LOCUS       sequence-431514-        5240 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fragmid fragment: (Cas protein) Cas13d.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5240)
  AUTHORS   McGee AV, Liu YV, Griffith AL, Szegletes ZM, Wen B, Kraus C, Miller 
            NW, Steger RJ, Escude Velasco B, Bosch JA, Zirin JD, Viswanatha R, 
            Sontheimer EJ, Goodale A, Greene MA, Green TM, Doench JG
  TITLE     Modular vector assembly enables rapid assessment of emerging CRISPR 
            technologies.
  JOURNAL   Cell Genom. 2024 Mar 13;4(3):100519. doi: 
            10.1016/j.xgen.2024.100519.
  PUBMED    38484704
REFERENCE   2  (bases 1 to 5240)
  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: "Cell 
            Genom."; date: "2024-03-13"; volume: "4(3):100519. doi"; pages: " 
            10.1016/j.xgen.2024.100519"
COMMENT     Sequence Label: AA345
FEATURES             Location/Qualifiers
     source          1..5240
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(3150..3166)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3150..3166)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3163..3185)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3174..3190
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3198..3204,3205..3222,3223..3228))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3243..3264
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3381..3398)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3552..4140)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4318..5127)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     5041..5060
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5119..5138
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagagaatt cgagctcggt acctcgcgaa
      421 tacatctaga tgaagactgg cagcgccacc aagaaacaga agtccaagaa aaccgtgagc
      481 aagaccagcg gcctgaagga agccctgagt gtgcagggca ccgtcatcat gacctccttc
      541 ggcaaaggaa acatggccaa cctgagctac aaaattcctt cttctcagaa acctcagaac
      601 ctgaatagca gcgctggcct taagaacgtg gaagtgtccg gaaagaagat caagttccag
      661 ggcagacacc ctaagattgc caccaccgac aaccctctgt tcaagccaca acctggcatg
      721 gacctgctgt gcctgaaaga caaactggaa atgcactact tcggcaagac cttcgacgac
      781 aacatccaca tacagctcat ctaccagatc ctggacatcg agaagatcct ggccgtgcac
      841 gtgaacaaca tcgtgttcac cctggataat gttctgcacc cccagaagga agaactgaca
      901 gaggacttta tcggagccgg cggatggcgg atcaaccttg actaccagac actgagaggc
      961 cagaccaata aatacgacag attcaagaac tacatcaaaa gaaaggaact gctgtatttc
     1021 ggcgaggcct tttaccacga gaacgagcgg agatatgaag aggacatctt cgccatcctg
     1081 acactgctgt cggcacttcg gcagttctgc ttccacagcg atctgtccag cgacgagtct
     1141 gaccacgtga actctttctg gctgtaccag ctggaggacc agctgagcga cgagtttaaa
     1201 gaaaccctga gcatcctgtg ggaggaggtg accgagagga tcgactccga gttcctgaaa
     1261 accaacaccg tgaatctgca catcctgtgc cacgtgttcc ccaaggaatc taaagaaacc
     1321 atcgtgcggg cctactacga gttcctgatt aagaagagct ttaagaacat gggcttcagc
     1381 atcaaaaaac tgcgggaaat catgctggaa cagagcgacc tgaagtcttt taaagaagat
     1441 aagtacaatt ccgtgcgggc caagctgtac aagctgtttg atttcatcat cacctactac
     1501 tatgatcacc acgcctttga gaaggaagcc ctggttagca gcctgagatc ttctctgacg
     1561 gaggaaaaca aagaggaaat ctacatcaag acagccagaa cactcgccag cgccctgggc
     1621 gccgatttca agaaggctgc tgctgatgtt aacgccaaga acatcagaga ctaccagaaa
     1681 aaggccaacg actaccggat cagcttcgag gacatcaaga ttggaaatac cggcatcggc
     1741 tactttagcg agctgatcta catgctgaca ctgctgctgg acggcaagga aatcaacgac
     1801 ctgctgacta cactgatcaa caagttcgat aacatcatct cttttatcga catcctgaag
     1861 aagctgaacc tggaattcaa gttcaagcct gagtacgccg atttcttcaa catgacaaac
     1921 tgcagataca ccctggaaga actgagagtg atcaacagca tcgccaggat gcagaagcct
     1981 agcgccgacg ccaggaagat catgtacaga gatgctttaa gaatcctggg catggacaat
