LOCUS       sequence-465795-        4595 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Serves as a template plasmid for amplifying Shark1 (Tet-OFF) 
            hammerhead aptazyme-3xTy-tagging cassette.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4595)
  AUTHORS   Wiedeman J, Harrison R, Etheridge RD
  TITLE     A limitation lifted: A conditional knockdown system reveals 
            essential roles for Polo-like kinase and Aurora kinase 1 in 
            Trypanosoma cruzi cell division.
  JOURNAL   Proc Natl Acad Sci U S A. 2025 Feb 25;122(8):e2416009122. doi: 
            10.1073/pnas.2416009122. Epub 2025 Feb 18.
  PUBMED    40106484
REFERENCE   2  (bases 1 to 4595)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2025-02-25"; volume: "122(8):e2416009122. 
            doi"; pages: " 10.1073/pnas.2416009122. Epub 2025 Feb 18"
COMMENT     Sequence Label: pTMiniTrex-mCherry-Shark1
FEATURES             Location/Qualifiers
     source          1..4595
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     550..569
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(788..851)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3241..3639
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     rep_origin      complement(3909..4497)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3989..4008)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4522..4595,1..718,719..787))
                     /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"
ORIGIN
        1 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
       61 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
      121 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
      181 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
      241 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
      301 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
      361 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
      421 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
      481 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
      541 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
      601 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
      661 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
      721 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
      781 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
      841 gcggatacat acaccagttt ccttgcgggg cttatttggt gtgttggatt tttaattttt
      901 tgatgattcg gaggaatttt ttgcaatttt tgggtacccc taaaaaaatt ttggagtttc
      961 cggggttcga tccaacttgt gcttttttaa ccgtccattt gtgtctagta catcatggag
     1021 cggtattctc tccggtggcc caggtggcgt cggaaggaat cggtgtccct caggatgttt
     1081 ggtttttcgt tttttttttt gcggtctttt gtttcgcatc gccgtatcgt gtgcggcgcg
     1141 accttctttt tggctcattg gtcagtgagg tttataataa agtggatcat ttgggttttt
     1201 ggccctttgc ggcgcacccg cggaatgggg gagggatttt catataattg tagtgtacac
     1261 attgtgtgtg ggtcgcaaaa tgtcatgccg tgtgcaatgt ttatttgcgt gctgtgcgtg
     1321 tccttttgtg attccaccac ttgtgtgtat tttaccttat cgtctctccc tgccgttggc
     1381 tgtgcgtgaa ttgtgcttcg cattcgtggg aaatcaaaag gggaatcaac gtgttctttt
     1441 tttttcagaa ggggagtcaa agcgccgtgt ggatgccaag tctgtgcaag agttctagtg
     1501 gatcccccaa cgagtttctt caaaatatgc agcggattca ctaagaaaca ttttcacgca
     1561 cgaaagcgaa attattatga ttgttattat aatacttttt ctttgttgtt ttatccactt
     1621 attattgttg tgttaaattg tttttacctt ttttcttttc caacttcttt tatgatgtct
     1681 tttctttttt ttttgctcta taagttgtct aaaaaaaaac cccgcccctg acagggcggg
