LOCUS       sequence-431489-        7406 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fragmid fragment: (Cas protein) Cas7-11.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7406)
  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 7406)
  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: AA226
FEATURES             Location/Qualifiers
     source          1..7406
                     /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(5316..5332)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5316..5332)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5329..5351)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5340..5356
                     /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(5364..5370,5371..5388,5389..5394))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5409..5430
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5547..5564)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5718..6306)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5798..5817)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6484..7293)
                     /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     7207..7226
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     7285..7304
                     /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 cagcacgact actatgaaga tttcaattga attcctcgag
      481 ccgtttcgga tgaccaaatg gcaggaaagc actaggcgaa acaagaacaa caaggaattt
      541 gttcgcggcc aagcttttgc aagatggcat agaaataaga aggataacac aaaagggcgg
      601 ccttacataa ctgggacatt gctgcggtcc gcagtaatta gaagcgcgga aaacctgctg
      661 acattgagtg atgggaaaat tagcgaaaag acatgttgtc ctgggaaatt tgatacggaa
      721 gataaagaca ggttgctgca attgaggcag cggtccaccc ttcgttggac cgataagaac
      781 ccctgtccag ataacgctga aacttactgc ccgttttgcg aacttctcgg gagatccggt
      841 aacgatggaa agaaggctga gaagaaggat tggcgctttc gtattcattt tggcaatctt
      901 tcccttccag ggaagcccga tttcgacggg ccaaaagcta taggcagcca acgggtactt
      961 aacagggtcg atttcaaatc cggtaaggcc catgacttct ttaaggctta cgaagtcgac
     1021 catactaggt ttccccgctt cgaaggggag attaccatag ataataaagt cagcgctgaa
     1081 gccaggaaac tgctctgtga ttctctcaag tttacggatc ggctgtgtgg agctctgtgc
     1141 gtaatcaggt tcgatgaata tacaccagca gccgatagtg ggaaacaaac cgaaaatgtc
     1201 caagcagaac cgaatgctaa tctcgctgaa aagaccgcag agcaaattat tagcattctg
     1261 gacgataata agaaaaccga atatacccgc ctgcttgctg atgctattcg ttcactccgc
     1321 cgctctagta aacttgtcgc tggcttgcca aaggatcatg acgggaagga tgaccattac
     1381 ctctgggata tcggcaagaa gaagaaagac gaaaactctg tcactattag gcaaatcctt
     1441 acgacctcag cagacaccaa ggaactcaag aatgccggaa aatggagaga attctgcgag
     1501 aagctgggtg aagcgctgta cctcaagagt aaagacatga gtggcggcct gaaaattact
     1561 cggagaatac tgggcgatgc tgaattccat ggaaagcccg atcggcttga aaagagccgg
     1621 tctgtgtcaa tcggatctgt gttgaaagaa actgtggtat gcggcgaact ggtcgctaaa
     1681 acgcccttct tcttcggagc gatagacgaa gatgcaaaac aaaccgacct gcaagtactc
     1741 ctcactcccg ataacaagta tagactgcca agaagcgccg tgcgaggtat actccgtcga
     1801 gatcttcaaa cctattttga tagcccatgc aatgctgagt tgggtggacg gccatgcatg
     1861 tgtaagacgt gtaggattat gagagggatc acggtgatgg atgcgcgcag tgagtacaat
     1921 gccccgccag aaataaggca tcgtacccgc attaatccct tcacaggcac ggttgcggaa
