LOCUS       sequence-384661-        5265 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes Cas1, Cas2, leader, repeat and one spacer from F. Novicida 
            (type V-A).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5265)
  AUTHORS   Wu WY, Jackson SA, Almendros C, Haagsma AC, Yilmaz S, Gort G, van 
            der Oost J, Brouns SJJ, Staals RHJ
  TITLE     Adaptation by Type V-A and V-B CRISPR-Cas Systems Demonstrates 
            Conserved Protospacer Selection Mechanisms Between Diverse 
            CRISPR-Cas Types.
  JOURNAL   CRISPR J. 2022 Aug;5(4):536-547. doi: 10.1089/crispr.2021.0150. Epub
            2022 Jul 12.
  PUBMED    35833800
REFERENCE   2  (bases 1 to 5265)
  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: 
            "10.1089/crispr.2021.0150"; journalName: "CRISPR J"; date: 
            "2022-08"; volume: "5"; issue: "4"; pages: "536-547"
COMMENT     Sequence Label: pCas-Delta-4_1_2_VA
FEATURES             Location/Qualifiers
     source          1..5265
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(16..35)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(17..35)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(40..56)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(40..56)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             487..1278
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      1371..1959
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1860..1879
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2113..2130
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2289..2316)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2408..2494)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2543..2565
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2557..2574
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2558..2574
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2590..2689
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(5165..5264)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 catccagctg atatccccta tagtgagtcg tattacatgg tcatagctgt ttcctggcag
       61 ctctggcccg tgtctcaaaa tctctgatgt tacattgcac aagataaaaa tatatcatca
      121 tgcctcctct agaccagcca ggacagaaat gcctcgactt cgctgctacc caaggttgcc
      181 gggtgacgca caccgtggaa acggatgaag gcacgaaccc agtggacata agcctgttcg
      241 gttcgtaagc tgtaatgcaa gtagcgtatg cgctcacgca actggtccag aaccttgacc
      301 gaacgcagcg gtggtaacgg cgcagtggcg gttttcatgg cttgttatga ctgttttttt
      361 ggggtacagt ctatgcctcg ggcatccaag cagcaagcgc gttacgccgt gggtcgatgt
      421 ttgatgttat ggagcagcaa cgatgttacg cagcagggca gtcgccctaa aacaaagtta
      481 aacattatga gggaagcggt gatcgccgaa gtatcgactc aactatcaga ggtagttggc
      541 gtcatcgagc gccatctcga accgacgttg ctggccgtac atttgtacgg ctccgcagtg
      601 gatggcggcc tgaagccrca cagtgatatt gatttgctgg ttacggtgac cgtaaggctt
      661 gatgaaacaa cgcggcgagc tttgatcaac gaccttttgg aaacttcggc ttcccctgga
      721 gagagcgaga ttctccgcgc tgtagaagtc accattgttg tgcacgacga catcattccg
      781 tggcgttatc cagctaagcg cgaactgcaa tttggagaat ggcagcgcaa tgacattctt
      841 gcaggtatct tcgagccagc cacgatcgac attgatctgg ctatcttgct gacaaaagca
      901 agagaacata gcgttgcctt ggtaggtcca gcggcggagg aactctttga tccggttcct
      961 gaacaggatc tatttgaggc gctaaatgaa accttaacgc tatggaactc gccgcccgac
     1021 tgggctggcg atgagcgaaa tgtagtgctt acgttgtccc gcatttggta cagcgcagta
     1081 accggcaaaa tcgcgccgaa ggatgtcgct gccgactggg caatggagcg cctgccggcc
     1141 cagtatcagc ccgtcatact tgaagctaga caggcttatc ttggacaaga agaagatcgc
     1201 ttggcctcgc gcgcagatca gttggaagaa tttgtccact acgtgaaagg cgagatcacc
     1261 aaggtagtcg gcaaataacc ctcgagccac ccatgaccaa aatcccttaa cgtgagttac
     1321 gcgtcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     1381 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     1441 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     1501 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     1561 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     1621 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     1681 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     1741 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     1801 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     1861 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     1921 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     1981 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     2041 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     2101 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     2161 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     2221 aaagcgggca gtgagcgcaa cgcaattaat acgcgtaccg ctagccagga agagtttgta
