LOCUS       sequence-263492-        2145 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  itga9 sgRNA targeting to ""5'-CCCTCCAGCCTTCCATATG-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_sgRNA7
FEATURES             Location/Qualifiers
     source          1..2145
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     16..32
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     16..32
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        161..179
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_RNA        199..274
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(456..473)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(456..472)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      513..1100
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1001..1020
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1112..1139)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(1163..1978)
                     /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     1886..1905
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(1979..2081)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(2082..2113)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 gtcagtattg agcctcagga aacagctatg acatcaagct gactagataa tctagctgat
       61 cgtggaccga tcatacgtat aatgccgtaa gatcacgggt cgcagcacag ctcgcggtcc
      121 agtagtgatc gacactgctc gatccgctcg caccgctagc taatacgact cactataggc
      181 cctccagcct tccatatggt tttagagcta gaaatagcaa gttaaaataa ggctagtccg
      241 ttatcaactt gaaaaagtgg caccgagtcg gtgcttttaa aaagcttgga tcgacgagag
      301 cagcgcgact ggatctgtcg cccgtctcaa acgcaaccct ccggcggtcg catatcattc
      361 aggacgagcc tcagactcca gcgtaactgg actgcaatca actcactggc tcaccttccg
      421 gtccacgatc agctagaatc aagctgacta gataaactgg ccgtcgtttt acacgggtgg
      481 gcctttcttc ggtagaaaat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
      541 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
      601 ctaccaactc tttttccgag gtaactggct tcagcagagc gcagatacca aatactgttc
      661 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
      721 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
      781 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
      841 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
      901 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
      961 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     1021 atagtcctgt cgggtttcgc cacctctgac ttgagcatcg atttttgtga tgctcgtcag
     1081 gggggcggag cctatggaaa aacgccagca acgcagaaag gcccacccga aggtgagcca
     1141 ggtgattaca tttaggtcct cattagaaaa actcatcgag catcaagtga aactgcaatt
     1201 tattcatatc aggattatca ataccatatt tttgaaaaag ccgtttctgt aatgaaggag
     1261 aaaactcacc gaggcagttc cataggatgg caagatcctg gtatcggtct gcgattccga
     1321 ctcgtccaac atcaatacaa cctattaatt tcccctcgtc aaaaataagg ttatcaagtg
     1381 agaaatcacc atgagtgacg actgaatccg gtgagaatgg caagagttta tgcatttctt
     1441 tccagacttg ttcaacaggc cagccattac gctcgtcatc aaaatcactc gcaccaacca
     1501 aaccgttatt cattcgtgat tgcgcctgag cgagacgaaa tacgcgatcg ccgttaaaag
     1561 gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa
     1621 tattttcacc tgaatcagga tattcttcta atacctggaa tgctgttttc cctgggatcg
     1681 cagtggtgag taaccatgca tcatcaggag tacggataaa atgcttgatg gtcggaagag
     1741 gcataaattc cgtcagccag tttagcctga ccatctcatc tgtaacatca ttggcaacgc
     1801 tacctttgcc atgtttcaga aacaactctg gcgcatcggg cttcccatac aatcgataga
     1861 ttgtcgcacc tgattgcccg acattatcgc gagcccattt atacccatat aaatcagcat
     1921 ccatgttgga atttaatcgc ggcttcaagc aagacgtttc ccgttgaata tggctcattt
     1981 tagcttcctt agctcctgaa aatctcgata actcaaaaaa tacgcccggt agtgatctta
     2041 tttcattatg gtgaaagttg gaacctctta cgtgccgatc aagtcaaaag cctccggtcg
     2101 gaggcttttg actttctgct atggaggtca ggtatgattt aaatg
//
