LOCUS       sequence-226775-        3891 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  TU of the HSVtk negative marker for F2dU domesticated into pUPD2 
            with GGAG/TACT barcodes (reverse orientation). Used for gene KO with
            dual selec. According to FungalBraid modular DNA assembly for ATMT.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 3891)
  AUTHORS   Hernanz-Koers M, Gandia M, Garrigues S, Manzanares P, Yenush L, 
            Orzaez D, Marcos JF
  TITLE     FungalBraid: A GoldenBraid-based modular cloning platform for the 
            assembly and exchange of DNA elements tailored to fungal synthetic 
            biology.
  JOURNAL   Fungal Genet Biol. 2018 Jul;116:51-61. doi: 
            10.1016/j.fgb.2018.04.010. Epub 2018 Apr 20.
  PUBMED    29680684
REFERENCE   2  (bases 1 to 3891)
  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: "Fungal 
            Genet Biol."; date: "2018-07"; volume: "116:51-61. doi"; pages: " 
            10.1016/j.fgb.2018.04.010. Epub 2018 Apr 20"
COMMENT     Sequence Label: FB013
FEATURES             Location/Qualifiers
     source          1..3891
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(285..1415)
                     /codon_start=1
                     /product="herpes simplex virus thymidine kinase"
                     /label=HSV TK
                     /note="confers sensitivity to nucleoside analogs such as 
                     acyclovir or ganciclovir"
                     /translation="MASYPCHQHASAFDQAARSRGHSNRRTALRPRRQQEATEVRLEQK
                     MPTLLRVYIDGPHGMGKTTTTQLLVALGSRDDIVYVPEPMTYWQVLGASETIANIYTTQ
                     HRLDQGEISAGDAAVVMTSAQITMGMPYAVTDAVLAPHIGGEAGSSHAPPPALTLIFDR
                     HPIAALLCYPAARYLMGSMTPQAVLAFVALIPPTLPGTNIVLGALPEDRHIDRLAKRQR
                     PGERLDLAMLAAIRRVYGLLANTVRYLQGGGSWREDWGQLSGTAVPPQGAEPQSNAGPR
                     PHIGDTLFTLFRAPELLAPNGDLYNVFAWALDVLAKRLRPMHVFILDYDQSPAGCRDAL
                     LQLTSGMVQTHVTTPGSIPTICDLARTFAREMGEAN"
     terminator      1828..1885
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     primer_bind     complement(1909..1926)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2080..2668)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2160..2179)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2851..2945)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2966..3625)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3540..3559
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     3787..3805
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(3809..3852)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    3855..3876
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
ORIGIN
        1 ggagtcatca tgcaacatgc atgtactgtc tgatgtatta agagtatagg ggtcaacaac
       61 tagagtgtct gacaggtata tctagcagtt tgatttgacc attcttttat taaaacactg
      121 taacccgcaa cgaagaactc ctccgcttca agtcaagaat gcgcatatgt gcgtaaaatc
      181 gacaagcccg gcggcattcc accgagcagg gtattgcctt tggcacttta cgtgcgcaga
      241 ggagcctgaa tgttgagtgg aatgatgcat gcctgcacct cgactcagtt agcctccccc
      301 atctcgcggg caaacgtgcg cgccaggtcg cagatcgtcg gtatggagcc gggggtggtg
      361 acgtgggtct ggaccatccc ggaggtaagt tgcagcaggg cgtcccggca gccggcgggc
      421 gattggtcgt aatccaggat aaagacgtgc atgggacgga ggcgtttggc caagacgtcc
      481 aaggcccagg caaacacgtt gtacaggtcg ccgttggggg ccagcaactc gggggcccga
      541 aacagggtaa ataacgtgtc cccgatatgg ggtcgtgggc ccgcgttgct ctggggctcg
      601 gcaccctggg gcggcacggc cgtccccgaa agctgtcccc aatcctcccg ccacgacccg
      661 ccgccctgga gataccgcac cgtattggca agcagcccgt aaacgcggcg aatcgcggcc
      721 agcatagcca ggtcaagccg ctcgccgggg cgctggcgtt tggccaggcg gtcgatgtgt