     2281 gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt agtttgatgc ctggcagttt
     2341 atggcgggcg tcctgcccgc caccctccgg gccgttgctt cacaacgttc aaatccgctc
     2401 ccggcggatt tgtcctactc aggagagcgt tcaccgacaa acaacagata aaacgaaagg
     2461 cccagtcttc cgactgagcc tttcgtttta tttgatgcct ggcagttccc tactctcgcg
     2521 ttaacgctag catggatgtt ttcccagtca cgacgttgta aaacgacggc cagtcttaag
     2581 ctcgggcccc aaataatgat tttattttga ctgatagtga cctgttcgtt gcaacaaatt
     2641 gatgagcaat gcttttttat aatgccaact ttgtacaaaa aagttggcat ggcaacgacc
     2701 agtcatgtcg agcatgagct cctctccaaa ttgattgatg agttaaaggt caaggccaac
     2761 tcagaccccg aggctgatgt cctggccggg cgcctgctcc accgccttaa ggccgagtca
     2821 gttacacaca cagtagccga atatctggag gtcttctctg acaaattcta cgatgaggaa
     2881 ttcttccaga tgcaccggga tgagctggag acccgagtct ctgctttcgc gcagagcccg
     2941 gcctacgagc gcatcgtctc cagcggctac ctgtcggccc tgcgctacta tgacacctat
     3001 ctgtatgtgg ggcgcagcgg gaagcaggag agtgtgcagc acttttacat gcggttagcc
     3061 ggcttctgtg cctcaaccac ctgcctctac gcgggtctca gggcagccct gcagcgggcc
     3121 aggccggaga ttgagagtga catggaggtg tttgattact actttgagca cctaacctcc
     3181 cagacggtgt gctgctccac gccctttatg cgctttgccg gggtggaaaa ctccactctg
     3241 gccagctgca tcctcaccac ccccgacctc agctccgagt gggacgtgac ccaggccctc
     3301 tataggcacc tggggcgcta cctctttcag cgagccgggg tgggtgtagg ggtgacgggg
     3361 gctggccagg atgggaaaca catcagcctc ctgatgagga tgatcaacag ccacgtggag
     3421 taccacaact atggctgcaa gaggccggtc agtgtggcgg cctacatgga gccctggcac
     3481 agccagattt tcaagttttt ggaaacgaag ctgccggaga accacgagag gtgcccgggc
     3541 atctttacgg ggctctttgt ccccgagctc ttcttcaagc tttttaggga cacgccctgg
     3601 tcggactggt acctgtttga ccccaaggac gccggggacc tggagaggct ctacggggag
     3661 gagtttgagc gcgagtacta tcggctggtg acagcgggca agttttgtgg gcgggtctcc
     3721 atcaagtccc tgatgttctc tatcgtcaac tgcgccgtca aggccggcag ccccttcatc
     3781 cttttgaagg aggcctgcaa cgcccacttt tggcgcgacc tgcagggcga ggccatgaac
     3841 gccgccaacc tgtgcgccga ggtgctgcag ccctcgagga agtctgtggc cacctgcaat
     3901 ctggccaaca tctgcctccc gcgctgcctg gtgaatgcgc ctctggcggt gcgggcacag
     3961 cgggccgaca cgcaggggga tgaactcctg ctggccctcc ctcgactctc agtcacccta
     4021 cctggagagg gggcagtcgg tgatggattc tcgctagccc gcctcagaga tgccacccag
     4081 tgtgccacct ttgtggtggc ctgctccatt cttcagggat cccccactta tgattccagg
     4141 gatatggcct ccatgggcct cggggtgcag ggcctggccg atgtctttgc ggacctgggc
     4201 tggcagtaca ctgaccctcc ctctcgctcg ttaaacaagg aaatattcga acatatgtac
     4261 tttacggccc tctgcaccag tagtctgatt ggacttcaca ccaggaagat ttttccgggt
     4321 ttcaaacaga gcaagtatgc cggggggtgg tttcactggc acgattgggc aggaacagac
     4381 ctttctattc ccagggaaat ttggtctcgc ctctctgaac gcattgtgag ggatgggctt
     4441 ttcaattcac agtttatcgc cctgatgccc acctcaggct gtgcccaggt gacgggctgt
     4501 tcggacgcct tctacccctt ctatgccaat gcgtccacca aggtcaccaa caaggaggag
     4561 gcccttaggc caaaccggtc tttttggcgt catgtgcgtc tggatgacag ggaagctttg
     4621 aatcttgtcg ggggccgtgt ctcctgcctc ccggaggctc tgcggcagcg ctacctgcgt
     4681 ttccaaacgg cctttgatta caaccaggag gacctgattc agatgtcccg ggacagggcc
     4741 ccctttgtgg accagagcca atctcacagc ctgtttttgc gtgaggaaga tgccgcgcgg
     4801 gccagcacgc tagccaacct actggtgcgc agctacgagc tgggcctgaa gactatcatg
     4861 tactattgtc gcattgagaa ggccgccgat ctgggggtga tggagtgtaa ggccagcgcg
     4921 gctctgtcgg tgccgcggga ggaacagaat gagcggagtc ccgctgagca gatgccgcct
     4981 cgtcccatgg aaccggcgca ggttgcgggg ccggttgaca tcatgagcaa gggcccaggg
     5041 gagggaccag gtgggtggtg tgtgcccggg ggattggaag tgtgctataa gtaccgtcag
     5101 ctcttctcag aggatgatct gttggagact gacggtttta ctgaacgagc ctgtgaatct
     5161 tgccaaccaa ctttcttgta caaagttggc attataagaa agcattgctt atcaatttgt
     5221 tgcaacgaac aggtcactat cagtcaaaat aaaatcatta tttgc
//
