LOCUS       sequence-340873-        2306 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressing a single clone of a 16 variant library of pegRNAs that 
            inserts CAGGA to TAPE-1..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2306)
  AUTHORS   Choi J, Chen W, Minkina A, Chardon FM, Suiter CC, Regalado SG, 
            Domcke S, Hamazaki N, Lee C, Martin B, Daza RM, Shendure J
  TITLE     A time-resolved, multi-symbol molecular recorder via sequential 
            genome editing.
  JOURNAL   Nature. 2022 Jul 6. pii: 10.1038/s41586-022-04922-8. doi: 
            10.1038/s41586-022-04922-8.
  PUBMED    35794474
REFERENCE   2  (bases 1 to 2306)
  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: "Nature. 
            2022 Jul 6. pii: 10.1038/s41586-022-04922-8. doi: 
            10.1038/s41586-022-04922-8."
COMMENT     Sequence Label: pU6-CApegTAPE1
FEATURES             Location/Qualifiers
     source          1..2306
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        82..330
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     82..102
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     253..272
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        351..426
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     rep_origin      complement(557..1145)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(637..656)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1316..2107,2108..2176))
                     /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     1939..1958
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2177..2281)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacgtcgcta gctgtacaaa aaagcaggct ttaaaggaac caattcagtc gactggatcc
       61 ggtaccaagg tcgggcagga agagggccta tttcccatga ttccttcata tttgcatata
      121 cgatacaagg ctgttagaga gataattaga attaatttga ctgtaaacac aaagatatta
      181 gtacaaaata cgtgacgtag aaagtaataa tttcttgggt agtttgcagt tttaaaatta
      241 tgttttaaaa tggactatca tatgcttacc gtaacttgaa agtatttcga tttcttggct
      301 ttatatatct tgtggaaagg acgaaacacc ggatgatggt gagcacgtga gttttagagc
      361 tagaaatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt
      421 cggtgctcac catcatccnn cgtgctcacc atcttttttt aagcttgggc cgctcgaggt
      481 acctctctac atatgacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
      541 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
      601 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
      661 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
      721 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
      781 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
      841 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
      901 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
      961 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     1021 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     1081 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     1141 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     1201 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     1261 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     1321 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     1381 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     1441 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     1501 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     1561 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     1621 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     1681 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     1741 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     1801 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     1861 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     1921 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     1981 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     2041 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     2101 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     2161 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     2221 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     2281 gcacatttcc ccgaaaagtg ccacct
//
