LOCUS       sequence-237145-        3891 bp    DNA     circular SYN 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      synthetic DNA construct
  ORGANISM  synthetic DNA construct
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="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(1..44)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    47..68
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     CDS             complement(368..1498)
                     /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      1911..1968
                     /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(1992..2009)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2163..2751)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2243..2262)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2934..3028)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3049..3708)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3623..3642
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     3870..3888
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 atttcagata aaaaaaatcc ttagctttcg ctaaggatga tttctggaat tcgcggccgc
       61 ttctagagcg atgagtggtc tcgggagtca tcatgcaaca tgcatgtact gtctgatgta
      121 ttaagagtat aggggtcaac aactagagtg tctgacaggt atatctagca gtttgatttg
      181 accattcttt tattaaaaca ctgtaacccg caacgaagaa ctcctccgct tcaagtcaag
      241 aatgcgcata tgtgcgtaaa atcgacaagc ccggcggcat tccaccgagc agggtattgc
      301 ctttggcact ttacgtgcgc agaggagcct gaatgttgag tggaatgatg catgcctgca
      361 cctcgactca gttagcctcc cccatctcgc gggcaaacgt gcgcgccagg tcgcagatcg
      421 tcggtatgga gccgggggtg gtgacgtggg tctggaccat cccggaggta agttgcagca
      481 gggcgtcccg gcagccggcg ggcgattggt cgtaatccag gataaagacg tgcatgggac
      541 ggaggcgttt ggccaagacg tccaaggccc aggcaaacac gttgtacagg tcgccgttgg
      601 gggccagcaa ctcgggggcc cgaaacaggg taaataacgt gtccccgata tggggtcgtg
      661 ggcccgcgtt gctctggggc tcggcaccct ggggcggcac ggccgtcccc gaaagctgtc
      721 cccaatcctc ccgccacgac ccgccgccct ggagataccg caccgtattg gcaagcagcc
      781 cgtaaacgcg gcgaatcgcg gccagcatag ccaggtcaag ccgctcgccg gggcgctggc
      841 gtttggccag gcggtcgatg tgtctgtcct ctggaagggc ccccaacacg atgtttgtgc
      901 cgggcaaggt cggcgggatg agggccacga acgccagcac ggcctggggg gtcatgctgc
      961 ccataaggta tcgcgcggcc gggtagcaca ggagggcggc gatgggatgg cggtcgaaga
     1021 tgagggtgag ggccgggggc ggggcatgtg agctgccagc ctcccccccg atatgaggag
     1081 ccagaacggc gtcggtcacg gcataaggca tgcccattgt tatctgggcg cttgtcatta
     1141 ccaccgccgc gtccccggcc gaaatctcac cctggtcgag gcggtgttgt gtggtgtaga
     1201 tgttcgcgat tgtctcggaa gcccccagca cctgccagta agtcatcggc tcgggtacgt
     1261 agacgatgtc gtcgcgcgaa cccagggcca ccagcagttg cgtggtggtg gttttcccca
     1321 tcccgtgagg accgtctata taaacccgca gtagcgtggg cattttctgc tccaggcgga
     1381 cttccgtggc ttcttgctgc cggcgagggc gcaacgccgt acgtcggttg ctatggccgc
     1441 gagaacgcgc agcctggtcg aacgcagacg cgtgttgatg gcaggggtac gaagccatgg
     1501 atctgaagat tgggttcctc tttgagagag ttaatagact tgaattgagg aatggaaatc
     1561 gggctgagct cactgcttta tatgtgagaa aaggtgatga gcaggtggta agaggcagat
     1621 ttttgtggtc ggtgctgtgt tttgttgggt ttgagctagg tggggtgagg gaggggcaag
     1681 ctaaatagct accagccctg aggacgtcac agagttttag tagttctttt tctgccatcg
     1741 gacatgcagc tcacgcctag acccaattac accctttgcg ccctcccaca catccggccg
     1801 gttggatcat tgtcagatac ggcagagaaa tcgcaacctc ggccaagggt agccacgatt
     1861 cgaagccgct acttgagacc actcatcgcg actagtagcg gccgctgcag tccggcaaaa
     1921 aagggcaagg tgtcaccacc ctgccctttt tctttaaaac cgaaaagatt acttcgcgtt
     1981 atgcaggctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     2041 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     2101 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     2161 tttttccaca ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     2221 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     2281 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     2341 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     2401 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     2461 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     2521 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     2581 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     2641 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     2701 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     2761 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     2821 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     2881 aaatcaatct aaagtatata tgagtaaact tggtctgaca gctcgaggct tggattctca
     2941 ccaataaaaa acgcccggcg gcaaccgagc gttctgaaca aatccagatg gagttctgag
     3001 gtcattactg gatctatcaa caggagtcca agcgagctcg atatcaaatt acgccccgcc
     3061 ctgccactcg tcgcagtact gttgtaattc attaagcatt ctgccgacat ggaagccatc
     3121 acaaacggca tgatgaacct gaatcgccag cggcatcagc accttgtcgc cttgcgtata
     3181 atatttgccc atggtgaaaa cgggggcgaa gaagttgtcc atattggcca cgtttaaatc
     3241 aaaactggtg aaactcaccc agggattggc tgatacgaaa aacatattct caataaaccc
     3301 tttagggaaa taggccaggt tttcaccgta acacgccaca tcttgcgaat atatgtgtag
     3361 aaactgccgg aaatcgtcgt ggtattcact ccagagtgat gaaaacgttt cagtttgctc
     3421 atggaaaacg gtgtaacaag ggtgaacact atcccatatc accagctcac cgtctttcat
     3481 tgccatacga aattccggat gagcattcat caggcgggca agaatgtgaa taaaggccgg
     3541 ataaaacttg tgcttatttt tctttacggt ctttaaaaag gccgtaatat ccagctgaac
     3601 ggtctggtta taggtacatt gagcaactga ctgaaatgcc tcaaaatgtt ctttacgatg
     3661 ccattgggat atatcaacgg tggtatatcc agtgattttt ttctccattt tagcttcctt
     3721 agctcctgaa aatctcgata actcaaaaaa tacgcccggt agtgatctta tttcattatg
     3781 gtgaaagttg gaacctctta cgtgcccgat caactcgagt gccacctgac gtctaagaaa
     3841 ccattattat catgacatta acctataaaa ataggcgtat cacgaggcag a
//
