LOCUS       sequence-358592-        3872 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Single guide RNA cassette under U3 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3872)
  AUTHORS   Bieluszewski T, Sura W, Dziegielewski W, Bieluszewska A, Lachance C,
            Kabza M, Szymanska-Lejman M, Abram M, Wlodzimierz P, De Winne N, De 
            Jaeger G, Sadowski J, Cote J, Ziolkowski PA
  TITLE     NuA4 and H2A.Z control environmental responses and autotrophic 
            growth in Arabidopsis.
  JOURNAL   Nat Commun. 2022 Jan 12;13(1):277. doi: 10.1038/s41467-021-27882-5.
  PUBMED    35022409
REFERENCE   2  (bases 1 to 3872)
  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"; journalName: "Nat 
            Commun."; date: "2022-01-12"; volume: "13(1):277. doi"; pages: " 
            10.1038/s41467-021-27882-5"
COMMENT     Sequence Label: pJET1.2-U3
FEATURES             Location/Qualifiers
     source          1..3872
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(25..42)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(196..784)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(276..295)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(955..1746,1747..1815))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1578..1597
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1816..1920)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1988..2006
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2312..2330
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     2378..2395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2379..2395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2402..2420
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_RNA        2986..3061
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(3168..3184)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3168..3184)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(3221..3239)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3260..3276)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3260..3276)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        complement(join(3711..3717,3718..3735,3736..3741))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    3756..3777
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
       61 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
      121 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
      181 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
      241 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
      301 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
      361 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
      421 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
      481 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
      541 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
      601 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
      661 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
      721 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
      781 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
      841 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
      901 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
      961 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     1021 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     1081 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     1141 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     1201 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     1261 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     1321 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     1381 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     1441 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     1501 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     1561 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     1621 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     1681 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     1741 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     1801 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     1861 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     1921 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     1981 ctataaaaat aggcgtatca cgaggccgcc cctgcagccg aattatatta tttttgccaa
     2041 ataattttta acaaaagctc tgaagtcttc ttcatttaaa ttcttagatg atacttcatc
     2101 tggaaaattg tcccaattag tagcatcacg ctgtgagtaa gttctaaacc atttttttat
     2161 tgttgtatta tctctaatct tactactcga tgagttttcg gtattatctc tatttttaac
     2221 ttggagcagg ttccattcat tgtttttttc atcatagtga ataaaatcaa ctgctttaac
     2281 acttgtgcct gaacaccata tccatccggc gtaatacgac tcactatagg gagagcggcc
     2341 gccagatctt ccggatggct cgagtttttc agcaagatgt aaaacgacgg ccagtgaatt
     2401 gtaatacgac tcactatagg gcgaattggg taccggatcc aagtttcgtt gaaacaatag
     2461 gtgtcagtga tgaaatcatg aaatgcattc acattggttt gttgtgtgtt caggataatg
     2521 cagagagtag accaacaatg gcttctgttg ttgtaatgct caatgcaaat tctttcactt
     2581 taccgagacc ctcgcaaccc gctttttaca ccggagatgg agaatcaatt ctaagataca
     2641 ccaatcagat taatcatatt gcgtctttga atgatgttac aattaccgag ttcgatgctc
     2701 gttaactttg tgtgtaaaag acaaaaatga taatttctgt ttgcttgaat gttatacact
     2761 tcaaaatgtc aaatgacgtt tgaatttaaa attctttgtt ctaataaaac catattcttt
     2821 aattattggg cttgaacata ttggctttgt taacaattta ggcccattag cttatgattg
     2881 ctttttgtta cgaattttac gtcccacatc ggtgagctag tgaagaaacg ttttctttat
     2941 atatgcatag aaagcatcaa gggtcatcga gctcccgtga tccaggtttt agagctagaa
     3001 atagcaagtt aaaataaggc tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg
     3061 ctttttttgt cccttcgaag ggcaattctg cagatatcca tcacactggc ggccgctaga
     3121 ggtcgacggt atcgataagc ttgatatcga attcctgcag cccgggggat ccactagttc
     3181 tagaacggcc gccaccgcgg tggagctcca gcttttgttc cctttagtga gggttaattt
     3241 cgagcttggc gtaatcatgg tcatagctgt ttcctgatct ttctagaaga tctcctacaa
     3301 tattctcagc tgccatggaa aatcgatgtt cttcttttat tctctcaaga ttttcaggct
     3361 gtatattaaa acttatatta agaactatgc taaccacctc atcaggaacc gttgtaggtg
     3421 gcgtgggttt tcttggcaat cgactctcat gaaaactacg agctaaatat tcaatatgtt
     3481 cctcttgacc aactttattc tgcatttttt ttgaacgagg tttagagcaa gcttcaggaa
     3541 actgagacag gaattttatt aaaaatttaa attttgaaga aagttcaggg ttaatagcat
     3601 ccattttttg ctttgcaagt tcctcagcat tcttaacaaa agacgtctct tttgacatgt
     3661 ttaaagttta aacctcctgt gtgaaattat tatccgctca taattccaca cattatacga
     3721 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     3781 gcgttgcgct cactgccaat tgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     3841 tgaatcggcc aacgcgcggg gagaggcggt tt
//
