LOCUS       sequence-334635-        2869 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PCR template vector for amplifying gRNAcore(2.1)-pU6.3 promoter 
            fragment; used together with pAC-CR7T-gRNA2.1-nlsBFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2869)
  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 2869)
  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: pGC(2.1)-U6.3
FEATURES             Location/Qualifiers
     source          1..2869
                     /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"
     promoter        673..1093
                     /label=dU6-3 promoter
                     /note="RNA polymerase III promoter for Drosophila U6-3 
                     snRNA (Port et al., 2014)"
     primer_bind     complement(1110..1129)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1111..1129)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1134..1150)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1134..1150)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1252..1271)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1263..2072
                     /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(1330..1349)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2219..2807
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2708..2727
                     /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 cttctgttta agagctatgc tggaaacagc
      601 atagcaagtt taaataaggc tagtccgtta tcaactttgc tggaaacagc aaagtggcac
      661 cgagtcggtg ctttttttgc tcacctgtga ttgctcctac tcaaatacaa aaacatcaaa
      721 ttttctgtca ataaagcata tttatttata tttattttac aggaaagaat tccttttaaa
      781 gtgtatttta acctataatg aaaaacgatt aaaaaaaata cataaaataa ttcgaaaatt
      841 tttgaatagc ccaggttgat aaaaattcat ttcatacgtt ttataactta tgcccctaag
      901 tattttttga ccatagtgtt tcaattctac attaatttta cagagtagaa tgaaacgcca
      961 cctactcagc caagaggcga aaaggttagc tcgccaagca gagagggcgc cagtgctcac
     1021 tactttttat aattctcaac ttctttttcc agactcagtt cgtatatata gacctatttt
     1081 caatttaacg tcgggaagag cctgatatcc cctatagtga gtcgtattac atggtcatag
     1141 ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg atgttacatt gcacaagata
     1201 aaaatatatc atcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg
     1261 ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt aaattccaac atggatgctg
     1321 atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc
     1381 gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg
     1441 ccaatgatgt tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc
     1501 cgaccatcaa gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc
     1561 ccggaaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg
     1621 atgcgctggc agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta
     1681 acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg
     1741 atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa
     1801 tgcataaact tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg
     1861 ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa
     1921 tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt
     1981 cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc
     2041 agtttcattt gatgctcgat gagtttttct aatcagaatt ggttaattgg ttgtaacact
     2101 ggcagagcat tacgctgact tgacgggacg gcgcaagctc atgaccaaaa tcccttaacg
     2161 tgagttacgc gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     2221 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     2281 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     2341 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     2401 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     2461 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     2521 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     2581 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     2641 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     2701 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     2761 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     2821 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgtt
//
