LOCUS       sequence-215034-        2520 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PCR template vector for amplifying gRNA(F+E)-OstRNA fragment.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2520)
  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 2520)
  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: pMGC
FEATURES             Location/Qualifiers
     source          1..2520
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(208..227)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(209..227)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(232..248)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(232..248)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(350..369)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             361..1170
                     /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(428..447)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1317..1905
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1806..1825
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2059..2076
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2235..2262)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2354..2440)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2489..2511
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2503..2520
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2504..2520
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 cttaagctcg ggccgctctt cacgggttta agagctatgc tggaaacagc atagcaagtt
       61 taaataaggc tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg caacaaagca
      121 ccagtggtct agtggtagaa tagtaccctg ccacggtaca gacccgggtt cgattcccgg
      181 ctggtgcagg aagaagagcc tgatatcccc tatagtgagt cgtattacat ggtcatagct
      241 gtttcctggc agctctggcc cgtgtctcaa aatctctgat gttacattgc acaagataaa
      301 aatatatcat catgaacaat aaaactgtct gcttacataa acagtaatac aaggggtgtt
      361 atgagccata ttcaacggga aacgtcgagg ccgcgattaa attccaacat ggatgctgat
      421 ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac aatctatcgc
      481 ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg tagcgttgcc
      541 aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat gcctcttccg
      601 accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac tgcgatcccc
      661 ggaaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa tattgttgat
      721 gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg tccttttaac
      781 agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa tgaataacgg tttggttgat
      841 gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg
      901 cataaacttt tgccattctc accggattca gtcgtcactc atggtgattt ctcacttgat
      961 aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg agtcggaatc
     1021 gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt ttctccttca
     1081 ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcag
     1141 tttcatttga tgctcgatga gtttttctaa tcagaattgg ttaattggtt gtaacactgg
     1201 cagagcatta cgctgacttg acgggacggc gcaagctcat gaccaaaatc ccttaacgtg
     1261 agttacgcgt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     1321 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     1381 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     1441 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     1501 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     1561 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     1621 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     1681 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     1741 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     1801 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     1861 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     1921 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     1981 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     2041 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     2101 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     2161 gactggaaag cgggcagtga gcgcaacgca attaatacgc gtaccgctag ccaggaagag
     2221 tttgtagaaa cgcaaaaagg ccatccgtca ggatggcctt ctgcttagtt tgatgcctgg
     2281 cagtttatgg cgggcgtcct gcccgccacc ctccgggccg ttgcttcaca acgttcaaat
     2341 ccgctcccgg cggatttgtc ctactcagga gagcgttcac cgacaaacaa cagataaaac
     2401 gaaaggccca gtcttccgac tgagcctttc gttttatttg atgcctggca gttccctact
     2461 ctcgcgttaa cgctagcatg gatgttttcc cagtcacgac gttgtaaaac gacggccagt
//
