LOCUS       sequence-216996-        4088 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  An entry vector with U6a and U6c promoter driving guide RNAs 
            expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4088)
  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 4088)
  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
FEATURES             Location/Qualifiers
     source          1..4088
                     /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(2197..2292)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2329..2348)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2330..2348)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2353..2369)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2353..2369)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2471..2490)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2482..3291
                     /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(2549..2568)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3438..4026
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3927..3946
                     /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 atgctgtttc cagcatagct cttaaacaga
     1201 gacggtcgac agtctgcagt gtcgtctctc gaactaggag cctggagaac tgctatataa
     1261 acccgggact tctgggtatg tttctgccat gtggtaagtg gataaaccta cttattcaac
     1321 tcatttacct caaacatgac agtaaacctc ttcggggcaa acctgcaaac gcttaaaacc
     1381 ttcaaagttt gaaaccagca gttttgtaga ggtttaccgt agtatttttg aactgtaaag
     1441 catatgcaga ggaagctggt tttaggcttg tgtgattgga tgagttcttc tgtgacgtaa
     1501 ctgatctttt gtcctcgtgt ccttcgcctt cccccacagg catgcgcgga ccttcccccg
     1561 ccgcagacca gaggacaaca tgctcagacg ccagcaaacg gcccatcgaa acattcacag
     1621 actctcggat tggagagacc gggcccgagg tctctgacta aaaaagcacc gactcggtgc
     1681 cactttttca agttgataac ggactagcct tatttaaact tgctatgctg tttccagcat
     1741 agctcttaaa cagagacggt cgacagtctg cagtgtcgtc tctcgaacca agagctggag
     1801 ggagagctat atataccagg gacttctggg tatgtttttg ggaggtggtg agtgactaaa
     1861 ccacttattc agctccctta gatcaagtct gaccctatat catggtgaca taaacctgca
     1921 aactgataaa acctgaagga tctcaaatcc agagtttgtg tgagggatta ccgtggtaat
     1981 tttcaaactg taaagcatat gcaaaattat ctggtcttgg cttgagtgat tgggtgtctc
     2041 ggtgtgatgc agggacgttt tcagtgacgt gtccttctcc ctcccccaca ggcatgcgca
     2101 gaacatttcc cccctccttg aagaccagaa caaaagacgc cgagagcagg aaactcgtct
     2161 tactgaatga ccgaggctgg agaaatcgac cctaggcaac tttattatac aaagttggca
     2221 ttataaaaaa gcattgctta tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata
     2281 aaatcattat ttggagctcc atggtagcgt taacgcggcc gcgatatccc ctatagtgag
     2341 tcgtattaca tggtcatagc tgtttcctgg cagctctggc ccgtgtctca aaatctctga
     2401 tgttacattg cacaagataa aaatatatca tcatgaacaa taaaactgtc tgcttacata
     2461 aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcgag gccgcgatta
     2521 aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa
     2581 tcaggtgcga caatctatcg cttgtatggg aagcccgatg cgccagagtt gtttctgaaa
     2641 catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg
     2701 acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg
     2761 ttactcacca ctgcgatccc cggaaaaaca gcattccagg tattagaaga atatcctgat
     2821 tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca ttcgattcct
     2881 gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga
     2941 atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt
     3001 gaacaagtct ggaaagaaat gcataaactt ttgccattct caccggattc agtcgtcact
     3061 catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt
     3121 gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc
     3181 ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat
     3241 cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta atcagaattg
     3301 gttaattggt tgtaacactg gcagagcatt acgctgactt gacgggacgg cgcaagctca
     3361 tgaccaaaat cccttaacgt gagttacgcg tcgttccact gagcgtcaga ccccgtagaa
     3421 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     3481 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     3541 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg
     3601 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     3661 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     3721 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     3781 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagca ttgagaaagc
     3841 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     3901 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     3961 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     4021 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     4081 cacatgtt
//