     1981 ggtgccctgt ttaatatgga agtagccccc gaggggattg tctttccatt tcaactccgg
     2041 taccggggct ctgaagatgg gctgcccgat gcactgaaaa cggtgttgaa atggtgggct
     2101 gaggggcagg cattcatgag tggcgctgcc tcaaccggga agggccgatt ccggatggaa
     2161 aatgctaaat atgaaacgct ggatctgagc gacgagaatc aaaggaatga ctatcttaag
     2221 aattggggat ggcgtgacga gaaggggctc gaggaactga agaaacgact gaactcaggt
     2281 ctgccagagc ccggtaatta tagggatcca aaatggcacg agattaacgt ttccattgag
     2341 atggcaagcc cttttattaa tggcgaccca atccgcgcag ccgtggacaa acgtggtaca
     2401 gatgtggtta ccttcgttaa gtataaagct gaaggggaag aggcgaaacc cgtatgtgca
     2461 tacaaggccg aatcttttag aggggtgatc agaagtgccg tggcacgcat tcatatggaa
     2521 gatggcgtcc ctttgactga gttgactcac agtgactgtg aatgtctcct gtgccaaatc
     2581 tttggaagtg agtatgaagc cggcaaaata aggtttgaag atctcgtatt cgaaagtgac
     2641 ccggaacctg tgaccttcga tcatgtggcc atcgatagat tcactggcgg tgcagctgat
     2701 aagaagaaat tcgatgattc ccctctgccc ggtagccctg caagaccgct catgttgaaa
     2761 ggctccttct ggatccgcag ggacgttctc gaggacgaag agtactgtaa ggcactcggt
     2821 aaggctcttg cagatgtgaa taatggcctt tatcccctcg gtggaaagag cgccatcggc
     2881 tacggacagg tcaagagtct gggtataaag ggagatgata agaggatttc tcgcctcatg
     2941 aatcctgcct ttgatgagac agatgtagcc gttccagaaa agcccaaaac tgatgccgag
     3001 gttcgcatcg aggcagagaa agtatattac ccacactatt tcgtcgaacc ccataagaag
     3061 gtggaacgcg aggagaaacc ctgtggtcat caaaagttcc acgaggggcg actgacaggt
     3121 aaaattcggt gtaagctcat taccaagaca cccctcatcg tcccagatac tagtaatgac
     3181 gatttcttca gacctgcgga taaagaagct cggaaggaaa aggacgaata tcataaatca
     3241 tatgctttct tcagacttca taaacaaatc atgattcccg ggagcgaatt gagaggaatg
     3301 gtgagtagtg tctacgaaac tgtgacaaat tcttgcttca ggatatttga tgagactaaa
     3361 cggttgtcat ggcggatgga tgctgatcac cagaatgtgc tgcaagactt tctcccaggt
     3421 cgagtgaccg ccgatgggaa acatatacaa aagttttccg aaactgcaag ggtgcctttc
     3481 tatgacaaaa cgcagaaaca ttttgacatt cttgatgagc aagaaattgc tggtgagaaa
     3541 cctgttcgga tgtgggtcaa gcgttttatt aaacgactga gcctcgttga tcctgctaaa
     3601 cacccccaga agaaacaaga caataaatgg aaaagacgca aagaaggcat cgccacattt
     3661 atagagcaaa agaatggctc ttattatttt aacgtggtca ccaataatgg ctgcacttct
     3721 ttccacttgt ggcataaacc tgacaatttt gaccaggaga aactcgaagg catacagaat
     3781 ggtgagaaac tggattgctg ggtaagagat agtagatacc aaaaggcctt tcaagagata
     3841 cccgagaacg acccagacgg atgggagtgt aaagagggct accttcatgt cgtcggcccc
     3901 agcaaagtag agtttagtga caagaaaggg gatgtgatca ataactttca aggaacactc
     3961 ccatcagttc ccaacgactg gaaaacaatt aggacgaacg attttaagaa taggaaaagg
     4021 aagaatgaac ctgtgttttg ttgtgaggat gataagggca attactatac catggctaaa
     4081 tattgcgaaa ccttcttctt cgatctgaag gagaacgagg aatacgagat ccccgagaaa
     4141 gcaagaatca aatacaaaga actgctcagg gtctataaca acaatcctca agcagtgccg
     4201 gagagcgtat ttcagtctag agttgcccgg gaaaacgtgg aaaagctgaa gtccggagat
     4261 cttgtgtatt tcaaacataa tgaaaagtac gtagaggaca tcgtcccagt gcggatttcc
     4321 cgaactgtag acgataggat gatcggcaaa cgtatgagcg ccgatctgcg gccgtgccat
     4381 ggagattggg tggaagatgg tgatctcagt gccttgaatg catatcccga gaaaagactc
     4441 ctcttgcgcc accccaaagg actctgccct gcttgccggc tctttggaac cggatcttac
     4501 aagggcagag tcaggtttgg attcgcgtca ctcgaaaacg atccggagtg gctgatccca
     4561 ggcaagaatc ccggcgatcc gtttcacggc gggccggtga tgctctcatt gttggaacgg
