LOCUS       sequence-42231-e        2395 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  natural linear DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 2395)
  TITLE     Plasmid for gene disruption
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 2395)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pTN186
FEATURES             Location/Qualifiers
     source          1..2395
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     13..29
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     14..30
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(48..81)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        145..492
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     435..455
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             512..1537
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     polyA_signal    1579..1755
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     protein_bind    complement(2244..2277)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(2369..2389)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2369..2387)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 ccgggccccc cctcgaggtc gacggtatcg ataagcttga tatcgaaata acttcgtata
       61 gcatacatta tacgaagtta tattcctgca gcccccctac tccaaaaatg tcaaagatac
      121 agtctcagaa gaccaaaggg ctattgacac ttttcaacaa agggtatata atcgggaaga
      181 ctcctcggat tccattgccc agctatctgt cacttcatcg aaaggacagt agaaaaggaa
      241 ggtggctcct acaaatgcca tcattgcgat aaaggaaagg ctatcattca agatgcctct
      301 gccgacagtg gtcccaaaga tggaccccca cccacgagga gcatcgtgga aaaagaagac
      361 gttccaacca cgtcttcaaa gcaagtggat tgatgtgaca tctccactga cgtaagggat
      421 gacgcacaat cccactatcc ttcgcaagac ccttcctcta tataaggaag ttcatttcat
      481 ttggagagga cagcccaagc aaggaggtga tatgaagaag cctgaactca ccgcgacgtc
      541 tgtcgagaag tttctgatcg aaaagttcga cagcgtctcc gacctgatgc agctctcgga
      601 gggcgaagaa tctcgtgctt tcagcttcga tgtaggaggg cgtggatatg tcctgcgggt
      661 aaatagctgc gccgatggtt tctacaaaga tcgttatgtt tatcggcact ttgcatcggc
      721 cgcgctcccg attccggaag tgcttgacat tggagagttc agcgagagcc tgacctattg
      781 catctcccgc cgtgcacagg gtgtcacgtt gcaagacctg cctgaaaccg aactgcccgc
      841 tgttcttcag ccggtcgcgg aggctatgga tgcgatcgct gcggccgatc ttagccagac
      901 gagcgggttc ggcccattcg gaccgcaagg aatcggtcaa tacactacat ggcgtgattt
      961 catatgcgcg attgctgatc cccatgtgta tcactggcaa actgtgatgg acgacaccgt
     1021 cagtgcgtcc gtcgcgcagg ctctcgatga gctgatgctt tgggccgagg actgccccga
     1081 agtccggcac ctcgtgcacg cggatttcgg ctccaacaat gtcctgacgg acaatggccg
     1141 cataacagcg gtcattgact ggagcgaggc gatgttcggg gattcccaat acgaggtcgc
     1201 caacatcttc ttctggaggc cgtggttggc ttgtatggag cagcagacgc gctacttcga
     1261 gcggaggcat ccggagcttg caggatcgcc gcggctccgg gcgtatatgc tccgcattgg
     1321 tcttgaccaa ctctatcaga gcttggttga cggcaatttc gatgatgcag cttgggcgca
     1381 gggtcgatgc gacgcaatcg tccgatccgg agccgggact gtcgggcgta cacaaatcgc
     1441 ccgcagaagc gcggccgtct ggaccgatgg ctgtgtagaa gtactcgccg atagtggaaa
     1501 ccgacgcccc agcactcgtc ctagggcaaa ggaatagtga aatcaccagt ctctctctac
     1561 aaatctatct ctctctattt tctccataaa taatgtgtga gtagtttccc gataagggaa
     1621 attagggttc ttatagggtt tcgctcatgt gttgagcata taagaaaccc ttagtatgta
     1681 tttgtatttg taaaatactt ctatcaataa aatttctaat tcctaaaacc aaaatccagt
     1741 actaaaatcc agatctccta aagtccctat agatctttgt cgtgaatata aaccagacac
     1801 gagacgacta aacctggagc ccagacgccg ttcgaagcta gaagtaccgc ttaggcagga
     1861 ggccgttagg gaaaagatgc taaggcaggg ttggttacgt tgactccccc gtaggtttgg
     1921 tttaaatatg atgaagtgga cggaaggaag gaggaagaca aggaaggata aggttgcagg
     1981 ccctgtgcaa ggtaagaaga tggaaatttg atagaggtac gctactatac ttatactata
     2041 cgctaaggga atgcttgtat ttatacccta taccccctaa taacccctta tcaatttaag
     2101 aaataatccg cataagcccc cgcttaaaaa ttggtatcag ggccatgaat aggtctatga
     2161 ccaaaactca agaggataaa acctcaccaa aatacgaaag agttcttaac tctaaagata
     2221 aaagatcttt caagatcaaa actataactt cgtatagcat acattatacg aagttatccc
     2281 tcacaccggt gacggggatc gcatgcccgg gtctagacat atggatccta gagcggccgc
     2341 caccgcggtg gagctccagc ttttgttccc tttagtgagg gttaattgcg cgctg
//
