LOCUS       sequence-212861-        3485 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  An entry vector with U6a promoter driving guide RNA expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3485)
  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 3485)
  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
FEATURES             Location/Qualifiers
     source          1..3485
                     /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(1594..1689)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1726..1745)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1727..1745)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1750..1766)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1750..1766)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1868..1887)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1879..2688
                     /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(1946..1965)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2835..3423
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3324..3343
                     /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 gttcatagtg 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 tcgaggtctc tgactaaaaa agcaccgact cggtgccact
     1081 ttttcaagtt gataacggac tagccttatt taaacttgct atgctgtttc cagcatagct
     1141 cttaaacaga gacggtcgac agtctgcagt gtcgtctctc gaaccaagag ctggagggag
     1201 agctatatat accagggact tctgggtatg tttttgggag gtggtgagtg actaaaccac
     1261 ttattcagct cccttagatc aagtctgacc ctatatcatg gtgacataaa cctgcaaact
     1321 gataaaacct gaaggatctc aaatccagag tttgtgtgag ggattaccgt ggtaattttc
     1381 aaactgtaaa gcatatgcaa aattatctgg tcttggcttg agtgattggg tgtctcggtg
     1441 tgatgcaggg acgttttcag tgacgtgtcc ttctccctcc cccacaggca tgcgcagaac
     1501 atttcccccc tccttgaaga ccagaacaaa agacgccgag agcaggaaac tcgtcttact
     1561 gaatgaccga ggctggagaa agtcgaccct aggcaacttt attatacaaa gttggcatta
     1621 taaaaaagca ttgcttatca atttgttgca acgaacaggt cactatcagt caaaataaaa
     1681 tcattatttg gagctccatg gtagcgttaa cgcggccgcg atatccccta tagtgagtcg
     1741 tattacatgg tcatagctgt ttcctggcag ctctggcccg tgtctcaaaa tctctgatgt
     1801 tacattgcac aagataaaaa tatatcatca tgaacaataa aactgtctgc ttacataaac
     1861 agtaatacaa ggggtgttat gagccatatt caacgggaaa cgtcgaggcc gcgattaaat
     1921 tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt cgggcaatca
     1981 ggtgcgacaa tctatcgctt gtatgggaag cccgatgcgc cagagttgtt tctgaaacat
     2041 ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa ctggctgacg
     2101 gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga tgcatggtta
     2161 ctcaccactg cgatccccgg aaaaacagca ttccaggtat tagaagaata tcctgattca
     2221 ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt
     2281 tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca atcacgaatg
     2341 aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg gcctgttgaa
     2401 caagtctgga aagaaatgca taaacttttg ccattctcac cggattcagt cgtcactcat
     2461 ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg ttgtattgat
     2521 gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg gaactgcctc
     2581 ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat tgataatcct
     2641 gatatgaata aattgcagtt tcatttgatg ctcgatgagt ttttctaatc agaattggtt
     2701 aattggttgt aacactggca gagcattacg ctgacttgac gggacggcgc aagctcatga
     2761 ccaaaatccc ttaacgtgag ttacgcgtcg ttccactgag cgtcagaccc cgtagaaaag
     2821 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     2881 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     2941 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     3001 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     3061 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     3121 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     3181 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     3241 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     3301 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     3361 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     3421 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     3481 atgtt
//
