LOCUS       sequence-215541-        2563 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BTK assembled plasmid - used in Stage 2 assembly to target Cas9 or 
            dCas9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2563)
  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 2563)
  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: pBTK615
FEATURES             Location/Qualifiers
     source          1..2563
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_RNA        160..235
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             complement(join(690..1481,1482..1550))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1313..1332
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1551..1655)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1758..2346
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2247..2266
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 acattaaccc acagtacaga cactgcgaca acgtggcaat tcgtcgcaat accgtctcac
       61 tgaactggcc gataattgca gacgaacgct ttggcagttt attcttgaca tgtagtgagg
      121 gggctggtat aatcacatac gtctaattcc acgaggattg ttttagagct agaaatagca
      181 agttaaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt
      241 ttgaattctc tagagtcgac ctgcagaagc ttagatctat taccctgtta tccctactcg
      301 agttcatgtg cagctccatc agcatccgta atcgttaatc cgcaaataac gtaaaaaccc
      361 gcttcggcgg gtttttttat ggggggagtt tagggaaaga gcatttgtca gctgccaatg
      421 agacgacggg gtcatcacgg ctcatcatgc gccaaacaaa tgtgtgcaat acacgctcgg
      481 atgactgcat gatgaccgca ctgactgggg acagcagatc cacctaagcc tgtgagagaa
      541 gcagacaccc gacagatcaa ggcagttaac tagtgcactg cagtacagcg gccgcgatta
      601 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
      661 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
      721 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
      781 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg ggacccacgc
      841 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
      901 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
      961 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     1021 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     1081 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     1141 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     1201 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     1261 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     1321 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     1381 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     1441 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     1501 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     1561 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     1621 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     1681 gtcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     1741 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     1801 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     1861 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     1921 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     1981 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     2041 cgatagttac cggataaggc gcagcggtcg gactgaacgg ggggttcgtg cacacagccc
     2101 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     2161 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     2221 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     2281 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     2341 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     2401 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtgcggc cgcccctgaa
     2461 ttcgcatcta gatggtagag ccacaaacag ccggtacaag caacgatctc caggaccatc
     2521 tgaatcatgc gcggatgaca cgaactcacg acggcgatca cag
//