     4621 cctcgcccga cttggagtat accgggatcc gacaataagt ttaaagtgcc tggcagaaag
     4681 ttttacgtcc accaccacgc ctggaaaacc attaaggacg ggaaccatcc cacaacaggc
     4741 aaagctattg aacaaagccc taataaccgc actgtagaag ctctcgccgg cgggaattcc
     4801 tttagcttcg aaattgcctt tgagaacctg aaagaatggg agctgggttt gctcatccac
     4861 agcctgcaac tcgaaaaggg tctggcgcat aaacttggaa tggcaaagtc tatgggattt
     4921 ggttcagttg aaattgacgt cgaatcagtg cgcctgagaa aagattggaa gcaatggcgg
     4981 aatggcaatt ccgaaattcc caactggttg ggaaaaggat ttgctaaact gaaggaatgg
     5041 ttccgggacg agctcgattt tatagaaaat cttaagaaac ttctttggtt tcctgagggc
     5101 gaccaagcac cccgggtttg ctaccccatg ctgcgaaaga aggacgatcc taatgggaat
     5161 agcggttacg aagaactcaa agacggggaa ttcaagaaag aagatcggca gaagaagctg
     5221 accacgccgt ggacaccgtg ggcagggtcc aagtcttcat cggatcccgg gcccgtcgac
     5281 tgcagaggcc tgcatgcaag cttggtgtaa tcatggtcat agctgtttcc tgtgtgaaat
     5341 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     5401 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     5461 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     5521 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     5581 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     5641 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     5701 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     5761 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     5821 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     5881 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     5941 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     6001 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     6061 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     6121 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     6181 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     6241 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     6301 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     6361 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     6421 taaaaatgaa gttttaaatc aagcccaatc tgaataatgt tacaaccaat taaccaattc
     6481 tgattagaaa aactcatcga gcatcaaatg aaactgcaat ttattcatat caggattatc
     6541 aataccatat ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt
     6601 ccataggatg gcaagatcct ggtatcggtc tgcgattccg actcgtccaa catcaataca
     6661 acctattaat ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac
     6721 gactgaatcc ggtgagaatg gcaaaagttt atgcatttct ttccagactt gttcaacagg
     6781 ccagccatta cgctcgtcat caaaatcact cgcatcaacc aaaccgttat tcattcgtga
     6841 ttgcgcctga gcgagacgaa atacgcgatc gctgttaaaa ggacaattac aaacaggaat
     6901 cgaatgcaac cggcgcagga acactgccag cgcatcaaca atattttcac ctgaatcagg
     6961 atattcttct aatacctgga atgctgtttt tccggggatc gcagtggtga gtaaccatgc
     7021 atcatcagga gtacggataa aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca
     7081 gtttagtctg accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag
     7141 aaacaactct ggcgcatcgg gcttcccata caagcgatag attgtcgcac ctgattgccc
     7201 gacattatcg cgagcccatt tatacccata taaatcagca tccatgttgg aatttaatcg
     7261 cggcctcgac gtttcccgtt gaatatggct cataacaccc cttgtattac tgtttatgta
     7321 agcagacagt tttattgttc atgatgatat atttttatct tgtgcaatgt aacatcagag
     7381 attttgagac acgggccaga gctgca
//
