LOCUS       sequence-334652-        8695 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  gRNA cloning vector containing a U6:3 promoter and a gRNA2.1 
            scaffold..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8695)
  AUTHORS   Koreman GT, Xu Y, Hu Q, Zhang Z, Allen SE, Wolfner MF, Wang B, Han C
  TITLE     Upgraded CRISPR/Cas9 tools for tissue-specific mutagenesis in 
            Drosophila.
  JOURNAL   Proc Natl Acad Sci U S A. 2021 Apr 6;118(14). pii: 2014255118. doi: 
            10.1073/pnas.2014255118.
  PUBMED    33782117
REFERENCE   2  (bases 1 to 8695)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2021-04-6"; volume: "118(14). pii: 
            2014255118. doi"; pages: " 10.1073/pnas.2014255118"
COMMENT     Sequence Label: pAC-U63-gRNA2.1
FEATURES             Location/Qualifiers
     source          1..8695
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    join(1..31,32..233)
                     /label=P element 3' end
                     /note="P element 3' end"
     primer_bind     complement(943..961)
                     /label=pCasper-F
                     /note="Drosophila mini-white gene, reverse primer"
     CDS             join(1413..1484,1886..2159,2234..2888,2950..3265,3469..3600,
                     3671..4285)
                     /codon_start=1
                     /gene="white"
                     /product="Drosophila white gene eye color pigment"
                     /label=mini-white
                     /note="This is a modified version of the white gene lacking
                     part of the first intron."
                     /translation="MGQEDQELLIRGGSKHPSAEHLNNGDSGAASQSCINQGFGQAKNY
                     GTLRPPSPPEDSGSGSGQLAENLTYAWHNMDIFGAVNQPGSGWRQLVNRTRGLFCNERH
                     IPAPRKHLLKNVCGVAYPGELLAVMGSSGAGKTTLLNALAFRSPQGIQVSPSGMRLLNG
                     QPVDAKEMQARCAYVQQDDLFIGSLTAREHLIFQAMVRMPRHLTYRQRVARVDQVIQEL
                     SLSKCQHTIIGVPGRVKGLSGGERKRLAFASEALTDPPLLICDEPTSGLDSFTAHSVVQ
                     VLKKLSQKGKTVILTIHQPSSELFELFDKILLMAEGRVAFLGTPSEAVDFFSYVGAQCP
                     TNYNPADFYVQVLAVVPGREIESRDRIAKICDNFAISKVARDMEQLLATKNLEKPLEQP
                     ENGYTYKATWFMQFRAVLWRSWLSVLKEPLLVKVRLIQTTMVAILIGLIFLGQQLTQVG
                     VMNINGAIFLFLTNMTFQNVFATINVFTSELPVFMREARSRLYRCDTYFLGKTIAELPL
                     FLTVPLVFTAIAYPMIGLRAGVLHFFNCLALVTLVANVSTSFGYLISCASSSTSMALSV
                     GPPVIIPFLLFGGFFLNSGSVPVYLKWLSYLSWFRYANEGLLINQWADVEPGEISCTSS
                     NTTCPSSGKVILETLNFSAADLPLDYVGLAILIVSFRVLAYLALRLRARRKE"
     misc_feature    join(5064..5618,5619..5649)
                     /label=P element 5' end
     rep_origin      complement(5884..6472)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5964..5983)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6643..7434,7435..7503))
                     /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     7266..7285
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7504..7608)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7676..7694
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7732..7754)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7854..7873
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     protein_bind    8083..8152
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
ORIGIN
        1 catgatgaaa taacataagg tggtcccgtc ggcaagagac atccacttaa cgtatgcttg
       61 caataagtgc gagtgaaagg aatagtattc tgagtgtcgt attgagtctg agtgagacag
