LOCUS       sequence-244214-        2757 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of gRNA targeting KDELC2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2757)
  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 2757)
  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"; journalName: 
            "Glycobiology. 2018 Jan 5. pii: 4791732. doi: 
            10.1093/glycob/cwx101."
COMMENT     Sequence Label: gh148
FEATURES             Location/Qualifiers
     source          1..2757
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..249
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1..21
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     172..191
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        270..345
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(503..520)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(613..1201)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(693..712)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1372..2163,2164..2232))
                     /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     1995..2014
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2233..2337)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2638..2660
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2652..2669
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2653..2669
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2680..2699
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2680..2698
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
       61 ataattggaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
      121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
      181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
      241 cgaaacaccg atttgactac gactttgaag ttttagagct agaaatagca agttaaaata
      301 aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt ttgtaccgaa
      361 ttcgaattaa cggtacctgg agcacaagac tggcctcatg ggccttccgc tcactgcccg
      421 ctttccagtc gggaaacctg tcgtgccagc tgcattaaca tggtcatagc tgtttccttg
      481 cgtattgggc gctctccgct tcctcgctca ctgactcgct gcgctcggtc gttcgggtaa
      541 agcctggggt gcctaatgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
      601 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
      661 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
      721 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
      781 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
      841 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
      901 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
      961 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     1021 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     1081 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     1141 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     1201 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     1261 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     1321 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     1381 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     1441 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     1501 aatgataccg cgagaaccac gctcaccggc tccagattta tcagcaataa accagccagc
     1561 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     1621 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     1681 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     1741 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     1801 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     1861 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     1921 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     1981 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     2041 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     2101 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     2161 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     2221 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     2281 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     2341 atttccccga aaagtgccac ctaaattgta agcgttaata ttttgttaaa attcgcgtta
     2401 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     2461 aaatcaaaag aatagaccga gatagggttg agtggccgct acagggcgct cccattcgcc
     2521 attcaggctg cgcaactgtt gggaagggcg tttcggtgcg ggcctcttcg ctattacgcc
     2581 agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc
     2641 agtcacgacg ttgtaaaacg acggccagtg agcgcgacgt aatacgactc actatagggc
     2701 gaattggcgg aaggccgtca aggccacgtg tcttgtccag agctcgatat cgaattc
//
