LOCUS       sequence-370295-        4601 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pTarget containing protospacer with a mismatch on position 1, for 
            FnCas12a targeting.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4601)
  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 4601)
  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: pTarget_priming_VA
FEATURES             Location/Qualifiers
     source          1..4601
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(101..120)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      914..1502
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1403..1422
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1656..1673
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    1688..1827
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(1753..1772)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1913..1935
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1929..2120)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(4293..4311)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4379..4483
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4484..4552,4553..4601,1..743)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTIASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
ORIGIN
        1 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
       61 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
      121 cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
      181 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
      241 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
      301 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
      361 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
      421 tgcctgcagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactatag
      481 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
      541 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
      601 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
      661 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
      721 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
      781 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
      841 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
      901 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
      961 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     1021 aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt
     1081 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     1141 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     1201 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     1261 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     1321 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     1381 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     1441 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     1501 aaaacgccgg caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     1561 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     1621 ctgatacagc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     1681 aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcaatg
     1741 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtata cactccgcta
     1801 tcgctacgtg actgggtcat ggctgcgccc cgacacccgc caacacccgc tgacgcgccc
     1861 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     1921 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc
     1981 tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct gttcatccgc gtccagctcg
     2041 ttgagtttct ccagaagcgt taatgtctgg cttctgataa agcgggccat gttaagggcg
     2101 gttttttcct gtttggtcac tgatgcctcc gtgtaagggg gatttctgtt catgggggta
     2161 atgataccga tgaaacgaga gaggatgctc acgatacggg ttactgatga tgaacatgcc
     2221 cggttactgg aacgttgtga gggtaaacaa ctggcggtat ggatgcggcg ggaccagaga
     2281 aaaatcactc agggtcaatg ccagcgcttc gttaatacag atgtaggtgt tccacagggt
     2341 agccagcagc atcctgcgat gcagatccgg aacataatgg tgcagggcgc tgacttccgc
     2401 gtttccagac tttacgaaac acggaaaccg aagaccattc atgttgttgc tcaggtcgca
     2461 gacgttttgc agcagcagtc gcttcacgtt cgctcgcgta tcggtgattc attctgctaa
     2521 ccagtaaggc aaccccgcca gcctagccgg gtcctcaacg acaggagcac gatcatgcgc
     2581 acccgtggcc aggacccaac gctgcccgag atgcgccgcg tgcggctgct ggagatggcg
     2641 gacgcgatgg atatgttctg ccaagggttg gtttgcgcat tcacagttct ccgcaagaat
     2701 tgattggctc caattcttgg agtggtgaat ccgttagcga ggtgccgccg gcttccattc
     2761 aggtcgaggt ggcccggctc catgcaccgc gacgcaacgc ggggaggcag acaaggtata
     2821 gggcggcgcc tacaatccat gccaacccgt tccatgtgct cgccgaggcg gcataaatcg
     2881 ccgtgacgat cagcggtcca atgatcgaag ttaggctggt aagagccgcg agcgatcctt
     2941 gaagctgtcc ctgatggtcg tcatctacct gcctggacag catggcctgc aacgcgggca
     3001 tcccgatgcc gccggaagcg agaagaatca taatggggaa ggccatccag cctcgcgtcg
     3061 cgaacgccag caagacgtag cccagcgcgt cggccgccat gccggcgata atggcctgct
     3121 tctcgccgaa acgtttggtg gcgggaccag tgacgaaggc ttgagcgagg gcgtgcaaga
     3181 ttccgaatac cgcaagcgac aggccgatca tcgtcgcgct ccagcgaaag cggtcctcgc
     3241 cgaaaatgac ccagagcgct gccggcacct gtcctacgag ttgcatgatc actgatagat
     3301 acaagagcca taagaacctc agatccttcc gtatttatgt ttggtaaagg taaaaagacg
     3361 aatgatgcat ccgccagtat gttctctagt gtggttcgtt gtttttgcgt gagccatgag
     3421 aacgaaccat tgagatcata cttactttgc atgtcactca aaaattttgc ctcaaaactg
     3481 gtgagctgaa tttttgcagt taaagcatcg tgtagtgttt ttcttagtcc gttatgtagg
     3541 taggaatctg atgtaatggt tgttggtatt ttgtcaccat tcatttttat ctggttgttc
     3601 tcaagttcgg ttacgagatc catttgtcta tctagttcaa cttggaaaat caacgtatca
     3661 gtcgggcggc ctcgcttatc aaccaccaat ttcatattgc tgtaagtgtt taaatcttta
     3721 cttattggtt tcaaaaccca ttggttaagc cttttaaact catggtagtt attttcaagc
     3781 attaacatga acttaaattc atcaaggcta atctctatat ttgccttgtg agttttcttt
     3841 tgtgttagtt cttttaataa ccactcataa atcctcatag agtatttgtt ttcaaaagac
     3901 ttaacatgtt ccagattata ttttatgaat ttttttaact ggaaaagata aggcaatatc
     3961 tcttcactaa aaactaattc taatttttcg cttgagaact tggcatagtt tgtccactgg
     4021 aaaatctcaa agcctttaac caaaggattc ctgatttcca cagttctcgt catcagctct
     4081 ctggttgctt tagctaatac accataagca ttttccctac tgatgttcat catctgagcg
     4141 tattggttat aagtgaacga taccgtccgt tctttccttg tagggttttc aatcgtgggg
     4201 ttgagtagtg ccacacagca taaaattagc ttggtttcat gctccgttaa gtcatagcga
     4261 ctaatcgcta gttcatttgc tttgacgaaa gggcctcgtg atacgcctat ttttataggt
     4321 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     4381 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     4441 ataaccctga taaatgcttc aataacattg aaaaaggaag agtatgagta ttcaacattt
     4501 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     4561 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt g
//
