LOCUS       sequence-465794-        4554 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Serves as a template plasmid for amplifying Shark2 (Tet-ON) 
            hammerhead aptazyme-3xTy-tagging cassette.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4554)
  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 4554)
  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-Shark2
FEATURES             Location/Qualifiers
     source          1..4554
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      711..1299
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1200..1219
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1569..1967)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     promoter        4316..4379
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4380..4448,4449..4554,1..686)
                     /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 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
       61 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
      121 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
      181 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
      241 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa cattggggat catgtaactc
      301 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
      361 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
      421 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
      481 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
      541 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
      601 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
      661 gtgcctcact gattaagcat tggtaacgta gaaaagatca aaggatcttc ttgagatcct
      721 tttttcctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
      781 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
      841 cggataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
      901 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
      961 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     1021 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     1081 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     1141 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     1201 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     1261 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa accagccgtg cacacgaggt
     1321 acccgcggcg acacggaggg agtggggtgc atggggatgc tgagtcccgc gaagtgtgct
     1381 gggtaattac ggtgcagtca aaactgacaa ataatgcaag tcattgtttg cattttccca
     1441 ctgatatttt taaacaggtt aaatttgctt tgttttcccc cgtgttgttg aacgtttgaa
     1501 tcatttttgc taatccgtgc ttattttgat aaaaaaagac aaatataatg agagtagagg
     1561 tatccgtgtt agccctccca cacataacca gagggcagca attcacgaat cccaactgcc
     1621 gtcggctgtc catcactgtc cttcactatg gctttgatcc caggatgcag atcgagaagc
     1681 acctgtcggc accgtccgca ggggctcaag atgcccctgt tctcatttcc gatcgcgacg
     1741 atacaagtca ggttgccagc tgccgcagca gcagcagtgc ccagcaccac gagttctgca
     1801 caaggtcccc cagtaaaatg atatacattg acaccagtga agatgcggcc gtcgctagag
     1861 agagctgcgc tggcgacgct gtagtcttca gagatgggga tgctgttgat tgtagccgtt
     1921 gctctttcaa tgagggtgga ttcttcttga gacaaaggct tggccataat gcacaaacac
     1981 taaactacac cttaaacacc atcagtgaag aaaaagaaaa aaaaagaaaa acagcttaaa
     2041 tccttaaaaa aaaaaaatca cactaaatta aaacagtcag ttataaacaa aaagaaactg
     2101 aacaaccaaa atcttccaat aacgcaaagt tgtaaaaaat taccacaagc tgcatcatac
     2161 acccaatcaa atcatctaca ttttccttgc aataccccaa atcctcgtac attgattaaa
     2221 cattatttca ctcatccttc cgcatcatta aattgctgct ttttgtttcc caaagatgac
     2281 cataaatata atttcaaaaa aaaaaaagaa aaaaaagtta tatatttttt tgttgttgtc
     2341 cttaaaggga aaagtaaaat tgttttgctc ttgacactgt gtcacttgga tcgtggcagc
     2401 accggtaacg atcatcacgg gacacgcatc gtgtattata agatccctat tcgcaaaaga
     2461 aagacctgaa gtcatcgcca cgctttgagc gcgtttcgtc ctatttggga ctcatcagct
     2521 ggatgtacag gtggtcgtat tcttcacctc tccagatcga aatctggtat gttttgtacg
     2581 aatccaggac gcgcctttgt taagcgtcca ggggatcctg attggtgtgc acttcggcat
     2641 ccagaggatc ctgatttgta tggacttcgg catcaagggg atcttgatta gtatgcactt
     2701 ccttgtacag ttcatccatg cctccagttg aatggcggcc ttccgctcgc tcgtactgct
     2761 caactattgt atagtcttcg ttgtggctag tgatatcgag ctttatgttc acattataag
     2821 ctcccggcaa ttggactggc ttcttagcct tataagttgt tttaacttcc gcgtcgtaat
     2881 ggccgccatc cttaagttta agacgttgct taatttcacc tttgagagcc ccgtcctcag
     2941 ggtacatcct ttcggaagag gcctcccacc ccatcgtttt cttctgcatc accggtccgt
     3001 cggaagggaa atttgtgccc cgcaacttga ccttgtagat gaactccaca tcctgcaagg
     3061 aactatcctg ggtgacggtc acgacaccgc catcctcgaa attcataacg cgttcccatt
     3121 tgaatccttc tgggaagctg agcttcaggt agtcagggat gtctgccggg tgcttgacgt
     3181 acgccttaga gccatacatg aactgaggag acagaatgtc ccaagcgaag ggaagcggcc
     3241 ctcccttcgt gaccttgagt ttcgcagttt gagtcccttc atagggcctg ccttctccct
     3301 ccccttctat ctcgaactca tggccattaa cactgccctc catgtggacc ttgaatcgca
     3361 taaactcctt gatgatggcc atattgtctt cttcaccttt cgacaccatt ctagacaaaa
     3421 aaaaaccccg ccctgtcagg ggcggggttt ttttttagac aacttataga gcaaaaaaaa
     3481 aagaaaagac atcataaaag aagttggaaa agaaaaaagg taaaaacaat ttaacacaac
     3541 aataataagt ggataaaaca acaaagaaaa agtattataa taacaatcat aataatttcg
     3601 ctttcgtgcg tgaaaatgtt tcttagtgaa tccgctgcat attttgaaga aactcgttgg
     3661 gggatccact agaactcttg cacagacttg gcatccacac ggcgctttga ctcccattct
     3721 gaaaaaaaaa agaacacgtt gattcccctt ttgatttccc acgaatgcga agcacaattc
     3781 acgcacagcc aacggcaggg agagacgata aggtaaaata cacacaagtg gtggaatcac
     3841 aaaaggacac gcacagcacg caaataaaca ttgcacacgg catgacattt tgcgacccac
     3901 acacaatgtg tacactacaa ttatatgaaa atccctcccc cattccgcgg gtgcgccgca
     3961 aagggccaaa aacccaaatg atccacttta ttataaacct cactgaccaa tgagccaaaa
     4021 agaaggtcgc gccgcacacg atacggcgat gcgaaacaaa agaccgcaaa aaaaaaacga
     4081 aaaaccaaac atcctgaggg acaccgattc cttccgacgc cacctgggcc accggagaga
     4141 ataccgctcc atgatgtact agacacaaat ggacggttaa aaaagcacaa gttggatcga
     4201 accccggaaa ctccaaaatt tttttagggg tacccaaaaa ttgcaaaaaa ttcctccgaa
     4261 tcatcaaaaa attaaaaatc caacacacca aataagcccc gcaaggaaac tggtgtatgt
     4321 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     4381 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     4441 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     4501 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttc
//
