LOCUS       sequence-119654-        2648 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  contains human U6 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2648)
  AUTHORS   Lonowski LA, Narimatsu Y, Riaz A, Delay CE, Yang Z, Niola F, Duda K,
            Ober EA, Clausen H, Wandall HH, Hansen SH, Bennett EP, Frodin M
  TITLE     Genome editing using FACS enrichment of nuclease-expressing cells 
            and indel detection by amplicon analysis.
  JOURNAL   Nat Protoc. 2017 Mar;12(3):581-603. doi: 10.1038/nprot.2016.165. 
            Epub 2017 Feb 16.
  PUBMED    28207001
REFERENCE   2  (bases 1 to 2648)
  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: 
            "10.1038/nprot.2016.165"; journalName: "Nat Protoc"; date: 
            "2017-03"; volume: "12"; issue: "3"; pages: "581-603"
COMMENT     Sequence Label: EPB104
FEATURES             Location/Qualifiers
     source          1..2648
                     /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"
     primer_bind     complement(791..808)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(901..1489)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(981..1000)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1660..2451,2452..2520))
                     /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     2283..2302
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2521..2625)
                     /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 aacaccggaa ttcgatatcg
      661 gtacctggag cacaagactg gcctcatggg ccttccgctc actgcccgct ttccagtcgg
      721 gaaacctgtc gtgccagctg cattaacatg gtcatagctg tttccttgcg tattgggcgc
      781 tctccgcttc ctcgctcact gactcgctgc gctcggtcgt tcgggtaaag cctggggtgc
      841 ctaatgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
      901 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
      961 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     1021 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     1081 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     1141 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     1201 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     1261 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     1321 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     1381 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     1441 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     1501 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     1561 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     1621 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     1681 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     1741 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     1801 agaaccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     1861 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     1921 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     1981 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     2041 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     2101 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     2161 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     2221 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     2281 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     2341 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     2401 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     2461 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     2521 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     2581 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     2641 agtgccac
//
