LOCUS       sequence-42229-e        2500 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 2500)
  AUTHORS   Sakakibara K, Nishiyama T, Deguchi H, Hasebe M
  TITLE     Class 1 KNOX genes are not involved in shoot development in the moss
            Physcomitrella patens but do function in sporophyte development.
  JOURNAL   Evol Dev. 2008 Sep-Oct;10(5):555-66.
  PUBMED    18803774
REFERENCE   2  (bases 1 to 2500)
  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: "Evol Dev. 
            2008 Sep-Oct"; volume: "10(5)"; pages: "555-66"
COMMENT     Sequence Label: pTN182
FEATURES             Location/Qualifiers
     source          1..2500
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     171..193
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     185..202
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     186..202
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     212..231
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        212..230
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     255..271
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     256..272
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(294..327)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        382..729
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     672..692
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             750..1544
                     /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(804..823)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1414..1433
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1614..1790
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     protein_bind    complement(2279..2312)
                     /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(2404..2424)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2404..2422)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2443..2459)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2443..2459)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2456..2478)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2467..2483
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
ORIGIN
        1 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcccatt cgccattcag
       61 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
      121 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
      181 acgttgtaaa acgacggcca gtgagcgcgc gtaatacgac tcactatagg gcgaattggg
      241 taccgggccc cccctcgagg tcgacggtat cgataagctt gatatcgaat tccataactt
      301 cgtatagcat acattatacg aagttatgcc cccctactcc aaaaatgtca aagatacagt
      361 ctcagaagac caaagggcta ttgacacttt tcaacaaagg gtatataatc gggaagactc
      421 ctcggattcc attgcccagc tatctgtcac ttcatcgaaa ggacagtaga aaaggaaggt
      481 ggctcctaca aatgccatca ttgcgataaa ggaaaggcta tcattcaaga tgcctctgcc
      541 gacagtggtc ccaaagatgg acccccaccc acgaggagca tcgtggaaaa agaagacgtt
      601 ccaaccacgt cttcaaagca agtggattga tgtgacatct ccactgacgt aagggatgac
      661 gcacaatccc actatccttc gcaagaccct tcctctatat aaggaagttc atttcatttg
      721 gagaggacag cccaagcaag gaggtgagca tgattgaaca agacggattg cacgcaggtt
      781 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
      841 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
      901 ccgacctgtc cggtgccctg aatgaactcc aagacgaggc agcgcggcta tcgtggctgg
      961 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     1021 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     1081 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     1141 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     1201 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     1261 tcgccaggct caaggcgcgg atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     1321 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     1381 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     1441 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     1501 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     1561 cggacggtac gctgaaatca ccagtctctc tctacaaatc tatctctctc tattttctcc
     1621 ataaataatg tgtgagtagt ttcccgataa gggaaattag ggttcttata gggtttcgct
     1681 catgtgttga gcatataaga aacccttagt atgtatttgt atttgtaaaa tacttctatc
     1741 aataaaattt ctaattccta aaaccaaaat ccagtactaa aatccagatc tcctaaagtc
     1801 cctatagatc tttgtcgtga atataaacca gacacgagac gactaaacct ggagcccaga
     1861 cgccgttcga agctagaagt accgcttagg caggaggccg ttagggaaaa gatgctaagg
     1921 cagggttggt tacgttgact cccccgtagg tttggtttaa atatgatgaa gtggacggaa
     1981 ggaaggagga agacaaggaa ggataaggtt gcaggccctg tgcaaggtaa gaagatggaa
     2041 atttgataga ggtacgctac tatacttata ctatacgcta agggaatgct tgtatttata
     2101 ccctataccc cctaataacc ccttatcaat ttaagaaata atccgcataa gcccccgctt
     2161 aaaaattggt atcagggcca tgaataggtc tatgaccaaa actcaagagg ataaaacctc
     2221 accaaaatac gaaagagttc ttaactctaa agataaaaga tctttcaaga tcaaaactat
     2281 aacttcgtat agcatacatt atacgaagtt atccctcaca ccggtgacgg ggatcgcatg
     2341 cccgggtcta gacatatgga tcctagagcg gccgccaccg cggtggagct ccagcttttg
     2401 ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt
     2461 gtgaaattgt tatccgctca caattccaca caacatacga
//