     1741 gttttttttt gtctagaatg gtgtcgaaag gtgaagaaga caatatggcc atcatcaagg
     1801 agtttatgcg attcaaggtc cacatggagg gcagtgttaa tggccatgag ttcgagatag
     1861 aaggggaggg agaaggcagg ccctatgaag ggactcaaac tgcgaaactc aaggtcacga
     1921 agggagggcc gcttcccttc gcttgggaca ttctgtctcc tcagttcatg tatggctcta
     1981 aggcgtacgt caagcacccg gcagacatcc ctgactacct gaagctcagc ttcccagaag
     2041 gattcaaatg ggaacgcgtt atgaatttcg aggatggcgg tgtcgtgacc gtcacccagg
     2101 atagttcctt gcaggatggg gagttcatct acaaggtcaa gttgcggggc acaaatttcc
     2161 cttccgacgg accggtgatg cagaagaaaa cgatggggtg ggaggcctct tccgaaagga
     2221 tgtaccctga ggacggggct ctcaaaggtg aaattaagca acgtcttaaa cttaaggatg
     2281 gcggccatta cgacgcggaa gttaaaacaa cttataaggc taagaagcca gtccaattgc
     2341 cgggagctta taatgtgaac ataaagctcg atatcactag ccacaacgaa gactatacaa
     2401 tagttgagca gtacgagcga gcggaaggcc gccattcaac tggaggcatg gatgaactgt
     2461 acaaggaagt gcatactaat caagatcccc ttgatgccga agtccataca aatcaggatc
     2521 ctctggatgc cgaagtgcac accaatcagg atcccctgga cgcttaacaa aaaacaaaca
     2581 aactgaggtg caggtacatc cagctgacga gtcccaaata ggacgaaagt taaaacatac
     2641 cagatttcga tctggagagg tgaagaatac gaccacctaa ttcctggatt caactgctat
     2701 ccacaaaaag aaaaataaaa acaaagcgtg gcgatgactt caggtctttc ttttgcgaat
     2761 agggatctta taatacacga tgcgtgtccc gtgatgatcg ttaccggtgc tgccacgatc
     2821 caagtgacac agtgtcaaga gcaaaacaat tttacttttc cctttaagga caacaacaaa
     2881 aaaatatata actttttttt cttttttttt tttgaaatta tatttatggt catctttggg
     2941 aaacaaaaag cagcaattta atgatgcgga aggatgagtg aaataatgtt taatcaatgt
     3001 acgaggattt ggggtattgc aaggaaaatg tagatgattt gattgggtgt atgatgcagc
     3061 ttgtggtaat tttttacaac tttgcgttat tggaagattt tggttgttca gtttcttttt
     3121 gtttataact gactgtttta atttagtgtg attttttttt ttaaggattt aagctgtttt
     3181 tctttttttt tctttttctt cactgatggt gtttaaggtg tagtttagtg tttgtgcatt
     3241 atggccaagc ctttgtctca agaagaatcc accctcattg aaagagcaac ggctacaatc
     3301 aacagcatcc ccatctctga agactacagc gtcgccagcg cagctctctc tagcgacggc
     3361 cgcatcttca ctggtgtcaa tgtatatcat tttactgggg gaccttgtgc agaactcgtg
     3421 gtgctgggca ctgctgctgc tgcggcagct ggcaacctga cttgtatcgt cgcgatcgga
     3481 aatgagaaca ggggcatctt gagcccctgc ggacggtgcc gacaggtgct tctcgatctg
     3541 catcctggga tcaaagccat agtgaaggac agtgatggac agccgacggc agttgggatt
     3601 cgtgaattgc tgccctctgg ttatgtgtgg gagggctaac acggatacct ctactctcat
     3661 tatatttgtc tttttttatc aaaataagca cggattagca aaaatgattc aaacgttcaa
     3721 caacacgggg gaaaacaaag caaatttaac ctgtttaaaa atatcagtgg gaaaatgcaa
     3781 acaatgactt gcattatttg tcagttttga ctgcaccgta attacccagc acacttcgcg
     3841 ggactcagca tccccatgca ccccactccc tccgtgtcgc cgcgggtacc tcgtgtgcac
     3901 ggctggtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     3961 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     4021 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     4081 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     4141 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     4201 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     4261 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     4321 tggtggccta actacggcta cactagaaga acagtatttg gtatccgcgc tctgctgaag
     4381 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     4441 agcggtggtt tttttgtttg caagcagcag attacgcgca ggaaaaaagg atctcaagaa
     4501 gatcctttga tcttttctac gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     4561 ctgtctattt cgttcatcca tagttgcctg actcc
//
