LOCUS       sequence-419119-        2748 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BTK Type 4 antibiotic resistance cassette for Golden Gate assembly.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2748)
  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 2748)
  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: pBTK1056
FEATURES             Location/Qualifiers
     source          1..2748
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(15..49)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(120..779)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     694..713
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      961..1549
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1450..1469
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(1834..1853)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1845..2660
                     /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             1845..2660
                     /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(1918..1937)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 cgaccaaagg accaataaaa aacgcccggc ggcaaccgag cgttctgaac aaatccagat
       61 ggagttctga ggtcattact ggatctatca acaggagtcc aagcgagctc gatatcaaat
      121 tacgccccgc cctgccactc atcgcagtac tgttgtaatt cattaagcat tctgccgaca
      181 tggaagccat cacaaacggc atgatgaacc tgaatcgcca gcggcatcag caccttgtcg
      241 ccttgcgtat aatatttgcc catggtgaaa acgggggcga agaagttgtc catattggcc
      301 acgtttaaat caaaactggt gaaactcacc cagggattgg ctgaaacgaa aaacatattc
      361 tcaataaacc ctttagggaa ataggccagg ttttcaccgt aacacgccac atcttgcgaa
      421 tatatgtgta gaaactgccg gaaatcgtcg tggtattcac tccagagcga tgaaaacgtt
      481 tcagtttgct catggaaaac ggtgtaacaa gggtgaacac tatcccatat caccagctca
      541 ccgtctttca ttgccatacg aaattccgga tgagcattca tcaggcgggc aagaatgtga
      601 ataaaggccg gataaaactt gtgcttattt ttctttacgg tctttaaaaa ggccgtaata
      661 tccagctgaa cggtctggtt ataggtacat tgagcaactg actgaaatgc ctcaaaatgt
      721 tctttacgat gccattggga tatatcaacg gtggtatatc cagtgatttt tttctccatt
      781 ttagcttcct tagctcctga aaatctcgat aactcaaaaa atacgcccgg tagtgatctt
      841 atttcattat ggtgaaagtt ggaacctctt acgtgcccga tcaatcatga ccaaaatccc
      901 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
      961 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     1021 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     1081 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     1141 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     1201 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     1261 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     1321 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     1381 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     1441 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     1501 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     1561 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     1621 gttatcccct gattctgtgg ataaccgtac gtttgagaag agaaaagaaa accgccgatc
     1681 ctgtccaccg cattactgca aggtagtgga caagaccggc ggtcttaagt tttttggctg
     1741 aaagtcggtc tcaatccctc tgatgttaca ttgcacaaga taaaaatata tcatcatgaa
     1801 caataaaact gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac
     1861 gggaaacgtc ttgctccagg ccgcgattaa attccaacat ggatgctgat ttatatgggt
     1921 ataaatgggc tcgcgataat gtcgggcaat caggtgcgac aatctatcga ttgtatggga
     1981 agcccgatgc gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta
     2041 cagatgagat ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc
     2101 attttatccg tactcctgat gatgcatggt tactcaccac tgcgatcccc gggaaaacag
     2161 cattccaggt attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag
     2221 tgttcctgcg ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg
     2281 tatttcgcct cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt
     2341 ttgatgacga gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataagcttt
     2401 tgccattctc accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt
     2461 ttgacgaggg gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat
     2521 accaggatct tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac
     2581 ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga
     2641 tgctcgatga gtttttctaa tcagaattgg ttaattggtt gtaacacgcg gctgtgagac
     2701 cggaaacaca gaaaaaagcc cacacctgac agtgcgggct tttttttt
//