      781 ctgtcctctg gaagggcccc caacacgatg tttgtgccgg gcaaggtcgg cgggatgagg
      841 gccacgaacg ccagcacggc ctggggggtc atgctgccca taaggtatcg cgcggccggg
      901 tagcacagga gggcggcgat gggatggcgg tcgaagatga gggtgagggc cgggggcggg
      961 gcatgtgagc tgccagcctc ccccccgata tgaggagcca gaacggcgtc ggtcacggca
     1021 taaggcatgc ccattgttat ctgggcgctt gtcattacca ccgccgcgtc cccggccgaa
     1081 atctcaccct ggtcgaggcg gtgttgtgtg gtgtagatgt tcgcgattgt ctcggaagcc
     1141 cccagcacct gccagtaagt catcggctcg ggtacgtaga cgatgtcgtc gcgcgaaccc
     1201 agggccacca gcagttgcgt ggtggtggtt ttccccatcc cgtgaggacc gtctatataa
     1261 acccgcagta gcgtgggcat tttctgctcc aggcggactt ccgtggcttc ttgctgccgg
     1321 cgagggcgca acgccgtacg tcggttgcta tggccgcgag aacgcgcagc ctggtcgaac
     1381 gcagacgcgt gttgatggca ggggtacgaa gccatggatc tgaagattgg gttcctcttt
     1441 gagagagtta atagacttga attgaggaat ggaaatcggg ctgagctcac tgctttatat
     1501 gtgagaaaag gtgatgagca ggtggtaaga ggcagatttt tgtggtcggt gctgtgtttt
     1561 gttgggtttg agctaggtgg ggtgagggag gggcaagcta aatagctacc agccctgagg
     1621 acgtcacaga gttttagtag ttctttttct gccatcggac atgcagctca cgcctagacc
     1681 caattacacc ctttgcgccc tcccacacat ccggccggtt ggatcattgt cagatacggc
     1741 agagaaatcg caacctcggc caagggtagc cacgattcga agccgctact tgagaccact
     1801 catcgcgact agtagcggcc gctgcagtcc ggcaaaaaag ggcaaggtgt caccaccctg
     1861 ccctttttct ttaaaaccga aaagattact tcgcgttatg caggcttcct cgctcactga
     1921 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     1981 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     2041 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccacaggc tccgcccccc
     2101 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     2161 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     2221 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     2281 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     2341 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     2401 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     2461 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     2521 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     2581 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     2641 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     2701 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     2761 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     2821 gtaaacttgg tctgacagct cgaggcttgg attctcacca ataaaaaacg cccggcggca
     2881 accgagcgtt ctgaacaaat ccagatggag ttctgaggtc attactggat ctatcaacag
     2941 gagtccaagc gagctcgata tcaaattacg ccccgccctg ccactcgtcg cagtactgtt
     3001 gtaattcatt aagcattctg ccgacatgga agccatcaca aacggcatga tgaacctgaa
     3061 tcgccagcgg catcagcacc ttgtcgcctt gcgtataata tttgcccatg gtgaaaacgg
     3121 gggcgaagaa gttgtccata ttggccacgt ttaaatcaaa actggtgaaa ctcacccagg
     3181 gattggctga tacgaaaaac atattctcaa taaacccttt agggaaatag gccaggtttt
     3241 caccgtaaca cgccacatct tgcgaatata tgtgtagaaa ctgccggaaa tcgtcgtggt
     3301 attcactcca gagtgatgaa aacgtttcag tttgctcatg gaaaacggtg taacaagggt
     3361 gaacactatc ccatatcacc agctcaccgt ctttcattgc catacgaaat tccggatgag
     3421 cattcatcag gcgggcaaga atgtgaataa aggccggata aaacttgtgc ttatttttct
     3481 ttacggtctt taaaaaggcc gtaatatcca gctgaacggt ctggttatag gtacattgag
     3541 caactgactg aaatgcctca aaatgttctt tacgatgcca ttgggatata tcaacggtgg
     3601 tatatccagt gatttttttc tccattttag cttccttagc tcctgaaaat ctcgataact
     3661 caaaaaatac gcccggtagt gatcttattt cattatggtg aaagttggaa cctcttacgt
     3721 gcccgatcaa ctcgagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
     3781 tataaaaata ggcgtatcac gaggcagaat ttcagataaa aaaaatcctt agctttcgct
     3841 aaggatgatt tctggaattc gcggccgctt ctagagcgat gagtggtctc g
//
