LOCUS       sequence-154466-        2262 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  hU6 expression of gRNA NT1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2262)
  AUTHORS   Chaikind B, Bessen JL, Thompson DB, Hu JH, Liu DR
  TITLE     A programmable Cas9-serine recombinase fusion protein that operates 
            on DNA sequences in mammalian cells.
  JOURNAL   Nucleic Acids Res. 2016 Aug 11. pii: gkw707.
  PUBMED    27515511
REFERENCE   2  (bases 1 to 2262)
  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: "Nucleic 
            Acids Res. 2016 Aug 11. pii: gkw707."
COMMENT     Sequence Label: pHu6-gRNA-NT1
FEATURES             Location/Qualifiers
     source          1..2262
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        76..324
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     76..96
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     247..266
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        345..420
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     rep_origin      complement(513..1101)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(593..612)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1272..2063,2064..2132))
                     /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     1895..1914
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2133..2237)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacgtcgcta gctgtacaaa aaagcaggct ttaaaggaac caattcagtc gactggatcc
       61 aaggtcgggc aggaagaggg cctatttccc atgattcctt catatttgca tatacgatac
      121 aaggctgtta gagagataat tagaattaat ttgactgtaa acacaaagat attagtacaa
      181 aatacgtgac gtagaaagta ataatttctt gggtagtttg cagttttaaa attatgtttt
      241 aaaatggact atcatatgct taccgtaact tgaaagtatt tcgatttctt ggctttatat
      301 atcttgtgga aaggacgaaa caccacctct gtttgggaaa attggtttta gagctagaaa
      361 tagcaagtta aaataaggct agtccgttat caacttgaaa aagtggcacc gagtcggtgc
      421 ttttttttct cgaggtacct ctctacatat gacatgtgag caaaaggcca gcaaaaggcc
      481 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
      541 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
      601 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
      661 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
      721 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
      781 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
      841 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
      901 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
      961 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     1021 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     1081 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     1141 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     1201 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     1261 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     1321 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     1381 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     1441 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     1501 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     1561 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     1621 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     1681 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     1741 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     1801 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     1861 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     1921 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     1981 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     2041 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     2101 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     2161 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     2221 caaatagggg ttccgcgcac atttccccga aaagtgccac ct
//
