LOCUS       sequence-142819-        5554 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Generates truncated version of MAK10 gene, which is required for 
            dsRNA L-A propagation in S.cerevisiae.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5554)
  TITLE     McCusker_pUG-amdSYM-mak10TR
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5554)
  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            "Unpublished"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: pUG-amdSYM-mak10TR
FEATURES             Location/Qualifiers
     source          1..5554
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        807..825
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     807..824
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    876..909
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     terminator      2969..3166
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     protein_bind    3188..3221
                     /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(3274..3293)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3275..3293)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3380..3397)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3551..4139)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3631..3650)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4310..5101,5102..5170))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4933..4952
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5171..5275)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5343..5361
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5399..5421)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5521..5540
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tatgattcaa tatcattttg actatctttc aaaaaggttt ctaagtggtg acttaaataa
       61 taaacatacc aaaacataga gtatgcttca aagggtttgt aaatatccaa atcaaacccc
      121 tttaaagaga attcgatcat acttgatagt ttcataaaat atgtccacga acagtaaact
      181 tgtgaattga cactttcaaa ctgggcttga agagaatccc atagaataag ttgcctgtta
      241 aaaccttgtc tatatctaca tgtgttttgg gagcagtttt ggtaccattc taaaagcata
      301 ttagagcttt cttggataag ttcattacca atagaggatt caaattcaga tggagtttgt
      361 gctgaaaact ccttgacatg tagcaaataa aattgggtat atgaaaattt acccaaaact
      421 gttctgtcgt ctctaatgaa aaataagggg aataagatcc ttgcaatgac atgcctttgt
      481 tccaacttgt tgaagaattt ggcaaactgg taggtctcca acgcagactc ggctttatct
      541 acaaggagta ttgttttaac atcatttgcc agtgtaataa agccgctgta atccgtcggt
      601 aacttcgtaa tcttcctggg agggaattga ttagacctgt gcttttgtat atatgttgag
      661 aacgcaccct ttggtggaga cagctgtaat tgtggaaata tgccattaac cgagttagcg
      721 ttttcaatta gctcatccca tatggacata ttgtcgttag aacgcggcta caattaatac
      781 ataaccttat gtatcataca catacgattt aggtgacact atagaacgcg gccgccagct
      841 gaagcttcgt acgctgcagg tcgacaaccc ttaatataac ttcgtataat gtatgctata
      901 cgaagttatt aggtctagag atctgtttag cttgcctcgt ccccgccggg tcacccggcc
      961 agcgacatgg aggcccagaa taccctcctt gacagtcttg acgtgcgcag ctcaggggca
     1021 tgatgtgact gtcgcccgta catttagccc atacatcccc atgtataatc atttgcatcc
     1081 atacattttg atggccgcac ggcgcgaagc aaaaattacg gctcctcgct gcagacctgc
     1141 gagcagggaa acgctcccct cacagacgcg ttgaattgtc cccacgccgc gcccctgtag
     1201 agaaatataa aaggttagga tttgccactg aggttcttct ttcatatact tccttttaaa
     1261 atcttgctag gatacagttc tcacatcaca tccgaacata aacaaggatc catgccacaa
     1321 tcttgggaag aattggctgc tgacaagaga gctagattgg ctaagactat cccagacgaa
     1381 tggaaggttc aaactttgcc agctgaagac tctgttatcg acttcccaaa gaagtctggt
     1441 atcttgtctg aagctgaatt gaagatcact gaagcttctg ctgctgactt ggtttctaag
     1501 ttggctgctg gtgaattgac ttctgttgaa gttactttgg ctttctgtaa gagagctgct
     1561 atcgctcaac aattgactaa ctgtgctcac gaattcttcc cagacgctgc tttggctcaa
     1621 gctagagaat tggacgaata ctacgctaag cacaagagac cagttggtcc attgcacggt
     1681 ttgccaatct ctttgaagga ccaattgaga gttaagggtt acgaaacttc tatgggttac
     1741 atctcttggt tgaacaagta cgacgaaggt gactctgttt tgactactat gttgagaaag
     1801 gctggtgctg ttttctacgt taagacttct gttccacaaa ctttgatggt ttgtgaaact
     1861 gttaacaaca tcatcggtag aactgttaac ccaagaaaca agaactggtc ttgtggtggt
     1921 tcttctggtg gtgaaggtgc tatcgttggt atcagaggtg gtgttatcgg tgttggtact
     1981 gacatcggtg gttctatcag agttccagct gctttcaact tcttgtacgg tttgagacca
     2041 tctcacggta gattgccata cgctaagatg gctaactcta tggaaggtca agaaactgta
     2101 cactctgttg ttggtccaat cactcactct gttgaagact tgagattgtt cactaagtct
     2161 gttttgggtc aagaaccatg gaagtacgac tctaaggtta tcccaatgcc atggagacaa
     2221 tctgaatctg acatcatcgc ttctaagatc aagaacggtg gtttgaacat cggttactac
     2281 aacttcgacg gtaacgtttt gccacaccca ccaatcttga gaggtgttga aactactgtt
     2341 gctgctttgg ctaaggctgg tcacactgtt actccatgga ctccatacaa gcacgacttc
     2401 ggtcacgact tgatctctca catctacgct gctgacggtt ctgctgacgt tatgagagac
     2461 atctctgctt ctggtgaacc agctatccca aacatcaagg acttgttgaa cccaaacatc
     2521 aaggctgtta acatgaacga attgtgggac actcacttgc aaaagtggaa ctaccaaatg
     2581 gaatacttgg aaaagtggag agaagctgaa gaaaaggctg gtaaggaatt ggacgctatc
     2641 atcgctccaa tcactccaac tgctgctgtt agacacgacc aattcagata ctacggttac
     2701 gcttctgtta tcaacttgtt ggacttcact tctgttgttg ttccagttac tttcgctgac
     2761 aagaacatcg acaagaagaa cgaatctttc aaggctgttt ctgaattgga cgctttggtt
     2821 caagaagaat acgacccaga agcttaccac ggtgctccag ttgctgttca agttatcggt
     2881 agaagattgt ctgaagaaag aactttggct atcgctgaag aagttggtaa gttgttgggt
     2941 aacgttgtta ctccataact cgagcagtac tgacaataaa aagattcttg ttttcaagaa
     3001 cttgtcattt gtatagtttt tttatattgt agttgttcta ttttaatcaa atgttagcgt
     3061 gatttatatt ttttttcgcc tcgacatcat ctgcccagat gcgaagttaa gtgcgcagaa
     3121 agtaatatca tgcgtcaatc gtatgtgaat gctggtcgct atactggagc tctcgagaac
     3181 ccttaatata acttcgtata atgtatgcta tacgaagtta ttaggtgata tcagatccac
     3241 tagtggccta tgcggccgcg gatctgccgg tctccctata gtgagtcgta ttaatttcga
     3301 taagccaggt taacctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     3361 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     3421 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     3481 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     3541 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     3601 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     3661 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     3721 cttcgggaag cgtggcgctt tctcaatgct cacgctgtag gtatctcagt tcggtgtagg
     3781 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     3841 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     3901 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     3961 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     4021 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     4081 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     4141 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     4201 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     4261 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     4321 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     4381 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     4441 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     4501 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     4561 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     4621 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     4681 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     4741 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     4801 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     4861 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     4921 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     4981 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     5041 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     5101 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     5161 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     5221 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     5281 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
     5341 aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc
     5401 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca
     5461 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg
     5521 cggcatcaga gcagattgta ctgagagtgc acca
//
