LOCUS       sequence-334636-        2516 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PCR template vector for amplifying gRNAcore(EF)-tRNA(Q) fragment; 
            used together with pAC-U63-tgRNA-nlsBFP or pAC-U63-tgRNA-Gal80.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2516)
  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 2516)
  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: pTR(EF)-tRNA(Q)
FEATURES             Location/Qualifiers
     source          1..2516
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(757..776)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(758..776)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(781..797)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(781..797)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(899..918)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             910..1719
                     /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(977..996)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1866..2454
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2355..2374
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggccgct cttcacgggt ttaagagcta tgctggaaac
      601 agcatagcaa gtttaaataa ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg
      661 gtgccagcgc ggttccatgg tgtaatggtt agcactcagg actctgaatc ctgcgatccg
      721 agttcaaatc tcggtggaac ctgaggaact gatatcccct atagtgagtc gtattacatg
      781 gtcatagctg tttcctggca gctctggccc gtgtctcaaa atctctgatg ttacattgca
      841 caagataaaa atatatcatc atgaacaata aaactgtctg cttacataaa cagtaataca
      901 aggggtgtta tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa ttccaacatg
      961 gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc aggtgcgaca
     1021 atctatcgct tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt
     1081 agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac ggaatttatg
     1141 cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt actcaccact
     1201 gcgatccccg gaaaaacagc attccaggta ttagaagaat atcctgattc aggtgaaaat
     1261 attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt
     1321 ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat gaataacggt
     1381 ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga acaagtctgg
     1441 aaagaaatgc ataaactttt gccattctca ccggattcag tcgtcactca tggtgatttc
     1501 tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga tgttggacga
     1561 gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct cggtgagttt
     1621 tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc tgatatgaat
     1681 aaattgcagt ttcatttgat gctcgatgag tttttctaat cagaattggt taattggttg
     1741 taacactggc agagcattac gctgacttga cgggacggcg caagctcatg accaaaatcc
     1801 cttaacgtga gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     1861 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     1921 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     1981 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     2041 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     2101 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     2161 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     2221 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     2281 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     2341 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     2401 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     2461 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgtt
//
