LOCUS       sequence-216998-        4063 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  An entry vector with U6a and U6c promoter driving ndufs2 guide RNAs 
            expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4063)
  AUTHORS   Zhou W, Cao L, Jeffries J, Zhu X, Staiger CJ, Deng Q
  TITLE     Neutrophil-specific knockout demonstrates a role for mitochondria in
            regulating neutrophil motility in zebrafish.
  JOURNAL   Dis Model Mech. 2018 Mar 28;11(3). pii: dmm.033027. doi: 
            10.1242/dmm.033027.
  PUBMED    29590639
REFERENCE   2  (bases 1 to 4063)
  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: "Dis Model 
            Mech."; date: "2018-03-28"; volume: "11(3). pii: dmm.033027. doi"; 
            pages: " 10.1242/dmm.033027"
COMMENT     Sequence Label: p3E-U6a-U6c-ndufs2 guide
FEATURES             Location/Qualifiers
     source          1..4063
                     /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     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(624..747)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(761..777)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    793..927
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(817..836)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     871..890
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     protein_bind    complement(2172..2267)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2304..2323)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2305..2323)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2328..2344)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2328..2344)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2446..2465)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2457..3266
                     /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(2524..2543)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3413..4001
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3902..3921
                     /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 tcgggccctg cagctctaga gctcgaattc tacaggtcac
      601 taataccatc taagtagttg attcatagtg actgcatatg ttgtgtttta cagtattatg
      661 tagtctgttt tttatgcaaa atctaattta atatattgat atttatatca ttttacgttt
      721 ctcgttcaac tttcttgtac aaagttgaga tctataattc actggccgtc gttttacggt
      781 accatcgatg atgatccaga catgataaga tacattgatg agtttggaca aaccacaact
      841 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
      901 accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag
      961 gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtatg
     1021 gctgattatg atcctctaga tcccttgggt agcagcgaag tcgaggcatt tctgtcctgg
     1081 ctggtctaga ggtctctcag gtttaaaaaa agcaccgact cggtgccact ttttcaagtt
     1141 gataacggac tagccttatt taaacttgct atgctgtttt ccagcatagc tcttaaaccc
     1201 tcagcgccaa tgacacccga actaggagcc tggagaactg ctatataaac ccgggacttc
     1261 tgggtatgtt tctgccatgt ggtaagtgga taaacctact tattcaactc atttacctca
     1321 aacatgacag taaacctctt cggggcaaac ctgcaaacgc ttaaaacctt caaagtttga
     1381 aaccagcagt tttgtagagg tttaccgtag tatttttgaa ctgtaaagca tatgcagagg
     1441 aagctggttt taggcttgtg tgattggatg agttcttctg tgacgtaact gatcttttgt
     1501 cctcgtgtcc ttcgccttcc cccacaggca tgcgcggacc ttcccccgcc gcagaccaga
     1561 ggacaacatg ctcagacgcc agcaaacggc ccatcgaaac attcacagac tctcggattg
     1621 gagagaccgg gcccgaggtc tctgactaaa aaagcaccga ctcggtgcca ctttttcaag
     1681 ttgataacgg actagcctta tttaaacttg ctatgctgtt tccagcatag ctcttaaacc
     1741 cgggtctgac atatgcaccg aaccaagagc tggagggaga gctatatata ccagggactt
     1801 ctgggtatgt ttttgggagg tggtgagtga ctaaaccact tattcagctc ccttagatca
     1861 agtctgaccc tatatcatgg tgacataaac ctgcaaactg ataaaacctg aaggatctca
     1921 aatccagagt ttgtgtgagg gattaccgtg gtaattttca aactgtaaag catatgcaaa
     1981 attatctggt cttggcttga gtgattgggt gtctcggtgt gatgcaggga cgttttcagt
     2041 gacgtgtcct tctccctccc ccacaggcat gcgcagaaca tttcccccct ccttgaagac
     2101 cagaacaaaa gacgccgaga gcaggaaact cgtcttactg aatgaccgag gctggagaaa
     2161 tcgaccctag gcaactttat tatacaaagt tggcattata aaaaagcatt gcttatcaat
     2221 ttgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttgga gctccatggt
     2281 agcgttaacg cggccgcgat atcccctata gtgagtcgta ttacatggtc atagctgttt
     2341 cctggcagct ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata
     2401 tatcatcatg aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga
     2461 gccatattca acgggaaacg tcgaggccgc gattaaattc caacatggat gctgatttat
     2521 atgggtataa atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt
     2581 atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg
     2641 atgttacaga tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca
     2701 tcaagcattt tatccgtact cctgatgatg catggttact caccactgcg atccccggaa
     2761 aaacagcatt ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc
     2821 tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg
     2881 atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga
     2941 gtgattttga tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata
     3001 aacttttgcc attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc
     3061 ttatttttga cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag
     3121 accgatacca ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac
     3181 agaaacggct ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc
     3241 atttgatgct cgatgagttt ttctaatcag aattggttaa ttggttgtaa cactggcaga
     3301 gcattacgct gacttgacgg gacggcgcaa gctcatgacc aaaatccctt aacgtgagtt
     3361 acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     3421 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     3481 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     3541 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     3601 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     3661 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     3721 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     3781 aactgagata cctacagcgt gagcattgag aaagcgccac gcttcccgaa gggagaaagg
     3841 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     3901 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     3961 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     4021 ttttacggtt cctggccttt tgctggcctt ttgctcacat gtt
//