      121 cgatatgatt gttgattaac ccttagcatg tccgtggggt ttgaattaac tcataatatt
      181 aattagacga aattattttt aaagttttat ttttaataat ttgcgagtac gcaaagcttg
      241 gctgcagtcg acgctagcat atgtttgctc acctgtgatt gctcctactc aaatacaaaa
      301 acatcaaatt ttctgtcaat aaagcatatt tatttatatt tattttacag gaaagaattc
      361 cttttaaagt gtattttaac ctataatgaa aaacgattaa aaaaaataca taaaataatt
      421 cgaaaatttt tgaatagccc aggttgataa aaattcattt catacgtttt ataacttatg
      481 cccctaagta ttttttgacc atagtgtttc aattctacat taattttaca gagtagaatg
      541 aaacgccacc tactcagcca agaggcgaaa aggttagctc gccaagcaga gagggcgcca
      601 gtgctcacta ctttttataa ttctcaactt ctttttccag actcagttcg tatatataga
      661 cctattttca atttaacgtc gcaaggaaga gcgaggctct tcttgtgttt aagagctatg
      721 ctggaaacag catagcaagt ttaaataagg ctagtccgtt atcaactttg ctggaaacag
      781 caaagtggca ccgagtcggt gctttttttg cctacctgga gcctgagagt tgttcaataa
      841 aataaaaatg tttcgttttt ttgctttcgc cagtatttat tatttttcat caatatgtat
      901 tcaatttggt atgtatttac tagtggccta attctagtat gtatgtaagt taataaaacc
      961 catttttgcg gaaagtagat aaaaaaaaca tttttttttt ttactgcact ggatatcatt
     1021 gaacttatct gatcagtttt aaatttactt cgatccaagg gtatttgatg taccaggttc
     1081 tttcgattac ctctcactca aaatgacatt ccactcaaag tcagcgctgt ttgcctcctt
     1141 ctctgtccac agaaatatcg ccgtctcttt cgccgctgcg tccgctatct ctttcgccac
     1201 cgtttgtagc gttacgtagc gtcaatgtcc gccttcagtt gcattttgtc agcggtttcg
     1261 tgacgaagct ccaagcggtt tacgccatca attaaacaca aagtgctgtg ccaaaactcc
     1321 tctcgcttct tatttttgtt tgttttttga gtgattgggg tggtgattgg ttttgggtgg
     1381 gtaagcaggg gaaagtgtga aaaatcccgg caatgggcca agaggatcag gagctattaa
     1441 ttcgcggagg cagcaaacac ccatctgccg agcatctgaa caatgtgagt agtacatgtg
     1501 catacatctt aagttcactt gatctatagg aactgcgatt gcaacatcaa attgtctgcg
     1561 gcgtgagaac tgcgacccac aaaaatccca aaccgcaatt gcacaaacaa atagtgacac
     1621 gaaacagatt attctggtag ctgttctcgc tatataagac aatttttgag atcatatcat
     1681 gatcaagaca tctaaaggca ttcattttcg actatattct tttttacaaa aaatataaca
     1741 accagatatt ttaagctgat cctagatgca caaaaaataa ataaaagtat aaacctactt
     1801 cgtaggatac ttcggggtac tttttgttcg gggttagatg agcataacgc ttgtagttga
     1861 tatttgagat cccctatcat tgcagggtga cagcggagcg gcttcgcaga gctgcattaa
     1921 ccagggcttc gggcaggcca aaaactacgg cacgctccgg ccacccagtc cgccggagga
     1981 ctccggttca gggagcggcc aactagccga gaacctcacc tatgcctggc acaatatgga
     2041 catctttggg gcggtcaatc agccgggctc cggatggcgg cagctggtca accggacacg
     2101 cggactattc tgcaacgagc gacacatacc ggcgcccagg aaacatttgc tcaagaacgg
     2161 tgagtttcta ttcgcagtcg gctgatctgt gtgaaatctt aataaagggt ccaattacca
     2221 atttgaaact cagtttgcgg cgtggcctat ccgggcgaac ttttggccgt gatgggcagt
     2281 tccggtgccg gaaagacgac cctgctgaat gcccttgcct ttcgatcgcc gcagggcatc
     2341 caagtatcgc catccgggat gcgactgctc aatggccaac ctgtggacgc caaggagatg
     2401 caggccaggt gcgcctatgt ccagcaggat gacctcttta tcggctccct aacggccagg
     2461 gaacacctga ttttccaagc catggtgcgg atgccacgac atctgaccta tcggcagcga
     2521 gtggcccgcg tggatcaggt gatccaggag ctttcgctca gcaaatgtca gcacacgatc
     2581 atcggtgtgc ccggcagggt gaaaggtctg tccggcggag aaaggaagcg tctggcattc
     2641 gcctccgagg ctctaaccga tccgccgctt ctgatctgcg atgagcccac ctccggactg
     2701 gactccttta ccgcccacag cgtcgtccag gtgctgaaga agctgtcgca gaagggcaag
     2761 accgtcatcc tgaccattca tcagccgtct tccgagctgt ttgagctctt tgacaagatc
     2821 cttctgatgg ccgagggcag ggtagctttc ttgggcactc ccagcgaagc cgtcgacttc
