LOCUS       sequence-370297-        4566 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  used as source for acquisition of new spacers.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4566)
  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 4566)
  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
FEATURES             Location/Qualifiers
     source          1..4566
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    16..155
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     71..90
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(170..187)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(341..929)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(421..440)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1100..1891,1892..1960))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLVARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1723..1742
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1961..2065)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2133..2151
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             4289..4480
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(4474..4496)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc gatagcggag tgtatactgg
       61 cttaactatg cggcatcaga gcagattgta ctgagagtgc accattgcgg tgtgaaatac
      121 cgcacagatg cgtaaggaga aaataccgca tcaggcgctc ttccgcttcc tcgctcactg
      181 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
      241 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
      301 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
      361 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
      421 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
      481 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
      541 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
      601 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
      661 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
      721 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
      781 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
      841 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
      901 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
      961 acgctaagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     1021 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     1081 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     1141 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     1201 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     1261 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     1321 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     1381 cagttaatag tttgcgcaac gttgttgcca ttgctgcagg catcgtagtg tcacgctcgt
     1441 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     1501 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     1561 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     1621 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     1681 gtatgcggcg accgagttgc tcttgcccgg cgtcaacacg ggataatacc gcgccacata
     1741 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     1801 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcaaccaac tgatcttcag
     1861 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     1921 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatgtt
     1981 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     2041 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag
     2101 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
     2161 aaagcaaatg aactagcgat tagtcgctat gacttaacgg agcatgaaac caagctaatt
     2221 ttatgctgtg tggcactact caaccccacg attgaaaacc ctacaaggaa agaacggacg
     2281 gtatcgttca cttataacca atacgctcag atgatgaaca tcagtaggga aaatgcttat
     2341 ggtgtattag ctaaagcaac cagagagctg atgacgagaa ctgtggaaat caggaatcct
     2401 ttggttaaag gctttgagat tttccagtgg acaaactatg ccaagttctc aagcgaaaaa
     2461 ttagaattag tttttagtga agagatattg ccttatcttt tccagttaaa aaaattcata
     2521 aaatataatc tggaacatgt taagtctttt gaaaacaaat actctatgag gatttatgag
     2581 tggttattaa aagaactaac acaaaagaaa actcacaagg caaatataga gattagcctt
     2641 gatgaattta agttcatgtt aatgcttgaa aataactacc atgagtttaa aaggcttaac
     2701 caatgggttt tgaaaccaat aagtaaagat ttaaacactt acagcaatat gaaattggtg
     2761 gttgataagc gaggccgccc gactgatacg ttgattttcc aagttgaact agatagacaa
     2821 atggatctcg taaccgaact tgagaacaac cagataaaaa tgaatggtga caaaatacca
     2881 acaaccatta catcagattc ctacctacat aacggactaa gaaaaacact acacgatgct
     2941 ttaactgcaa aaattcagct caccagtttt gaggcaaaat ttttgagtga catgcaaagt
     3001 aagtatgatc tcaatggttc gttctcatgg ctcacgcaaa aacaacgaac cacactagag
     3061 aacatactgg ctaaatacgg aaggatctga ggttcttatg gctcttgtat ctatcagtga
     3121 tcatgcaact cgtaggacag gtgccggcag cgctctgggt cattttcggc gaggaccgct
     3181 ttcgctggag cgcgacgatg atcggcctgt cgcttgcggt attcggaatc ttgcacgccc
     3241 tcgctcaagc cttcgtcact ggtcccgcca ccaaacgttt cggcgagaag caggccatta
     3301 tcgccggcat ggcggccgac gcgctgggct acgtcttgct ggcgttcgcg acgcgaggct
     3361 ggatggcctt ccccattatg attcttctcg cttccggcgg catcgggatg cccgcgttgc
     3421 aggccatgct gtccaggcag gtagatgacg accatcaggg acagcttcaa ggatcgctcg
     3481 cggctcttac cagcctaact tcgatcattg gaccgctgat cgtcacggcg atttatgccg
     3541 cctcggcgag cacatggaac gggttggcat ggattgtagg cgccgcccta taccttgtct
     3601 gcctccccgc gttgcgtcgc ggtgcatgga gccgggccac ctcgacctga atggaagccg
     3661 gcggcacctc gctaacggat tcaccactcc aagaattgga gccaatcaat tcttgcggag
     3721 aactgtgaat gcgcaaacca acccttggca gaacatatcc atcgcgtccg ccatctccag
     3781 cagccgcacg cggcgcatct cgggcagcgt tgggtcctgg ccacgggtgc gcatgatcgt
     3841 gctcctgtcg ttgaggaccc ggctaggctg gcggggttgc cttactggtt agcagaatga
     3901 atcaccgata cgcgagcgaa cgtgaagcga ctgctgctgc aaaacgtctg cgacctgagc
     3961 aacaacatga atggtcttcg gtttccgtgt ttcgtaaagt ctggaaacgc ggaagtcagc
     4021 gccctgcacc attatgttcc ggatctgcat cgcaggatgc tgctggctac cctgtggaac
     4081 acctacatct gtattaacga agcgctggca ttgaccctga gtgatttttc tctggtcccg
     4141 ccgcatccat accgccagtt gtttaccctc acaacgttcc agtaaccggg catgttcatc
     4201 atcagtaacc cgtatcgtga gcatcctctc tcgtttcatc ggtatcatta cccccatgaa
     4261 cagaaatccc ccttacacgg aggcatcagt gaccaaacag gaaaaaaccg cccttaacat
     4321 ggcccgcttt atcagaagcc agacattaac gcttctggag aaactcaacg agctggacgc
     4381 ggatgaacag gcagacatct gtgaatcgct tcacgaccac gctgatgagc tttaccgcag
     4441 ctgcctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac
     4501 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtaagg gcgcgtaagc
     4561 gggtgt
//
