LOCUS       sequence-221333-        2739 bp    DNA     circular SYN 15-APR-2025
DEFINITION  expression of gRNA targeting EXTL2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2739)
  AUTHORS   Narimatsu Y, Joshi HJ, Zhang Y, Gomes C, Chen YH, Lorenzetti F, 
            Furukawa S, Schjoldager K, Hansen L, Clausen H, Bennett EP, Wandall 
            HH
  TITLE     A validated gRNA library for CRISPR/Cas9 targeting of the human 
            glycosyltransferase genome.
  JOURNAL   Glycobiology. 2018 Jan 5. pii: 4791732. doi: 10.1093/glycob/cwx101.
  PUBMED    29315387
REFERENCE   2  (bases 1 to 2739)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Glycobiology. 2018 Jan 5. pii: 4791732. doi: 
            10.1093/glycob/cwx101."
COMMENT     Sequence Label: gh138
FEATURES             Location/Qualifiers
     source          1..2739
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     278..300
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     292..309
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     293..309
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     320..339
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        320..338
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        398..646
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     398..418
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     569..588
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        668..743
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(882..899)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(992..1580)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1072..1091)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1751..2542,2543..2611))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2374..2393
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2612..2716)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtggccgct acagggcgct cccattcgcc attcaggctg cgcaactgtt
      181 gggaagggcg tttcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
      241 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
      301 acggccagtg agcgcgacgt aatacgactc actatagggc gaattggcgg aaggccgtca
      361 aggccacgtg tcttgtccag agctcgatat cgaattcgag ggcctatttc ccatgattcc
      421 ttcatatttg catatacgat acaaggctgt tagagagata attggaatta atttgactgt
      481 aaacacaaag atattagtac aaaatacgtg acgtagaaag taataatttc ttgggtagtt
      541 tgcagtttta aaattatgtt ttaaaatgga ctatcatatg cttaccgtaa cttgaaagta
      601 tttcgatttc ttggctttat atatcttgtg gaaaggacga aacaccgcag ctaccagtaa
      661 taatacggtt ttagagctag aaatagcaag ttaaaataag gctagtccgt tatcaacttg
      721 aaaaagtggc accgagtcgg tgcttttttt ggtacctgga gcacaagact ggcctcatgg
      781 gccttccgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaacat
      841 ggtcatagct gtttccttgc gtattgggcg ctctccgctt cctcgctcac tgactcgctg
      901 cgctcggtcg ttcgggtaaa gcctggggtg cctaatgagc aaaaggccag caaaaggcca
      961 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     1021 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     1081 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     1141 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     1201 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     1261 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     1321 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     1381 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     1441 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     1501 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     1561 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     1621 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     1681 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     1741 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     1801 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     1861 catctggccc cagtgctgca atgataccgc gagaaccacg ctcaccggct ccagatttat
     1921 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     1981 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     2041 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     2101 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     2161 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     2221 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     2281 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     2341 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     2401 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     2461 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     2521 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     2581 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     2641 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     2701 aaataggggt tccgcgcaca tttccccgaa aagtgccac
//
