LOCUS       sequence-203528-        5574 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  W2m.2-2 plasmid. Contains UBC-TetR-P2A-LacI 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 5574)
  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 5574)
  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: pWT062a
FEATURES             Location/Qualifiers
     source          1..5574
                     /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"
     promoter        460..1670
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     1507..1526
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             2333..2389
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             3482..3502
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    3612..3746
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3649..3668)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3703..3722
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3825..4413)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3905..3924)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4584..5375,5376..5444))
                     /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     5207..5226
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5445..5549)
                     /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 ggtgacaagt gtgatcactt gttttagagc
      361 tagaaatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt
      421 cggtgctttt tttaagcttg ggccgctcga ggtagcagcg gcctccgcgc cgggttttgg
      481 cgcctcccgc gggcgccccc ctcctcacgg cgagcgctgc cacgtcagac gaagggcgca
      541 ggagcgttcc tgatccttcc gcccggacgc tcaggacagc ggcccgctgc tcataagact
      601 cggccttaga accccagtat cagcagaagg acattttagg acgggacttg ggtgactcta
      661 gggcactggt tttctttcca gagagcggaa caggcgagga aaagtagtcc cttctcggcg
      721 attctgcgga gggatctccg tggggcggtg aacgccgatg attatataag gacgcgccgg
      781 gtgtggcaca gctagttccg tcgcagccgg gatttgggtc gcggttcttg tttgtggatc
      841 gctgtgatcg tcacttggtg agttgcgggc tgctgggctg gccggggctt tcgtggccgc
      901 cgggccgctc ggtgggacgg aagcgtgtgg agagaccgcc aagggctgta gtctgggtcc
      961 gcgagcaagg ttgccctgaa ctgggggttg gggggagcgc acaaaatggc ggctgttccc
     1021 gagtcttgaa tggaagacgc ttgtaaggcg ggctgtgagg tcgttgaaac aaggtggggg
     1081 gcatggtggg cggcaagaac ccaaggtctt gaggccttcg ctaatgcggg aaagctctta
     1141 ttcgggtgag atgggctggg gcaccatctg gggaccctga cgtgaagttt gtcactgact
     1201 ggagaactcg ggtttgtcgt ctggttgcgg gggcggcagt tatgcggtgc cgttgggcag
     1261 tgcacccgta cctttgggag cgcgcgcctc gtcgtgtcgt gacgtcaccc gttctgttgg
     1321 cttataatgc agggtggggc cacctgccgg taggtgtgcg gtaggctttt ctccgtcgca
     1381 ggacgcaggg ttcgggccta gggtaggctc tcctgaatcg acaggcgccg gacctctggt
     1441 gaggggaggg ataagtgagg cgtcagtttc tttggtcggt tttatgtacc tatcttctta
     1501 agtagctgaa gctccggttt tgaactatgc gctcggggtt ggcgagtgtg ttttgtgaag
     1561 ttttttaggc accttttgaa atgtaatcat ttgggtcaat atgtaatttt cagtgttaga
     1621 ctagtaaatt gtccgctaaa ttctggccgt ttttggcttt tttgttagac aggatccccg
     1681 ggtaccggtc gccaccatgt ctcggttgga caaatctaaa gtaatcaact ctgcactgga
     1741 attgctgaac gaggtaggca tagagggcct cacaacgagg aagctggccc aaaagctggg
     1801 cgtcgaacag ccaaccctgt actggcacgt caagaataaa agggctctcc tggacgcgct
     1861 ggcaattgag atgctcgaca gacaccatac acacttttgc ccccttgaag gggaatcctg
     1921 gcaggacttc ctgcgaaaca atgccaagtc atttagatgc gctcttctgt ctcatcggga
     1981 cggtgctaag gtgcatctgg gtacaagacc cacggaaaag cagtatgaga cactggaaaa
     2041 tcaactggcc tttttgtgtc agcagggctt ctctctcgaa aacgcgcttt acgcgctgtc
     2101 agccgtgggt cattttaccc tgggctgcgt gctggaggac caggagcatc aagtggctaa
     2161 ggaggaacgg gaaaccccta ccaccgactc tatgccacct ctcttgcggc aggcaattga
     2221 gttgttcgac caccagggtg ccgagccggc cttcctgttc ggcttggagc ttatcatctg
     2281 cggcctggag aagcagctga agtgtgagag tggaagtcgt acgggaagcg gagctactaa
     2341 cttcagcctg ctgaagcagg ctggagacgt ggaggagaac cctggaccta aaccagtaac
     2401 attgtatgat gtcgcagagt atgccggtgt ctcttatcag actgtttcca gagtggtgaa
     2461 ccaggccagc catgtttctg ccaaaaccag ggaaaaagtg gaagcagcca tggcagagct
     2521 gaattacatt cccaacagag tggcacaaca actggcaggc aaacagagct tgctgattgg
     2581 agttgccacc tccagtctgg ccctgcatgc accatctcaa attgtggcag ccattaaatc
     2641 tagagctgat caactgggag cctctgtggt ggtgtcaatg gtagaaagaa gtggagttga
     2701 agcctgtaaa gctgctgtgc acaatcttct ggcacaaaga gtcagtgggc tgatcattaa
     2761 ctatccactg gatgaccagg atgccattgc tgtggaagct gcctgcacta atgttccagc
     2821 actctttctt gatgtctctg accagacacc catcaacagt attattttct cccatgaaga
     2881 tggtacaaga ctgggtgtgg agcatctggt tgcattggga caccagcaaa ttgcactgct
     2941 tgcgggccca ctcagttctg tctcagcaag gctgagactg gctggctggc ataaatatct
     3001 cactaggaat caaattcagc caatagctga aagagaaggg gactggagtg ccatgtctgg
     3061 gtttcaacaa accatgcaaa tgctgaatga gggcattgtt cccactgcaa tgctggttgc
     3121 caatgatcag atggcactgg gtgcaatgag agccattact gagtctgggc tgagagttgg
     3181 tgcagatatc tcggtagtgg gatacgacga taccgaagac agctcatgtt atatcccgcc
     3241 gttaaccacc atcaaacagg attttcgcct gctggggcaa accagcgtgg accgcttgct
     3301 gcaactctct cagggccagg cggtgaaggg caatcagctg ttgccagtct cactggtgaa
     3361 gagaaaaacc accctggcac ccaatacaca aactgcctct ccccgggcat tggctgattc
     3421 actcatgcag ctagcaagac aggtttccag actggaaagt gggcagagca gcctgaggcc
     3481 tcctaagaag aagaggaagg tttaatgatc ataatcaagc catatcacat ctgtagaggt
     3541 ttacttgctt taaaaaacct ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
     3601 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     3661 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     3721 tcaatgtatc ttatcatgtc tggatctgcc ctctctacat atgacatgtg agcaaaaggc
     3781 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     3841 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     3901 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     3961 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     4021 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     4081 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     4141 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     4201 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     4261 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     4321 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     4381 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     4441 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     4501 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     4561 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     4621 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     4681 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     4741 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     4801 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     4861 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     4921 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     4981 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     5041 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     5101 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     5161 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     5221 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     5281 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     5341 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     5401 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     5461 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     5521 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acct
//
