LOCUS       sequence-233535-        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"
     rep_origin      complement(150..738)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(230..249)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             920..1579
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(986..1005)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1650..1684
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     primer_bind     complement(1839..1858)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1850..2665
                     /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             1850..2665
                     /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     complement(1923..1942)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 ggttaattgg ttgtaacacg cggccgctcg gtatacatat gagaccgact acggttatcc
       61 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
      121 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
      181 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
      241 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
      301 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
      361 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
      421 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
      481 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
      541 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
      601 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
      661 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
      721 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
      781 aacgaaaact cacgttaagg gattttggtc atgattgatc gggcacgtaa gaggttccaa
      841 ctttcaccat aatgaaataa gatcactacc gggcgtattt tttgagttat cgagattttc
      901 aggagctaag gaagctaaaa tggagaaaaa aatcactgga tataccaccg ttgatatatc
      961 ccaatggcat cgtaaagaac attttgaggc atttcagtca gttgctcaat gtacctataa
     1021 ccagaccgtt cagctggata ttacggcctt tttaaagacc gtaaagaaaa ataagcacaa
     1081 gttttatccg gcctttattc acattcttgc ccgcctgatg aatgctcatc cggaatttcg
     1141 tatggcaatg aaagacggtg agctggtgat atgggatagt gttcaccctt gttacaccgt
     1201 tttccatgag caaactgaaa cgttttcatc gctctggagt gaataccacg acgatttccg
     1261 gcagtttcta cacatatatt cgcaagatgt ggcgtgttac ggtgaaaacc tggcctattt
     1321 ccctaaaggg tttattgaga atatgttttt cgtttcagcc aatccctggg tgagtttcac
     1381 cagttttgat ttaaacgtgg ccaatatgga caacttcttc gcccccgttt tcaccatggg
     1441 caaatattat acgcaaggcg acaaggtgct gatgccgctg gcgattcagg ttcatcatgc
     1501 cgtttgtgat ggcttccatg tcggcagaat gcttaatgaa ttacaacagt actgcgatga
     1561 gtggcagggc ggggcgtaat ttgatatcga gctcgcttgg actcctgttg atagatccag
     1621 taatgacctc agaactccat ctggatttgt tcagaacgct cggttgccgc cgggcgtttt
     1681 ttattggtct ggtctcagga tctgatcctt caactcagca aaagttcgat ttattcaaca
     1741 aagccacgtt gtgtctcaaa atctctgatg ttacattgca caagataaaa atatatcatc
     1801 atgaacaata aaactgtctg cttacataaa cagtaataca aggggtgtta tgagccatat
     1861 tcaacgggaa acgtcttgct ccaggccgcg attaaattcc aacatggatg ctgatttata
     1921 tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgattgta
     1981 tgggaagccc gatgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga
     2041 tgttacagat gagatggtca gactaaactg gctgacggaa tttatgcctc ttccgaccat
     2101 caagcatttt atccgtactc ctgatgatgc atggttactc accactgcga tccccgggaa
     2161 aacagcattc caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct
     2221 ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt ttaacagcga
     2281 tcgcgtattt cgcctcgctc aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag
     2341 tgattttgat gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcataa
     2401 gcttttgcca ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct
     2461 tatttttgac gaggggaaat taataggttg tattgatgtt ggacgagtcg gaatcgcaga
     2521 ccgataccag gatcttgcca tcctatggaa ctgcctcggt gagttttctc cttcattaca
     2581 gaaacggctt tttcaaaaat atggtattga taatcctgat atgaataaat tgcagtttca
     2641 tttgatgctc gatgagtttt tctaatcaga att
//
