LOCUS       sequence-215033-        2872 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PCR template vector for amplifying gRNAcore-pU6.3 promoter fragment.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2872)
  AUTHORS   Poe AR, Wang B, Sapar ML, Ji H, Li K, Onabajo T, Fazliyeva R, Gibbs 
            M, Qiu Y, Hu Y, Han C
  TITLE     Robust CRISPR/Cas9-Mediated Tissue Specific Mutagenesis Reveals Gene
            Redundancy and Perdurance in Drosophila.
  JOURNAL   Genetics. 2018 Nov 30. pii: genetics.118.301736. doi: 
            10.1534/genetics.118.301736.
  PUBMED    30504366
REFERENCE   2  (bases 1 to 2872)
  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: "Genetics. 
            2018 Nov 30. pii: genetics.118.301736. doi: 
            10.1534/genetics.118.301736."
COMMENT     Sequence Label: pTGC-U6.2
FEATURES             Location/Qualifiers
     source          1..2872
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        112..511
                     /label=dU6-2 promoter
                     /note="RNA polymerase III promoter for Drosophila U6-2 
                     snRNA (Das et al., 1987)"
     primer_bind     complement(529..548)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(530..548)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(553..569)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(553..569)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(671..690)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             682..1491
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(749..768)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1638..2226
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2127..2146
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2380..2397
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2556..2583)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2675..2761)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2810..2832
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2824..2841
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2825..2841
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_RNA        join(2864..2872,1..67)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
ORIGIN
        1 ctagaaatag caagttaaaa taaggctagt ccgttatcaa cttgaaaaag tggcaccgag
       61 tcggtgcttt ttttgtcaca tctacaatcg ttttatatgt tctcttcttt cgttcgactt
      121 gcagcctgaa atacggcacg agtaggaaaa gccgagtcaa atgccgaatg cagagtctca
      181 ttacagcaca atcaactcaa gaaaaactcg acactttttt accatttgca cttaaatcct
      241 tttttattcg ttatgtatac tttttttggt ccctaaccaa aacaaaacca aactctctta
      301 gtcgtgcctc tatatttaaa actatcaatt tattatagtc aataaatcga actgtgtttt
      361 caacaaacga acaataggac actttgattc taaaggaaat tttgaaaatc ttaagcagag
      421 ggttcttaag accatttgcc aattcttata attctcaact gctctttcct gatgttgatc
      481 atttatatag gtatgttttc ctcaatactt cgggaagagc ctgatatccc ctatagtgag
      541 tcgtattaca tggtcatagc tgtttcctgg cagctctggc ccgtgtctca aaatctctga
      601 tgttacattg cacaagataa aaatatatca tcatgaacaa taaaactgtc tgcttacata
      661 aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcgag gccgcgatta
      721 aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa
      781 tcaggtgcga caatctatcg cttgtatggg aagcccgatg cgccagagtt gtttctgaaa
      841 catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg
      901 acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg
      961 ttactcacca ctgcgatccc cggaaaaaca gcattccagg tattagaaga atatcctgat
     1021 tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca ttcgattcct
     1081 gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga
     1141 atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt
     1201 gaacaagtct ggaaagaaat gcataaactt ttgccattct caccggattc agtcgtcact
     1261 catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt
     1321 gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc
     1381 ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat
     1441 cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta atcagaattg
     1501 gttaattggt tgtaacactg gcagagcatt acgctgactt gacgggacgg cgcaagctca
     1561 tgaccaaaat cccttaacgt gagttacgcg tcgttccact gagcgtcaga ccccgtagaa
     1621 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     1681 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     1741 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     1801 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     1861 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     1921 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     1981 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     2041 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     2101 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     2161 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     2221 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     2281 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     2341 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     2401 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     2461 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatacg
     2521 cgtaccgcta gccaggaaga gtttgtagaa acgcaaaaag gccatccgtc aggatggcct
     2581 tctgcttagt ttgatgcctg gcagtttatg gcgggcgtcc tgcccgccac cctccgggcc
     2641 gttgcttcac aacgttcaaa tccgctcccg gcggatttgt cctactcagg agagcgttca
     2701 ccgacaaaca acagataaaa cgaaaggccc agtcttccga ctgagccttt cgttttattt
     2761 gatgcctggc agttccctac tctcgcgtta acgctagcat ggatgttttc ccagtcacga
     2821 cgttgtaaaa cgacggccag tcttaagctc gggccgctct tctgttttag ag
//
