LOCUS       sequence-468939-        2806 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BTK Type 2 antibiotic resistance cassette for Golden Gate assembly.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2806)
  AUTHORS   Lariviere PJ, Zuberi Ashraf AHM, Leonard SP, Miller LG, Moran NA, 
            Barrick JE
  TITLE     Single-step genome engineering in the bee gut symbiont Snodgrassella
            alvi
  JOURNAL   bioRxiv 2023.09.19.558440
REFERENCE   2  (bases 1 to 2806)
  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"; journalName: "bioRxiv 
            2023.09.19.558440"
COMMENT     Sequence Label: pBTK1052
FEATURES             Location/Qualifiers
     source          1..2806
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     7..26
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     91..110
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     terminator      complement(320..354)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(425..1084)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     999..1018
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      1266..1854
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1755..1774
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2090..2806,1..99))
                     /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(join(2090..2806,1..99))
                     /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"
ORIGIN
        1 gacattatcg cgagcccatt tatacccata taaatcagca tccatgttgg aatttaatcg
       61 cggcctggag caagacgttt cccgttgaat atggctcata acaccccttg tattactgtt
      121 tatgtaagca gacagtttta ttgttcatga tgatatattt ttatcttgtg caatgtaaca
      181 tcagagattt tgagacacaa cgtggctttg ttgaataaat cgaacttttg ctgagttgaa
      241 ggatcagtat gtgagaccgg aaacacagaa aaaagcccac acctgacagt gcgggctttt
      301 tttttcgacc aaaggaccaa taaaaaacgc ccggcggcaa ccgagcgttc tgaacaaatc
      361 cagatggagt tctgaggtca ttactggatc tatcaacagg agtccaagcg agctcgatat
      421 caaattacgc cccgccctgc cactcatcgc agtactgttg taattcatta agcattctgc
      481 cgacatggaa gccatcacaa acggcatgat gaacctgaat cgccagcggc atcagcacct
      541 tgtcgccttg cgtataatat ttgcccatgg tgaaaacggg ggcgaagaag ttgtccatat
      601 tggccacgtt taaatcaaaa ctggtgaaac tcacccaggg attggctgaa acgaaaaaca
      661 tattctcaat aaacccttta gggaaatagg ccaggttttc accgtaacac gccacatctt
      721 gcgaatatat gtgtagaaac tgccggaaat cgtcgtggta ttcactccag agcgatgaaa
      781 acgtttcagt ttgctcatgg aaaacggtgt aacaagggtg aacactatcc catatcacca
      841 gctcaccgtc tttcattgcc atacgaaatt ccggatgagc attcatcagg cgggcaagaa
      901 tgtgaataaa ggccggataa aacttgtgct tatttttctt tacggtcttt aaaaaggccg
      961 taatatccag ctgaacggtc tggttatagg tacattgagc aactgactga aatgcctcaa
     1021 aatgttcttt acgatgccat tgggatatat caacggtggt atatccagtg atttttttct
     1081 ccattttagc ttccttagct cctgaaaatc tcgataactc aaaaaatacg cccggtagtg
     1141 atcttatttc attatggtga aagttggaac ctcttacgtg cccgatcaat catgaccaaa
     1201 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     1261 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     1321 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     1381 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     1441 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     1501 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     1561 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     1621 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     1681 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     1741 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     1801 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     1861 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     1921 cctgcgttat cccctgattc tgtggataac cgtacgtttg agaagagaaa agaaaaccgc
     1981 cgatcctgtc caccgcatta ctgcaaggta gtggacaaga ccggcggtct taagtttttt
     2041 ggctgaaagt cggtctcaaa cggtgttaca accaattaac caattctgat tagaaaaact
     2101 catcgagcat caaatgaaac tgcaatttat tcatatcagg attatcaata ccatattttt
     2161 gaaaaagccg tttctgtaat gaaggagaaa actcaccgag gcagttccat aggatggcaa
     2221 gatcctggta tcggtctgcg attccgactc gtccaacatc aatacaacct attaatttcc
     2281 cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg agtgacgact gaatccggtg
     2341 agaatggcaa aagcttatgc atttctttcc agacttgttc aacaggccag ccattacgct
     2401 cgtcatcaaa atcactcgca tcaaccaaac cgttattcat tcgtgattgc gcctgagcga
     2461 ggcgaaatac gcgatcgctg ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc
     2521 gcaggaacac tgccagcgca tcaacaatat tttcacctga atcaggatat tcttctaata
     2581 cctggaatgc tgttttcccg gggatcgcag tggtgagtaa ccatgcatca tcaggagtac
     2641 ggataaaatg cttgatggtc ggaagaggca taaattccgt cagccagttt agtctgacca
     2701 tctcatctgt aacatcattg gcaacgctac ctttgccatg tttcagaaac aactctggcg
     2761 catcgggctt cccatacaat cgatagattg tcgcacctga ttgccc
//
