LOCUS       sequence-263474-        2145 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  itga9 sgRNA targeting to ""5'-GAATGACGGAGCTCTCTACG-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_sgRNA1
FEATURES             Location/Qualifiers
     source          1..2145
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     93..112
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        93..111
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_RNA        131..206
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(388..405)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(388..404)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      445..1032
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     933..952
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1044..1071)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(1095..1910)
                     /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     1818..1837
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(1911..2013)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(2014..2045)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     primer_bind     2093..2109
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2093..2109
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 gatcatacgt ataatgccgt aagatcacgg gtcgcagcac agctcgcggt ccagtagtga
       61 tcgacactgc tcgatccgct cgcaccgcta gctaatacga ctcactatag ggatgacgga
      121 gctctctacg gttttagagc tagaaatagc aagttaaaat aaggctagtc cgttatcaac
      181 ttgaaaaagt ggcaccgagt cggtgctttt aaaaagcttg gatcgacgag agcagcgcga
      241 ctggatctgt cgcccgtctc aaacgcaacc ctccggcggt cgcatatcat tcaggacgag
      301 cctcagactc cagcgtaact ggactgcaat caactcactg gctcaccttc cggtccacga
      361 tcagctagaa tcaagctgac tagataaact ggccgtcgtt ttacacgggt gggcctttct
      421 tcggtagaaa atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
      481 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
      541 tctttttccg aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
      601 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
      661 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
      721 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
      781 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
      841 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
      901 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
      961 gtcgggtttc gccacctctg acttgagcat cgatttttgt gatgctcgtc aggggggcgg
     1021 agcctatgga aaaacgccag caacgcagaa aggcccaccc gaaggtgagc caggtgatta
     1081 catttaggtc ctcattagaa aaactcatcg agcatcaagt gaaactgcaa tttattcata
     1141 tcaggattat caataccata tttttgaaaa agccgtttct gtaatgaagg agaaaactca
     1201 ccgaggcagt tccataggat ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca
     1261 acatcaatac aacctattaa tttcccctcg tcaaaaataa ggttatcaag tgagaaatca
     1321 ccatgagtga cgactgaatc cggtgagaat ggcaagagtt tatgcatttc tttccagact
     1381 tgttcaacag gccagccatt acgctcgtca tcaaaatcac tcgcaccaac caaaccgtta
     1441 ttcattcgtg attgcgcctg agcgagacga aatacgcgat cgccgttaaa aggacaatta
     1501 caaacaggaa tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac aatattttca
     1561 cctgaatcag gatattcttc taatacctgg aatgctgttt tccctgggat cgcagtggtg
     1621 agtaaccatg catcatcagg agtacggata aaatgcttga tggtcggaag aggcataaat
     1681 tccgtcagcc agtttagcct gaccatctca tctgtaacat cattggcaac gctacctttg
     1741 ccatgtttca gaaacaactc tggcgcatcg ggcttcccat acaatcgata gattgtcgca
     1801 cctgattgcc cgacattatc gcgagcccat ttatacccat ataaatcagc atccatgttg
     1861 gaatttaatc gcggcttcaa gcaagacgtt tcccgttgaa tatggctcat tttagcttcc
     1921 ttagctcctg aaaatctcga taactcaaaa aatacgcccg gtagtgatct tatttcatta
     1981 tggtgaaagt tggaacctct tacgtgccga tcaagtcaaa agcctccggt cggaggcttt
     2041 tgactttctg ctatggaggt caggtatgat ttaaatggtc agtattgagc ctcaggaaac
     2101 agctatgaca tcaagctgac tagataatct agctgatcgt ggacc
//
