYLKQSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGTLIYHVKFIGVNFPSDGPVMQKKTLGWEPSTERNYPRDGVLKGENHM
                     ALKLKGGGHYLCEFKSIYMAKKPVKLPGYHYVDYKLDITSHNEDYTVVEQYERAEARHH
                     LFQ"
     primer_bind     1175..1194
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     terminator      complement(1511..1545)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(1616..2275)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2190..2209
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
       61 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
      121 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
      181 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
      241 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
      301 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
      361 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
      421 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
      481 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
      541 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
      601 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
      661 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
      721 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtagtcgg tctcatatgg
      781 atagcactga gaacgtcatc aagcccttca tgcgcttcaa ggtgcacatg gagggctccg
      841 tgaacggcca cgagttcgag atcgagggcg tgggcgaggg caagccctac gagggcaccc
      901 agaccgccaa gctgcaagtg accaagggcg gccccctgcc cttcgcctgg gacatcctgt
      961 ccccccagtt cttctacggc tccaaggcgt acatcaagca ccccgccgac atccccgact
     1021 acctcaagca gtccttcccc gagggcttca agtgggagcg cgtgatgaac ttcgaggacg
     1081 gcggcgtggt gaccgtgacc caggactcct ccctgcagga cggcaccctc atctaccacg
     1141 tgaagttcat cggcgtgaac ttcccctccg acggccccgt aatgcagaag aagactctgg
     1201 gctgggagcc ctccactgag cgcaactacc cccgcgacgg cgtgctgaag ggcgagaacc
     1261 acatggcgct gaagctgaag ggcggcggcc actacctgtg tgagttcaag tccatctaca
     1321 tggccaagaa gcccgtgaag ctgcccggct accactacgt ggactacaag ctcgacatca
     1381 cctcccacaa cgaggactac accgtggtgg agcagtacga gcgcgccgag gcccgccacc
     1441 acctgttcca gactcacggt atggacgaat tgtacaagca cgacgaattg taaatcctga
     1501 gaccagacca ataaaaaacg cccggcggca accgagcgtt ctgaacaaat ccagatggag
     1561 ttctgaggtc attactggat ctatcaacag gagtccaagc gagctcgata tcaaattacg
     1621 ccccgccctg ccactcatcg cagtactgtt gtaattcatt aagcattctg ccgacatgga
     1681 agccatcaca aacggcatga tgaacctgaa tcgccagcgg catcagcacc ttgtcgcctt
     1741 gcgtataata tttgcccatg gtgaaaacgg gggcgaagaa gttgtccata ttggccacgt
     1801 ttaaatcaaa actggtgaaa ctcacccagg gattggctga aacgaaaaac atattctcaa
     1861 taaacccttt agggaaatag gccaggtttt caccgtaaca cgccacatct tgcgaatata
     1921 tgtgtagaaa ctgccggaaa tcgtcgtggt attcactcca gagcgatgaa aacgtttcag
     1981 tttgctcatg gaaaacggtg taacaagggt gaacactatc ccatatcacc agctcaccgt
     2041 ctttcattgc catacgaaat tccggatgag cattcatcag gcgggcaaga atgtgaataa
     2101 aggccggata aaacttgtgc ttatttttct ttacggtctt taaaaaggcc gtaatatcca
     2161 gctgaacggt ctggttatag gtacattgag caactgactg aaatgcctca aaatgttctt
     2221 tacgatgcca ttgggatata tcaacggtgg tatatccagt gatttttttc tccattttag
     2281 cttccttagc tcctgaaaat ctcgataact caaaaaatac gcccggtagt gatcttattt
     2341 cattatggtg aaagttggaa cctcttacgt gcccgatcaa
//
