LOCUS       sequence-263491-        2145 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  itga9 sgRNA targeting to ""5'-TCCGCTGCCAGCCAGCCGG-3"" in pDR274.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2145)
  AUTHORS   Shin M, Nozaki T, Idrizi F, Isogai S, Ogasawara K, Ishida K, Yuge S,
            Roscoe B, Wolfe SA, Fukuhara S, Mochizuki N, Deguchi T, Lawson ND
  TITLE     Valves Are a Conserved Feature of the Zebrafish Lymphatic System.
  JOURNAL   Dev Cell. 2019 Nov 4;51(3):374-386.e5. doi: 
            10.1016/j.devcel.2019.08.019. Epub 2019 Sep 26.
  PUBMED    31564611
REFERENCE   2  (bases 1 to 2145)
  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"; doi: 
            "10.1016/j.devcel.2019.08.019"; journalName: "Dev Cell"; date: 
            "2019-11-4- 4"; volume: "51"; issue: "3"; pages: "374-386.e5"
COMMENT     Sequence Label: pDR-itga9_sgRNA4
FEATURES             Location/Qualifiers
     source          1..2145
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(22..39)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(22..38)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      79..666
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     567..586
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(678..705)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(729..1544)
                     /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="MSHIQRETSCLKPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNGDRVFRLAQAQSRMNNGLVGA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     1452..1471
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(1545..1647)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(1648..1679)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     primer_bind     1727..1743
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1727..1743
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        1872..1890
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_RNA        1910..1985
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
ORIGIN
        1 agaatcaagc tgactagata aactggccgt cgttttacac gggtgggcct ttcttcggta
       61 gaaaatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
      121 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
      181 tccgaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
      241 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
      301 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
      361 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
      421 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
      481 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
      541 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
      601 tttcgccacc tctgacttga gcatcgattt ttgtgatgct cgtcaggggg gcggagccta
      661 tggaaaaacg ccagcaacgc agaaaggccc acccgaaggt gagccaggtg attacattta
      721 ggtcctcatt agaaaaactc atcgagcatc aagtgaaact gcaatttatt catatcagga
      781 ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg
      841 cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg tccaacatca
      901 atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa atcaccatga
      961 gtgacgactg aatccggtga gaatggcaag agtttatgca tttctttcca gacttgttca
     1021 acaggccagc cattacgctc gtcatcaaaa tcactcgcac caaccaaacc gttattcatt
     1081 cgtgattgcg cctgagcgag acgaaatacg cgatcgccgt taaaaggaca attacaaaca
     1141 ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt ttcacctgaa
     1201 tcaggatatt cttctaatac ctggaatgct gttttccctg ggatcgcagt ggtgagtaac
     1261 catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat aaattccgtc
     1321 agccagttta gcctgaccat ctcatctgta acatcattgg caacgctacc tttgccatgt
     1381 ttcagaaaca actctggcgc atcgggcttc ccatacaatc gatagattgt cgcacctgat
     1441 tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat gttggaattt
     1501 aatcgcggct tcaagcaaga cgtttcccgt tgaatatggc tcattttagc ttccttagct
     1561 cctgaaaatc tcgataactc aaaaaatacg cccggtagtg atcttatttc attatggtga
     1621 aagttggaac ctcttacgtg ccgatcaagt caaaagcctc cggtcggagg cttttgactt
     1681 tctgctatgg aggtcaggta tgatttaaat ggtcagtatt gagcctcagg aaacagctat
     1741 gacatcaagc tgactagata atctagctga tcgtggaccg atcatacgta taatgccgta
     1801 agatcacggg tcgcagcaca gctcgcggtc cagtagtgat cgacactgct cgatccgctc
     1861 gcaccgctag ctaatacgac tcactatagg tccgctgcca gccagccggg ttttagagct
     1921 agaaatagca agttaaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc
     1981 ggtgctttta aaaagcttgg atcgacgaga gcagcgcgac tggatctgtc gcccgtctca
     2041 aacgcaaccc tccggcggtc gcatatcatt caggacgagc ctcagactcc agcgtaactg
     2101 gactgcaatc aactcactgg ctcaccttcc ggtccacgat cagct
//
