LOCUS       sequence-370296-        4597 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pTarget containing protospacer for FnCas12a targeting.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4597)
  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 4597)
  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_VA
FEATURES             Location/Qualifiers
     source          1..4597
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(102..121)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      915..1503
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1404..1423
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1657..1674
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    1689..1828
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(1754..1773)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1914..1936
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1930..2121)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(4290..4308)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4376..4480
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4481..4549,4550..4597,1..744)
                     /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 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
       61 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
      121 tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
      181 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
      241 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
      301 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
      361 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
      421 atgcctgcag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
      481 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
      541 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
      601 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
      661 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
      721 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
      781 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
      841 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
      901 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
      961 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     1021 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     1081 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     1141 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     1201 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     1261 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     1321 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     1381 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     1441 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     1501 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     1561 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     1621 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     1681 gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcaat
     1741 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat acactccgct
     1801 atcgctacgt gactgggtca tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc
     1861 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     1921 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag
     1981 ctcatcagcg tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg cgtccagctc
     2041 gttgagtttc tccagaagcg ttaatgtctg gcttctgata aagcgggcca tgttaagggc
     2101 ggttttttcc tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt
     2161 aatgataccg atgaaacgag agaggatgct cacgatacgg gttactgatg atgaacatgc
     2221 ccggttactg gaacgttgtg agggtaaaca actggcggta tggatgcggc gggaccagag
     2281 aaaaatcact cagggtcaat gccagcgctt cgttaataca gatgtaggtg ttccacaggg
     2341 tagccagcag catcctgcga tgcagatccg gaacataatg gtgcagggcg ctgacttccg
     2401 cgtttccaga ctttacgaaa cacggaaacc gaagaccatt catgttgttg ctcaggtcgc
     2461 agacgttttg cagcagcagt cgcttcacgt tcgctcgcgt atcggtgatt cattctgcta
     2521 accagtaagg caaccccgcc agcctagccg ggtcctcaac gacaggagca cgatcatgcg
     2581 cacccgtggc caggacccaa cgctgcccga gatgcgccgc gtgcggctgc tggagatggc
     2641 ggacgcgatg gatatgttct gccaagggtt ggtttgcgca ttcacagttc tccgcaagaa
     2701 ttgattggct ccaattcttg gagtggtgaa tccgttagcg aggtgccgcc ggcttccatt
     2761 caggtcgagg tggcccggct ccatgcaccg cgacgcaacg cggggaggca gacaaggtat
     2821 agggcggcgc ctacaatcca tgccaacccg ttccatgtgc tcgccgaggc ggcataaatc
     2881 gccgtgacga tcagcggtcc aatgatcgaa gttaggctgg taagagccgc gagcgatcct
     2941 tgaagctgtc cctgatggtc gtcatctacc tgcctggaca gcatggcctg caacgcgggc
     3001 atcccgatgc cgccggaagc gagaagaatc ataatgggga aggccatcca gcctcgcgtc
     3061 gcgaacgcca gcaagacgta gcccagcgcg tcggccgcca tgccggcgat aatggcctgc
     3121 ttctcgccga aacgtttggt ggcgggacca gtgacgaagg cttgagcgag ggcgtgcaag
     3181 attccgaata ccgcaagcga caggccgatc atcgtcgcgc tccagcgaaa gcggtcctcg
     3241 ccgaaaatga cccagagcgc tgcaggcacc tgtcctacga gttgcatgat cactgataga
     3301 tacaagagcc ataagaacct cagatccttc cgtatttaga gaagtcattt aataaggcca
     3361 ctgttaaaag ccagtatgtt ctctagtgtg gttcgttgtt tttgcgtgag ccatgagaac
     3421 gaaccattga gatcatactt actttgcatg tcactcaaaa attttgcctc aaaactggtg
     3481 agctgaattt ttgcagttaa agcatcgtgt agtgtttttc ttagtccgtt atgtaggtag
     3541 gaatctgatg taatggttgt tggtattttg tcaccattca tttttatctg gttgttctca
     3601 agttcggtta cgagatccat ttgtctatct agttcaactt ggaaaatcaa cgtatcagtc
     3661 gggcggcctc gcttatcaac caccaatttc atattgctgt aagtgtttaa atctttactt
     3721 attggtttca aaacccattg gttaagcctt ttaaactcat ggtagttatt ttcaagcatt
     3781 aacatgaact taaattcatc aaggctaatc tctatatttg ccttgtgagt tttcttttgt
     3841 gttagttctt ttaataacca ctcataaatc ctcatagagt atttgttttc aaaagactta
     3901 acatgttcca gattatattt tatgaatttt tttaactgga aaagataagg caatatctct
     3961 tcactaaaaa ctaattctaa tttttcgctt gagaacttgg catagtttgt ccactggaaa
     4021 atctcaaagc ctttaaccaa aggattcctg atttccacag ttctcgtcat cagctctctg
     4081 gttgctttag ctaatacacc ataagcattt tccctactga tgttcatcat ctgagcgtat
     4141 tggttataag tgaacgatac cgtccgttct ttccttgtag ggttttcaat cgtggggttg
     4201 agtagtgcca cacagcataa aattagcttg gtttcatgct ccgttaagtc atagcgacta
     4261 atcgctagtt catttgcttt gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     4321 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atatgcgcgg
     4381 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     4441 accctgataa atgcttcaat aacattgaaa aaggaagagt atgagtattc aacatttccg
     4501 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     4561 gctggtgaaa gtaaaagatg ctgaagatca gttgggt
//