     2881 ttttcctagt gagttcgatg tgtttattaa gggtatctag tattacataa catctcaact
     2941 cctatccagc gtgggtgccc agtgtcctac caactacaat ccggcggact tttacgtaca
     3001 ggtgttggcc gttgtgcccg gacgggagat cgagtcccgt gatcggatcg ccaagatatg
     3061 cgacaatttt gccattagca aagtagcccg ggatatggag cagttgttgg ccaccaaaaa
     3121 tctggagaag ccactggagc agccggagaa tgggtacacc tacaaggcca cctggttcat
     3181 gcagttccgg gcggtcctgt ggcgatcctg gctgtcggtg ctcaaggaac cactcctcgt
     3241 aaaagtgcga cttattcaga caacggtgag tggttccagt ggaaacaaat gatataacgc
     3301 ttacaattct tggaaacaaa ttcgctagat tttagataga attgcctgat tccacaccct
     3361 tcttagtttt tttcaatgag atgtatagtt tatagttttg cagaagataa ataaatttca
     3421 tttaactcgc gaatattaat gagatgcgag taacatttta atttgcagat ggttgccatc
     3481 ttgattggcc tcatcttttt gggccaacaa ctcacgcaag tgggtgtgat gaatatcaac
     3541 ggagccatct tcctcttcct gaccaacatg acctttcaaa acgtctttgc cacgataaat
     3601 gtaagtcatg tttagaatac atttgcattt caataattta ctaactttct aatgaatcga
     3661 ttcgatttag gtgttcacct cagagctgcc agtttttatg agggaggccc gaagtcgact
     3721 ttatcgctgt gacacatact ttctgggcaa aacgattgcc gaattgccgc tttttctcac
     3781 agtgccactg gtcttcacgg cgattgccta tccgatgatc ggactgcggg ccggagtgct
     3841 gcacttcttc aactgcctgg cgctggtcac tctggtggcc aatgtgtcaa cgtccttcgg
     3901 atatctaata tcctgcgcca gctcctcgac ctcgatggcg ctgtctgtgg gtccgccggt
     3961 tatcatacca ttcctgctct ttggcggctt cttcttgaac tcgggctcgg tgccagtata
     4021 cctcaaatgg ttgtcgtacc tctcatggtt ccgttacgcc aacgagggtc tgctgattaa
     4081 ccaatgggcg gacgtggagc cgggcgaaat tagctgcaca tcgtcgaaca ccacgtgccc
     4141 cagttcgggc aaggtcatcc tggagacgct taacttctcc gccgccgatc tgccgctgga
     4201 ctacgtgggt ctggccattc tcatcgtgag cttccgggtg ctcgcatatc tggctctaag
     4261 acttcgggcc cgacgcaagg agtagccgac atatatccga aataactgct tgtttttttt
     4321 ttttaccatt attaccatcg tgtttactgt ttattgcccc ctcaaaaagc taatgtaatt
     4381 atatttgtgc caataaaaac aagatatgac ctatagaata caagtatttc cccttcgaac
     4441 atccccacaa gtagactttg gatttgtctt ctaaccaaaa gacttacaca cctgcatacc
     4501 ttacatcaaa aactcgttta tcgctacata aaacaccggg atatattttt tatatacata
     4561 cttttcaaat cgcgcgccct cttcataatt cacctccacc acaccacgtt tcgtagttgc
     4621 tctttcgctg tctcccaccc gctctccgca acacattcac cttttgttcg acgaccttgg
     4681 agcgactgtc gttagttccg cgcgattcgg ttcgctcaaa tggttccgag tggttcattt
     4741 cgtctcaata gaaattagta ataaatattt gtatgtacaa tttatttgct ccaatatatt
     4801 tgtatatatt tccctcacag ctatatttat tctaatttaa tattatgact ttttaaggta
     4861 attttttgtg acctgttcgg agtgattagc gttacaattt gaactgaaag tgacatccag
     4921 tgtttgttcc ttgtgtagat gcatctcaaa aaaatggtgg gcataatagt gttgtttata
     4981 tatatcaaaa ataacaacta taataataag aatacattta atttagaaaa tgcttggatt
     5041 tcactggaac tagaattaat tcggctgctg ctctaaacga cgcatttcgt actccaaagt
     5101 acgaattttt tccctcaagc tcttattttc attaaacaat gaacaggacc taacgcacag
     5161 tcacgttatt gtttacataa atgatttttt ttactattca aacttactct gtttgtgtac
     5221 tcccactggt atagccttct tttatctttt ctggttcagg ctctatcact ttactaggta
     5281 cggcatctgc gttgagtcgc ctccttttaa atgtctgacc ttttgcaggt gcagccttcc
     5341 actgcgaatc attaaagtgg gtatcacaaa tttgggagtt ttcaccaagg ctgcacccaa
     5401 ggctctgctc ccacaatttt ctcttaatag cacacttcgg cacgtgaatt aattttactc
     5461 cagtcacagc tttgcagcaa aatttgcaat atttcatttt tttttattcc acgtaagggt
     5521 taatgttttc aaaaaaaaat tcgtccgcac acaacctttc ctctcaacaa gcaaacgtgc
     5581 actgaattta agtgtatact tcggtaagct tcggctatcg acgggaccac cttatgttat
