LOCUS       4_U6_ISCro4_(WT)        2579 bp    DNA     circular SYN 05-JAN-2026
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2579)
  AUTHORS   Weatherall Institute
  TITLE     Direct Submission
  JOURNAL   Exported Jan 5, 2026 from SnapGene 8.2.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..2579
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(26..42)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(214..802)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    complement(919..943)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(join(1043..1834,1835..1903))
                     /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"
     promoter        complement(1904..2008)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2122..2138
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2191..2439
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_feature    2440
                     /label=+G
     misc_feature    2441..2542
                     /label=ISCro4 TBL (target binding loop)
     misc_feature    2543..2548
                     /label=polymerase III terminator
ORIGIN
        1 tgcgatggtg gatgcatgtg tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
       61 tcagagggca caatcctatt ccgcgctatc cgacaatctc caagacatta ggtggagttc
      121 agttcggcgt atggcatatg tcgctggaaa gaacatgtga gcaaaaggcc agcaaaaggc
      181 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
      241 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
      301 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
      361 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
      421 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
      481 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
      541 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
      601 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
      661 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
      721 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
      781 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctct
      841 attcaacaaa gccgccgtcc cgtcaagtca gcgtaaatgg gtagggggct tcaaatcgtc
      901 ctcgtgatac caattcggag cctgcttttt tgtacaaact tgttgataat ggcaattcaa
      961 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     1021 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     1081 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     1141 gggagggctt accatctggc cccagtgctg caatgatacc gcgagagcca cgctcaccgg
     1201 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     1261 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     1321 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     1381 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     1441 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     1501 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     1561 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     1621 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     1681 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     1741 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     1801 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     1861 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     1921 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     1981 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca gatacctgaa
     2041 acaaaaccca tcgtacggcc aaggaagtct ccaataactg tgatccacca caagcgccag
     2101 ggttttccca gtcacgacgt tgtaaaacga cggccagtca tgcataatcc gcacgcatct
     2161 ggaataagga agtgccattc cgcctgacct gagggcctat ttcccatgat tccttcatat
     2221 ttgcatatac gatacaaggc tgttagagag ataattggaa ttaatttgac tgtaaacaca
     2281 aagatattag tacaaaatac gtgacgtaga aagtaataat ttcttgggta gtttgcagtt
     2341 ttaaaattat gttttaaaat ggactatcat atgcttaccg taacttgaaa gtatttcgat
     2401 ttcttggctt tatatatctt gtggaaagga cgaaacaccg agtgcaggga gaaccggcca
     2461 gttctctctg ccatgcggtc cgcatgccgt atcaggcctc aggctaataa cctgtgacgt
     2521 agatttatgc agcggaccgc cgttttttag gctaggtgga ggctcagtga tgataagtc
//
