LOCUS       sequence-334576-        2276 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  sgRNA cloning backbone.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2276)
  AUTHORS   Choi J, Zhang T, Vu A, Ablain J, Makowski MM, Colli LM, Xu M, 
            Hennessey RC, Yin J, Rothschild H, Grawe C, Kovacs MA, Funderburk 
            KM, Brossard M, Taylor J, Pasaniuc B, Chari R, Chanock SJ, Hoggart 
            CJ, Demenais F, Barrett JH, Law MH, Iles MM, Yu K, Vermeulen M, Zon 
            LI, Brown KM
  TITLE     Massively parallel reporter assays of melanoma risk variants 
            identify MX2 as a gene promoting melanoma.
  JOURNAL   Nat Commun. 2020 Jun 1;11(1):2718. doi: 10.1038/s41467-020-16590-1.
  PUBMED    32483191
REFERENCE   2  (bases 1 to 2276)
  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: "Nat 
            Commun."; date: "2020-06-1"; volume: "11(1):2718. doi"; pages: " 
            10.1038/s41467-020-16590-1"
COMMENT     Sequence Label: pGMC00008
FEATURES             Location/Qualifiers
     source          1..2276
                     /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        271..346
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     rep_origin      complement(463..1051)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(543..562)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1222..2013,2014..2082))
                     /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     1845..1864
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2083..2187)
                     /gene="bla"
                     /label=AmpR promoter
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 gagacgatta atgcgtctcg gttttagagc tagaaatagc aagttaaaat
      301 aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt tttgaattct
      361 cgacctcgag acaaatggct cgaggtacct ctctacatat gacatgtgag caaaaggcca
      421 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
      481 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
      541 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
      601 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
      661 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
      721 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
      781 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
      841 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
      901 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
      961 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     1021 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     1081 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     1141 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     1201 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     1261 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     1321 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     1381 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     1441 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     1501 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     1561 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     1621 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     1681 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     1741 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     1801 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     1861 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     1921 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     1981 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     2041 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     2101 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     2161 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcgc
     2221 tagctgtaca aaaaagcagg ctttaaagga accaattcag tcgactggat cccacc
//
