LOCUS       sequence-465793-        4574 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Serves as a template plasmid for amplifying Shark3 (Theo-OFF) 
            hammerhead aptazyme-3xTy-tagging cassette.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4574)
  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 4574)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-Shark3
FEATURES             Location/Qualifiers
     source          1..4574
                     /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             3220..3618
                     /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(3888..4476)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3968..3987)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4501..4574,1..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
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNIGDHVTRLDRW
                     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 tgatccccaa 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 tttttttttg cggtcttttg tttcgcatcg ccgtatcgtg tgcggcgcga
     1141 ccttcttttt ggctcattgg tcagtgaggt ttataataaa gtggatcatt tgggtttttg
     1201 gccctttgcg gcgcacccgc ggaatggggg agggattttc atataattgt agtgtacaca
     1261 ttgtgtgtgg gtcgcaaaat gtcatgccgt gtgcaatgtt tatttgcgtg ctgtgcgtgt
     1321 ccttttgtga ttccaccact tgtgtgtatt ttaccttatc gtctctccct gccgttggct
     1381 gtgcgtgaat tgtgcttcgc attcgtggga aatcaaaagg ggaatcaacg tgttcttttt
     1441 tttttcagaa tgggagtcaa 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 gcaggatgtg 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 ggacgaaaga aataccagcc
     2641 gaaaggccct tggcagtttt cctggattca actgctatcc acaaaaagaa aaataaaaac
     2701 aaagcgtggc gatgacttca ggtctttctt ttgcgaatag ggatcttata atacacgatg
     2761 cgtgtcccgt gatgatcgtt accggtgctg ccacgatcca agtgacacag tgtcaagagc
     2821 aaaacaattt tacttttccc tttaaggaca acaacaaaaa aatatataac ttttttttct
     2881 tttttttttt tgaaattata tttatggtca tctttgggaa acaaaaagca gcaatttaat
     2941 gatgcggaag gatgagtgaa ataatgttta atcaatgtac gaggatttgg ggtattgcaa
     3001 ggaaaatgta gatgatttga ttgggtgtat gatgcagctt gtggtaattt tttacaactt
     3061 tgcgttattg gaagattttg gttgttcagt ttctttttgt ttataactga ctgttttaat
     3121 ttagtgtgat tttttttttt taaggattta agctgttttt cttttttttt ctttttcttc
     3181 actgatggtg tttaaggtgt agtttagtgt ttgtgcatta tggccaagcc tttgtctcaa
     3241 gaagaatcca ccctcattga aagagcaacg gctacaatca acagcatccc catctctgaa
     3301 gactacagcg tcgccagcgc agctctctct agcgacggcc gcatcttcac tggtgtcaat
     3361 gtatatcatt ttactggggg accttgtgca gaactcgtgg tgctgggcac tgctgctgct
     3421 gcggcagctg gcaacctgac ttgtatcgtc gcgatcggaa atgagaacag gggcatcttg
     3481 agcccctgcg gacggtgccg acaggtgctt ctcgatctgc atcctgggat caaagccata
     3541 gtgaaggaca gtgatggaca gccgacggca gttgggattc gtgaattgct gccctctggt
     3601 tatgtgtggg agggctaaca cggatacctc tactctcatt atatttgtct ttttttatca
     3661 aaataagcac ggattagcaa aaatgattca aacgttcaac aacacggggg aaaacaaagc
     3721 aaatttaacc tgtttaaaaa tatcagtggg aaaatgcaaa caatgacttg cattatttgt
     3781 cagttttgac tgcaccgtaa ttacccagca cacttcgcgg gactcagcat ccccatgcac
     3841 cccactccct ccgtgtcgcc gcgggtacct cgtgtgcacg gctggttttt ccataggctc
     3901 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     3961 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     4021 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     4081 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     4141 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     4201 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     4261 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     4321 actagaagaa cagtatttgg tatccgcgct ctgctgaagc cagttacctt cggaaaaaga
     4381 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     4441 aagcagcaga ttacgcgcag gaaaaaagga tctcaagaag atcctttgat cttttctacg
     4501 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     4561 agttgcctga ctcc
//
