LOCUS       sequence-370294-        4601 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pTarget containing protospacer for AaCas12b 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_PS_VB
FEATURES             Location/Qualifiers
     source          1..4601
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(111..130)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      924..1512
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1413..1432
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1666..1683
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    1698..1837
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(1763..1782)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1923..1945
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1939..2130)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(4303..4321)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4389..4493
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4494..4562,4563..4601,1..753)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
ORIGIN
        1 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
       61 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
      121 gcggtattat cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat acactattct
      181 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
      241 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
      301 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
      361 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
      421 gacaccacga tgcctgcagc aatggcaaca acgttgcgca aactattaac tggcgaacta
      481 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
      541 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
      601 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
      661 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
      721 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
      781 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
      841 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
      901 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
      961 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     1021 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
     1081 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     1141 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     1201 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     1261 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     1321 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     1381 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     1441 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     1501 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     1561 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     1621 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     1681 gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     1741 caccgcaatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtata
     1801 cactccgcta tcgctacgtg actgggtcat ggctgcgccc cgacacccgc caacacccgc
     1861 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     1921 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgaggcagct
     1981 gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct gttcatccgc
     2041 gtccagctcg ttgagtttct ccagaagcgt taatgtctgg cttctgataa agcgggccat
     2101 gttaagggcg gttttttcct gtttggtcac tgatgcctcc gtgtaagggg gatttctgtt
     2161 catgggggta atgataccga tgaaacgaga gaggatgctc acgatacggg ttactgatga
     2221 tgaacatgcc cggttactgg aacgttgtga gggtaaacaa ctggcggtat ggatgcggcg
     2281 ggaccagaga aaaatcactc agggtcaatg ccagcgcttc gttaatacag atgtaggtgt
     2341 tccacagggt agccagcagc atcctgcgat gcagatccgg aacataatgg tgcagggcgc
     2401 tgacttccgc gtttccagac tttacgaaac acggaaaccg aagaccattc atgttgttgc
     2461 tcaggtcgca gacgttttgc agcagcagtc gcttcacgtt cgctcgcgta tcggtgattc
     2521 attctgctaa ccagtaaggc aaccccgcca gcctagccgg gtcctcaacg acaggagcac
     2581 gatcatgcgc acccgtggcc aggacccaac gctgcccgag atgcgccgcg tgcggctgct
     2641 ggagatggcg gacgcgatgg atatgttctg ccaagggttg gtttgcgcat tcacagttct
     2701 ccgcaagaat tgattggctc caattcttgg agtggtgaat ccgttagcga ggtgccgccg
     2761 gcttccattc aggtcgaggt ggcccggctc catgcaccgc gacgcaacgc ggggaggcag
     2821 acaaggtata gggcggcgcc tacaatccat gccaacccgt tccatgtgct cgccgaggcg
     2881 gcataaatcg ccgtgacgat cagcggtcca atgatcgaag ttaggctggt aagagccgcg
     2941 agcgatcctt gaagctgtcc ctgatggtcg tcatctacct gcctggacag catggcctgc
     3001 aacgcgggca tcccgatgcc gccggaagcg agaagaatca taatggggaa ggccatccag
     3061 cctcgcgtcg cgaacgccag caagacgtag cccagcgcgt cggccgccat gccggcgata
     3121 atggcctgct tctcgccgaa acgtttggtg gcgggaccag tgacgaaggc ttgagcgagg
     3181 gcgtgcaaga ttccgaatac cgcaagcgac aggccgatca tcgtcgcgct ccagcgaaag
     3241 cggtcctcgc cgaaaatgac ccagagcgct gccggcacct gtcctacgag ttgcatgatc
     3301 actgatagat acaagagcca taagaacctc agatccttcc gtatttaagt ttggtaaagg
     3361 taaaaagacg aatgatgcat ccgccagtat gttctctagt gtggttcgtt gtttttgcgt
     3421 gagccatgag aacgaaccat tgagatcata cttactttgc atgtcactca aaaattttgc
     3481 ctcaaaactg gtgagctgaa tttttgcagt taaagcatcg tgtagtgttt ttcttagtcc
     3541 gttatgtagg taggaatctg atgtaatggt tgttggtatt ttgtcaccat tcatttttat
     3601 ctggttgttc tcaagttcgg ttacgagatc catttgtcta tctagttcaa cttggaaaat
     3661 caacgtatca gtcgggcggc ctcgcttatc aaccaccaat ttcatattgc tgtaagtgtt
     3721 taaatcttta cttattggtt tcaaaaccca ttggttaagc cttttaaact catggtagtt
     3781 attttcaagc attaacatga acttaaattc atcaaggcta atctctatat ttgccttgtg
     3841 agttttcttt tgtgttagtt cttttaataa ccactcataa atcctcatag agtatttgtt
     3901 ttcaaaagac ttaacatgtt ccagattata ttttatgaat ttttttaact ggaaaagata
     3961 aggcaatatc tcttcactaa aaactaattc taatttttcg cttgagaact tggcatagtt
     4021 tgtccactgg aaaatctcaa agcctttaac caaaggattc ctgatttcca cagttctcgt
     4081 catcagctct ctggttgctt tagctaatac accataagca ttttccctac tgatgttcat
     4141 catctgagcg tattggttat aagtgaacga taccgtccgt tctttccttg tagggttttc
     4201 aatcgtgggg ttgagtagtg ccacacagca taaaattagc ttggtttcat gctccgttaa
     4261 gtcatagcga ctaatcgcta gttcatttgc tttgacgaaa gggcctcgtg atacgcctat
     4321 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     4381 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     4441 tcatgagaca ataaccctga taaatgcttc aataacattg aaaaaggaag agtatgagta
     4501 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     4561 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga t
//
