LOCUS       sequence-474009-        3118 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BsaI compatible kanamycin resistance cassette.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3118)
  AUTHORS   VanDieren AJ, Barrick JE
  TITLE     UltraCAST: A Flexible All-In-One Suicide Vector for Modifying 
            Bacterial Genomes Using a CRISPR-Associated Transposon.
  JOURNAL   MicroPubl Biol. 2025 Aug 2;2025:10.17912/micropub.biology.001721. 
            doi: 10.17912/micropub.biology.001721. eCollection 2025.
  PUBMED    40827214
REFERENCE   2  (bases 1 to 3118)
  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: "MicroPubl 
            Biol."; date: "2025-08-2"; volume: 
            "2025:10.17912/micropub.biology.001721. doi"; pages: " 
            10.17912/micropub.biology.001721. eCollection 2025"
COMMENT     Sequence Label: pBTK1062
FEATURES             Location/Qualifiers
     source          1..3118
                     /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"
     protein_bind    complement(1777..1824)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             complement(1837..2631)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1948..1967)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2558..2577
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     protein_bind    complement(2999..3032)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
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 tcaaacgtgt aggctggagc tgcttcgaag ttcctatact ttctagagaa
     1801 taggaacttc ggaataggaa cttcaagatc cccttattag aagaactcgt caagaaggcg
     1861 atagaaggcg atgcgctgcg aatcgggagc ggcgataccg taaagcacga ggaagcggtc
     1921 agcccattcg ccgccaagct cttcagcaat atcacgggta gccaacgcta tgtcctgata
     1981 gcggtccgcc acacccagcc ggccacagtc gatgaatcca gaaaagcggc cattttccac
     2041 catgatattc ggcaagcagg catcgccatg ggtcacgacg agatcctcgc cgtcgggcat
     2101 gcgcgccttg agcctggcga acagttcggc tggcgcgagc ccctgatgct cttcgtccag
     2161 atcatcctga tcgacaagac cggcttccat ccgagtacgt gctcgctcga tgcgatgttt
     2221 cgcttggtgg tcgaatgggc aggtagccgg atcaagcgta tgcagccgcc gcattgcatc
     2281 agccatgatg gatactttct cggcaggagc aaggtgagat gacaggagat cctgccccgg
     2341 cacttcgccc aatagcagcc agtcccttcc cgcttcagtg acaacgtcga gcacagctgc
     2401 gcaaggaacg cccgtcgtgg ccagccacga tagccgcgct gcctcgtcct gcagttcatt
     2461 cagggcaccg gacaggtcgg tcttgacaaa aagaaccggg cgcccctgcg ctgacagccg
     2521 gaacacggcg gcatcagagc agccgattgt ctgttgtgcc cagtcatagc cgaatagcct
     2581 ctccacccaa gcggccggag aacctgcgtg caatccatct tgttcaatca tgcgaaacga
     2641 tcctcatcct gtctcttgat cagatcttga tcccctgcgc catcagatcc ttggcggcaa
     2701 gaaagccatc cagtttactt tgcagggctt cccaacctta ccagagggcg ccccagctgg
     2761 caattccggt tcgcttgctg tccataaaac cgcccagtct agctatcgcc atgtaagccc
     2821 actgcaagct acctgctttc tctttgcgct tgcgttttcc cttgtccaga tagcccagta
     2881 gctgacattc atccggggtc agcaccgttt ctgcggactg gctttctacg tgttccgctt
     2941 cctttagcag cccttgcgcc ctgagtgctt gcggcagcgt gagcttcaaa agcgctctga
     3001 agttcctata ctttctagag aataggaact tcgaactgca ggtcgacgga tccccggaat
     3061 gctgtgagac cggaaacaca gaaaaaagcc cacacctgac agtgcgggct tttttttt
//
