LOCUS       sequence-344929-        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"
     misc_RNA        77..152
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     rep_origin      complement(283..871)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(363..382)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1042..1833,1834..1902))
                     /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     1665..1684
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1903..2007)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        join(2114..2306,1..56)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2114..2134
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2285..2304
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
ORIGIN
        1 cttgaaagta tttcgatttc ttggctttat atatcttgtg gaaaggacga aacaccggat
       61 gatggtgagc acgtgagttt tagagctaga aatagcaagt taaaataagg ctagtccgtt
      121 atcaacttga aaaagtggca ccgagtcggt gctcaccatc atcctgcgtg ctcaccatct
      181 ttttttaagc ttgggccgct cgaggtacct ctctacatat gacatgtgag caaaaggcca
      241 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
      301 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
      361 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
      421 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
      481 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
      541 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
      601 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
      661 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
      721 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
      781 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
      841 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
      901 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
      961 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     1021 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     1081 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     1141 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagatccac gctcaccggc
     1201 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     1261 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     1321 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     1381 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     1441 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     1501 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     1561 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     1621 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     1681 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     1741 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     1801 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     1861 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     1921 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     1981 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcgc
     2041 tagctgtaca aaaaagcagg ctttaaagga accaattcag tcgactggat ccggtaccaa
     2101 ggtcgggcag gaagagggcc tatttcccat gattccttca tatttgcata tacgatacaa
     2161 ggctgttaga gagataatta gaattaattt gactgtaaac acaaagatat tagtacaaaa
     2221 tacgtgacgt agaaagtaat aatttcttgg gtagtttgca gttttaaaat tatgttttaa
     2281 aatggactat catatgctta ccgtaa
//
