LOCUS       sequence-255623-        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"
     rep_origin      36..623
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     524..543
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(635..662)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(686..1501)
                     /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     1409..1428
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(1502..1604)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(1605..1636)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     primer_bind     1684..1700
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1684..1700
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        1829..1847
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_RNA        1867..1942
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(2124..2141)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2124..2140)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tgggcctttc ttcggtagaa aatcaaagga tcttcttgag atcctttttt tctgcgcgta
       61 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
      121 gagctaccaa ctctttttcc gaggtaactg gcttcagcag agcgcagata ccaaatactg
      181 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
      241 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
      301 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
      361 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
      421 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
      481 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
      541 tttatagtcc tgtcgggttt cgccacctct gacttgagca tcgatttttg tgatgctcgt
      601 caggggggcg gagcctatgg aaaaacgcca gcaacgcaga aaggcccacc cgaaggtgag
      661 ccaggtgatt acatttaggt cctcattaga aaaactcatc gagcatcaag tgaaactgca
      721 atttattcat atcaggatta tcaataccat atttttgaaa aagccgtttc tgtaatgaag
      781 gagaaaactc accgaggcag ttccatagga tggcaagatc ctggtatcgg tctgcgattc
      841 cgactcgtcc aacatcaata caacctatta atttcccctc gtcaaaaata aggttatcaa
      901 gtgagaaatc accatgagtg acgactgaat ccggtgagaa tggcaagagt ttatgcattt
      961 ctttccagac ttgttcaaca ggccagccat tacgctcgtc atcaaaatca ctcgcaccaa
     1021 ccaaaccgtt attcattcgt gattgcgcct gagcgagacg aaatacgcga tcgccgttaa
     1081 aaggacaatt acaaacagga atcgaatgca accggcgcag gaacactgcc agcgcatcaa
     1141 caatattttc acctgaatca ggatattctt ctaatacctg gaatgctgtt ttccctggga
     1201 tcgcagtggt gagtaaccat gcatcatcag gagtacggat aaaatgcttg atggtcggaa
     1261 gaggcataaa ttccgtcagc cagtttagcc tgaccatctc atctgtaaca tcattggcaa
     1321 cgctaccttt gccatgtttc agaaacaact ctggcgcatc gggcttccca tacaatcgat
     1381 agattgtcgc acctgattgc ccgacattat cgcgagccca tttataccca tataaatcag
     1441 catccatgtt ggaatttaat cgcggcttca agcaagacgt ttcccgttga atatggctca
     1501 ttttagcttc cttagctcct gaaaatctcg ataactcaaa aaatacgccc ggtagtgatc
     1561 ttatttcatt atggtgaaag ttggaacctc ttacgtgccg atcaagtcaa aagcctccgg
     1621 tcggaggctt ttgactttct gctatggagg tcaggtatga tttaaatggt cagtattgag
     1681 cctcaggaaa cagctatgac atcaagctga ctagataatc tagctgatcg tggaccgatc
     1741 atacgtataa tgccgtaaga tcacgggtcg cagcacagct cgcggtccag tagtgatcga
     1801 cactgctcga tccgctcgca ccgctagcta atacgactca ctataggtcc gctgccagcc
     1861 agccgggttt tagagctaga aatagcaagt taaaataagg ctagtccgtt atcaacttga
     1921 aaaagtggca ccgagtcggt gcttttaaaa agcttggatc gacgagagca gcgcgactgg
     1981 atctgtcgcc cgtctcaaac gcaaccctcc ggcggtcgca tatcattcag gacgagcctc
     2041 agactccagc gtaactggac tgcaatcaac tcactggctc accttccggt ccacgatcag
     2101 ctagaatcaa gctgactaga taaactggcc gtcgttttac acggg
//
