LOCUS       sequence-328051-        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"
     CDS             99..773
                     /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     497..516
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     terminator      complement(833..867)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(938..1597)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1512..1531
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      1779..2367
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2268..2287
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
       61 tatcccctga ttctgtggat aaccgtagtc ggtctcatat ggatagcact gagaacgtca
      121 tcaagccctt catgcgcttc aaggtgcaca tggagggctc cgtgaacggc cacgagttcg
      181 agatcgaggg cgtgggcgag ggcaagccct acgagggcac ccagaccgcc aagctgcaag
      241 tgaccaaggg cggccccctg cccttcgcct gggacatcct gtccccccag ttcttctacg
      301 gctccaaggc gtacatcaag caccccgccg acatccccga ctacctcaag cagtccttcc
      361 ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga
      421 cccaggactc ctccctgcag gacggcaccc tcatctacca cgtgaagttc atcggcgtga
      481 acttcccctc cgacggcccc gtaatgcaga agaagactct gggctgggag ccctccactg
      541 agcgcaacta cccccgcgac ggcgtgctga agggcgagaa ccacatggcg ctgaagctga
      601 agggcggcgg ccactacctg tgtgagttca agtccatcta catggccaag aagcccgtga
      661 agctgcccgg ctaccactac gtggactaca agctcgacat cacctcccac aacgaggact
      721 acaccgtggt ggagcagtac gagcgcgccg aggcccgcca ccacctgttc cagactcacg
      781 gtatggacga attgtacaag cacgacgaat tgtaaatcct gagaccagac caataaaaaa
      841 cgcccggcgg caaccgagcg ttctgaacaa atccagatgg agttctgagg tcattactgg
      901 atctatcaac aggagtccaa gcgagctcga tatcaaatta cgccccgccc tgccactcat
      961 cgcagtactg ttgtaattca ttaagcattc tgccgacatg gaagccatca caaacggcat
     1021 gatgaacctg aatcgccagc ggcatcagca ccttgtcgcc ttgcgtataa tatttgccca
     1081 tggtgaaaac gggggcgaag aagttgtcca tattggccac gtttaaatca aaactggtga
     1141 aactcaccca gggattggct gaaacgaaaa acatattctc aataaaccct ttagggaaat
     1201 aggccaggtt ttcaccgtaa cacgccacat cttgcgaata tatgtgtaga aactgccgga
     1261 aatcgtcgtg gtattcactc cagagcgatg aaaacgtttc agtttgctca tggaaaacgg
     1321 tgtaacaagg gtgaacacta tcccatatca ccagctcacc gtctttcatt gccatacgaa
     1381 attccggatg agcattcatc aggcgggcaa gaatgtgaat aaaggccgga taaaacttgt
     1441 gcttattttt ctttacggtc tttaaaaagg ccgtaatatc cagctgaacg gtctggttat
     1501 aggtacattg agcaactgac tgaaatgcct caaaatgttc tttacgatgc cattgggata
     1561 tatcaacggt ggtatatcca gtgatttttt tctccatttt agcttcctta gctcctgaaa
     1621 atctcgataa ctcaaaaaat acgcccggta gtgatcttat ttcattatgg tgaaagttgg
     1681 aacctcttac gtgcccgatc aatcatgacc aaaatccctt aacgtgagtt ttcgttccac
     1741 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     1801 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     1861 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     1921 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     1981 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     2041 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     2101 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     2161 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     2221 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     2281 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     2341 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
//