     2041 agacctgatg aggagattga tagagagctg gagagaacca tgcctgtggg agccgatggc
     2101 aagtttatca agggaaagca aggttttcgg aatttcatcg cctctaatgt catcgagagc
     2161 agccgtttcc attacctggt gcggtacaac aacccccaca agaccagaac actggtgaag
     2221 aatcccaacg tggtgaagtt cgtgctggag ggcatcccag agacccagat caagcggtac
     2281 ttcgatgtgt gcaagggcca ggagataccc cccacaagcg acaagagcgc ccaaatcgac
     2341 gtgctggcca gaatcatcag cagcgtggac tacaagatct tcgaggacgt gccccaaagt
     2401 gccaagatca acaaggacga tcctagcaga aacttcagcg acgcactgaa gaagcaaaga
     2461 taccaggcca tcgtgtccct gtacctgaca gtgatgtacc tgatcaccaa gaacctggtg
     2521 tacgtgaact cccggtacgt gatcgccttc cactgtctgg aacgcgacgc cttcctgcac
     2581 ggcgtgacac tccccaagat gaacaagaag atcgtgtaca gccagctgac cacccacctg
     2641 ctcaccgaca agaactatac aacctacggc cacctgaaga accagaaagg ccaccggaag
     2701 tggtatgtgc tggtgaagaa caacctgcag aattctgata tcacagccgt gagctccttc
     2761 agaaacatcg tcgcccacat cagcgtggtc agaaatagca acgagtacat cagcggcatc
     2821 ggcgaactgc attcttattt cgagctgtac cactacctgg tccagagcat gatcgctaag
     2881 aacaattggt acgatacaag ccaccagcct aagaccgctg agtacctgaa caacctgaag
     2941 aagcaccata cctactgtaa agacttcgtg aaggcttact gcatcccttt cggctatgtg
     3001 gtgccaagat acaagaacct gaccatcaac gaactattcg acagaaacaa ccctaaccct
     3061 gagcctaaag aggaagtggg gtccaagtct tcatcggatc ccgggcccgt cgactgcaga
     3121 ggcctgcatg caagcttggt gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     3181 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     3241 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     3301 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     3361 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     3421 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     3481 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     3541 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     3601 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     3661 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     3721 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     3781 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     3841 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     3901 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     3961 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     4021 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     4081 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     4141 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     4201 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     4261 tgaagtttta aatcaagccc aatctgaata atgttacaac caattaacca attctgatta
     4321 gaaaaactca tcgagcatca aatgaaactg caatttattc atatcaggat tatcaatacc
     4381 atatttttga aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc agttccatag
     4441 gatggcaaga tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa tacaacctat
     4501 taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag tgacgactga
     4561 atccggtgag aatggcaaaa gtttatgcat ttctttccag acttgttcaa caggccagcc
     4621 attacgctcg tcatcaaaat cactcgcatc aaccaaaccg ttattcattc gtgattgcgc
     4681 ctgagcgaga cgaaatacgc gatcgctgtt aaaaggacaa ttacaaacag gaatcgaatg
     4741 caaccggcgc aggaacactg ccagcgcatc aacaatattt tcacctgaat caggatattc
     4801 ttctaatacc tggaatgctg tttttccggg gatcgcagtg gtgagtaacc atgcatcatc
     4861 aggagtacgg ataaaatgct tgatggtcgg aagaggcata aattccgtca gccagtttag
     4921 tctgaccatc tcatctgtaa catcattggc aacgctacct ttgccatgtt tcagaaacaa
     4981 ctctggcgca tcgggcttcc catacaagcg atagattgtc gcacctgatt gcccgacatt
     5041 atcgcgagcc catttatacc catataaatc agcatccatg ttggaattta atcgcggcct
     5101 cgacgtttcc cgttgaatat ggctcataac accccttgta ttactgttta tgtaagcaga
     5161 cagttttatt gttcatgatg atatattttt atcttgtgca atgtaacatc agagattttg
     5221 agacacgggc cagagctgca
//
