LOCUS       sequence-203530-        2216 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  W2m.2-4 plasmid. Contains H1-TetR-sgRNA B and is ppart of the 
            CAMERA2m system for cellular recording in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2216)
  AUTHORS   Tang W, Liu DR
  TITLE     Rewritable multi-event analog recording in bacterial and mammalian 
            cells.
  JOURNAL   Science. 2018 Feb 15. pii: science.aap8992. doi: 
            10.1126/science.aap8992.
  PUBMED    29449507
REFERENCE   2  (bases 1 to 2216)
  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: "Science. 
            2018 Feb 15. pii: science.aap8992. doi: 10.1126/science.aap8992."
COMMENT     Sequence Label: pWT062d
FEATURES             Location/Qualifiers
     source          1..2216
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     180..199
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     protein_bind    244..262
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_RNA        288..363
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     rep_origin      complement(467..1055)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(547..566)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1226..2017,2018..2086))
                     /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     1849..1868
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2087..2191)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gcgaacgctc ctcaccatac gaaagacctt gaatcctatg cttcgaacgc tcacgtcatc
       61 aacccgctcc aaggaatcgc gggcccagtg tcactaggcg ggaacaccca gcgcgcgtgc
      121 gccctggcag gaagatggct gtgagggaca ggggagtggc gccctgcaat atttgcatgt
      181 cgctatgtgt tctgggaaat caccataaac gtgaaatgtc tttggatttg ggaatcttat
      241 aagtccctat cagtgataga gatcccacag gacggtcacc tttgggggtt ttagagctag
      301 aaatagcaag ttaaaataag gctagtccgt tatcaacttg aaaaagtggc accgagtcgg
      361 tgcttttttt aagcttgggc cgctcgaggt acctctctac atatgacatg tgagcaaaag
      421 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
      481 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
      541 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
      601 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
      661 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
      721 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
      781 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
      841 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
      901 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
      961 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     1021 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     1081 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     1141 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     1201 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     1261 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     1321 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     1381 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     1441 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     1501 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     1561 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     1621 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     1681 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     1741 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     1801 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     1861 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     1921 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     1981 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     2041 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     2101 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     2161 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacct
//
