LOCUS       sequence-251974-        1877 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes c120 sg RNA guide cleaved by ribozyme.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 1877)
  AUTHORS   Fonseca JP, Bonny AR, Kumar GR, Ng AH, Town J, Wu QC, Aslankoohi E, 
            Chen SY, Dods G, Harrigan P, Osimiri LC, Kistler AL, El-Samad H
  TITLE     A Toolkit for Rapid Modular Construction of Biological Circuits in 
            Mammalian Cells
  JOURNAL   ACS Synth Biol. 2019 Nov 15;8(11):2593-2606. doi: 
            10.1021/acssynbio.9b00322. Epub 2019 Nov 5.
  PUBMED    31686495
REFERENCE   2  (bases 1 to 1877)
  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"; doi: 
            "10.1021/acssynbio.9b00322"; journalName: "ACS Synth Biol"; date: 
            "2019-11-15- 15"; volume: "8"; issue: "11"; pages: "2593-2606"
COMMENT     Sequence Label: MTK3_028
FEATURES             Location/Qualifiers
     source          1..1877
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     278..297
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      545..1133
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1034..1053
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     ncRNA           1254..1290
                     /product="self-cleaving hammerhead ribozyme (Ferre-D'Amare 
                     and Scott, 2010)"
                     /label=hammerhead ribozyme
                     /ncRNA_class="hammerhead_ribozyme"
     misc_RNA        1311..1386
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     ncRNA           1391..1458
                     /product="hepatitis delta virus self-cleaving ribozyme 
                     (Ferre-D'Amare and Scott, 2010)"
                     /label=HDV ribozyme
                     /ncRNA_class="ribozyme"
     terminator      complement(1476..1510)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(join(1581..1877,1..363))
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
ORIGIN
        1 cgaatatatg tgtagaaact gccggaaatc gtcgtggtat tcactccaga gcgatgaaaa
       61 cgtttcagtt tgctcatgga aaacggtgta acaagggtga acactatccc atatcaccag
      121 ctcaccgtct ttcattgcca tacgaaattc cggatgagca ttcatcaggc gggcaagaat
      181 gtgaataaag gccggataaa acttgtgctt atttttcttt acggtcttta aaaaggccgt
      241 aatatccagc tgaacggtct ggttataggt acattgagca actgactgaa atgcctcaaa
      301 atgttcttta cgatgccatt gggatatatc aacggtggta tatccagtga tttttttctc
      361 cattttagct tccttagctc ctgaaaatct cgataactca aaaaatacgc ccggtagtga
      421 tcttatttca ttatggtgaa agttggaacc tcttacgtgc ccgatcaatc atgaccaaaa
      481 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
      541 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
      601 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
      661 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
      721 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
      781 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
      841 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
      901 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
      961 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     1021 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     1081 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     1141 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     1201 ctgcgttatc ccctgattct gtggataacc gtagtcggtc tcatatgaac gtactgatga
     1261 gtccgtgagg acgaaacgag taagctcgtc tacgttctct atcactgata gttttagagc
     1321 tagaaatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt
     1381 cggtgctttt ggccggcatg gtcccagcct cctcgctggc gccggctggg caacatgctt
     1441 cggcatggcg aatgggacat cctgagacca gaccaataaa aaacgcccgg cggcaaccga
     1501 gcgttctgaa caaatccaga tggagttctg aggtcattac tggatctatc aacaggagtc
     1561 caagcgagct cgatatcaaa ttacgccccg ccctgccact catcgcagta ctgttgtaat
     1621 tcattaagca ttctgccgac atggaagcca tcacaaacgg catgatgaac ctgaatcgcc
     1681 agcggcatca gcaccttgtc gccttgcgta taatatttgc ccatggtgaa aacgggggcg
     1741 aagaagttgt ccatattggc cacgtttaaa tcaaaactgg tgaaactcac ccagggattg
     1801 gctgaaacga aaaacatatt ctcaataaac cctttaggga aataggccag gttttcaccg
     1861 taacacgcca catcttg
//
