LOCUS       sequence-208048-        2673 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BTK Type 3 coding sequence for golden gate assembly and homologous 
            recombination.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2673)
  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 2673)
  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: pBTK229
FEATURES             Location/Qualifiers
     source          1..2673
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     205..224
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(462..1277)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(462..1277)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     1185..1204
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1269..1288
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     terminator      complement(1443..1477)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(1548..2207)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2122..2141
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      join(2389..2673,1..304)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
       61 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
      121 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
      181 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
      241 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
      301 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
      361 catgttcttt cctgcgttat cccctgattc tgtggataac cgtagtcggt ctcatatgta
      421 taccgagcgg ccgcgtgtta caaccaatta accaattctg attagaaaaa ctcatcgagc
      481 atcaaatgaa actgcaattt attcatatca ggattatcaa taccatattt ttgaaaaagc
      541 cgtttctgta atgaaggaga aaactcaccg aggcagttcc ataggatggc aagatcctgg
      601 tatcggtctg cgattccgac tcgtccaaca tcaatacaac ctattaattt cccctcgtca
      661 aaaataaggt tatcaagtga gaaatcacca tgagtgacga ctgaatccgg tgagaatggc
      721 aaaagcttat gcatttcttt ccagacttgt tcaacaggcc agccattacg ctcgtcatca
      781 aaatcactcg catcaaccaa accgttattc attcgtgatt gcgcctgagc gaggcgaaat
      841 acgcgatcgc tgttaaaagg acaattacaa acaggaatcg aatgcaaccg gcgcaggaac
      901 actgccagcg catcaacaat attttcacct gaatcaggat attcttctaa tacctggaat
      961 gctgttttcc cggggatcgc agtggtgagt aaccatgcat catcaggagt acggataaaa
     1021 tgcttgatgg tcggaagagg cataaattcc gtcagccagt ttagtctgac catctcatct
     1081 gtaacatcat tggcaacgct acctttgcca tgtttcagaa acaactctgg cgcatcgggc
     1141 ttcccataca atcgatagat tgtcgcacct gattgcccga cattatcgcg agcccattta
     1201 tacccatata aatcagcatc catgttggaa tttaatcgcg gcctggagca agacgtttcc
     1261 cgttgaatat ggctcataac accccttgta ttactgttta tgtaagcaga cagttttatt
     1321 gttcatgatg atatattttt atcttgtgca atgtaacatc agagattttg agacacaacg
     1381 tggctttgtt gaataaatcg aacttttgct gagttgaagg atcagatcct gagaccagac
     1441 caataaaaaa cgcccggcgg caaccgagcg ttctgaacaa atccagatgg agttctgagg
     1501 tcattactgg atctatcaac aggagtccaa gcgagctcga tatcaaatta cgccccgccc
     1561 tgccactcat cgcagtactg ttgtaattca ttaagcattc tgccgacatg gaagccatca
     1621 caaacggcat gatgaacctg aatcgccagc ggcatcagca ccttgtcgcc ttgcgtataa
     1681 tatttgccca tggtgaaaac gggggcgaag aagttgtcca tattggccac gtttaaatca
     1741 aaactggtga aactcaccca gggattggct gaaacgaaaa acatattctc aataaaccct
     1801 ttagggaaat aggccaggtt ttcaccgtaa cacgccacat cttgcgaata tatgtgtaga
     1861 aactgccgga aatcgtcgtg gtattcactc cagagcgatg aaaacgtttc agtttgctca
     1921 tggaaaacgg tgtaacaagg gtgaacacta tcccatatca ccagctcacc gtctttcatt
     1981 gccatacgaa attccggatg agcattcatc aggcgggcaa gaatgtgaat aaaggccgga
     2041 taaaacttgt gcttattttt ctttacggtc tttaaaaagg ccgtaatatc cagctgaacg
     2101 gtctggttat aggtacattg agcaactgac tgaaatgcct caaaatgttc tttacgatgc
     2161 cattgggata tatcaacggt ggtatatcca gtgatttttt tctccatttt agcttcctta
     2221 gctcctgaaa atctcgataa ctcaaaaaat acgcccggta gtgatcttat ttcattatgg
     2281 tgaaagttgg aacctcttac gtgcccgatc aatcatgacc aaaatccctt aacgtgagtt
     2341 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     2401 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     2461 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     2521 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     2581 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     2641 taagtcgtgt cttaccgggt tggactcaag acg
//