     5641 ttcatcatgg gccagaccca cgtagtccag cggcagatcg gcggcggaga agttaagcgt
     5701 ctccaggatg accttgcccg aactggggca cgtggtgttc gacgatgtgc agctaatttc
     5761 gcccggctcc acgtccgccc attggttaat cagcagaccc tcgttggcgt aacggaacca
     5821 tgagaggtac gacaaccatt tgaggtatac tggcaccgag cccgagttca agaagaaggc
     5881 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     5941 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     6001 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     6061 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     6121 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     6181 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     6241 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     6301 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     6361 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     6421 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     6481 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     6541 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     6601 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     6661 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     6721 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     6781 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     6841 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     6901 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     6961 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     7021 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     7081 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     7141 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     7201 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     7261 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     7321 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     7381 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     7441 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     7501 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     7561 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     7621 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     7681 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     7741 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     7801 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc
     7861 agagcagatt gtactgagag tgcaccatat cgacatgccc gccgtgaccg tcgagaaccc
     7921 gctgacgctg ccccgcgtat ccgcacccgc cgacgccgtc gcacgtcccg tgctcaccgt
     7981 gaccaccgcg cccagcggtt tcgagggcga gggcttcccg gtgcgccgcg cgttcgccgg
     8041 gatcaactac cgccacctcg acccgttcat catgatggac cagatgggtg aggtggagta
     8101 cgcgcccggg gagcccaagg gcacgccctg gcacccgcac cgcggcttcg agaccgtgac
     8161 ctacatcgtc gatatgcggt gtgaaatacc gcaccgaatc gcgcggaact aacgacagtc
     8221 gctccaaggt cgtcgaacaa aaggtgaatg tgttgcggag agcgggtggg agacagcgaa
     8281 agagcaacta cgaaacgtgg tgtggtggag gtgaattatg aagagggcgc gcgatttgaa
     8341 aagtatgtat ataaaaaata tatcccggtg ttttatgtag cgataaacga gtttttgatg
     8401 taaggtatgc aggtgtgtaa gtcttttggt tagaagacaa atccaaagtc tacttgtggg
     8461 gatgttcgaa ggggaaatac ttgtattcta taggtcatat cttgttttta ttggcacaaa
     8521 tataattaca ttagcttttt gagggggcaa taaacagtaa acacgatggt aataatggta
     8581 aaaaaaaaaa acaagcagtt atttcggata tatgtcggct actccttgcg tcgggcccga
     8641 agtcttagag ccagatatgc gagcacccgg aagctcacga tgagaatggc cagac
//
