LOCUS       sequence-216995-        3521 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  An entry vector with U6a promoter driving polg guide RNA expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3521)
  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 3521)
  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-polg guide
FEATURES             Location/Qualifiers
     source          1..3521
                     /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(1630..1725)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1762..1781)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1763..1781)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1786..1802)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1786..1802)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1904..1923)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1915..2724
                     /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(1982..2001)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2871..3459
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3360..3379
                     /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 cttaaactgc
     1201 gtagccacac caccatcgaa ccaagagctg gagggagagc tatatatacc agggacttct
     1261 gggtatgttt ttgggaggtg gtgagtgact aaaccactta ttcagctccc ttagatcaag
     1321 tctgacccta tatcatggtg acataaacct gcaaactgat aaaacctgaa ggatctcaaa
     1381 tccagagttt gtgtgaggga ttaccgtggt aattttcaaa ctgtaaagca tatgcaaaat
     1441 tatctggtct tggcttgagt gattgggtgt ctcggtgtga tgcagggacg ttttcagtga
     1501 cgtgtccttc tccctccccc acaggcatgc gcagaacatt tcccccctcc ttgaagacca
     1561 gaacaaaaga cgccgagagc aggaaactcg tcttactgaa tgaccgaggc tggagaaatc
     1621 gaccctaggc aactttatta tacaaagttg gcattataaa aaagcattgc ttatcaattt
     1681 gttgcaacga acaggtcact atcagtcaaa ataaaatcat tatttggagc tccatggtag
     1741 cgttaacgcg gccgcgatat cccctatagt gagtcgtatt acatggtcat agctgtttcc
     1801 tggcagctct ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga taaaaatata
     1861 tcatcatgaa caataaaact gtctgcttac ataaacagta atacaagggg tgttatgagc
     1921 catattcaac gggaaacgtc gaggccgcga ttaaattcca acatggatgc tgatttatat
     1981 gggtataaat gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgcttgtat
     2041 gggaagcccg atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat
     2101 gttacagatg agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc
     2161 aagcatttta tccgtactcc tgatgatgca tggttactca ccactgcgat ccccggaaaa
     2221 acagcattcc aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg
     2281 gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagcgat
     2341 cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt
     2401 gattttgatg acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataaa
     2461 cttttgccat tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt
     2521 atttttgacg aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac
     2581 cgataccagg atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag
     2641 aaacggcttt ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat
     2701 ttgatgctcg atgagttttt ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc
     2761 attacgctga cttgacggga cggcgcaagc tcatgaccaa aatcccttaa cgtgagttac
     2821 gcgtcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     2881 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     2941 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     3001 cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct
     3061 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     3121 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     3181 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     3241 ctgagatacc tacagcgtga gcattgagaa agcgccacgc ttcccgaagg gagaaaggcg
     3301 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     3361 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     3421 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     3481 ttacggttcc tggccttttg ctggcctttt gctcacatgt t
//
