LOCUS       sequence-207311-        5576 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 5576)
  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 5576)
  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..5576
                     /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..1671
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     1508..1527
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             2334..2390
                     /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             3483..3503
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    3614..3748
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3651..3670)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3705..3724
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3827..4415)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3907..3926)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4586..5377,5378..5446))
                     /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     5209..5228
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5447..5551)
                     /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 gcgagcgtcc 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 agtagcgggc tgctgggctg gccggggctt tcgtggccgc
      901 cgggccgctc ggtgggacgg aagcgtgtgg agagaccgcc aagggctgta gtctgggtcc
      961 gcgagcaagg ttgccctgaa ctgggggttg gggggagcgc agcaaaatgg cggctgttcc
     1021 cgagtcttga atggaagacg cttgtgaggc gggctgtgag gtcgttgaaa caaggtgggg
     1081 ggcatggtgg gcggcaagaa cccaaggtct tgaggccttc gctaatgcgg gaaagctctt
     1141 attcgggtga gatgggctgg ggcaccatct ggggaccctg acgtgaagtt tgtcactgac
     1201 tggagaactc ggtttgtcgt ctgttgcggg ggcggcagtt atggcggtgc cgttgggcag
     1261 tgcacccgta cctttgggag cgcgcgccct cgtcgtgtcg tgacgtcacc cgttctgttg
     1321 gcttataatg cagggtgggg ccacctgccg gtaggtgtgc ggtaggcttt tctccgtcgc
     1381 aggacgcagg gttcgggcct agggtaggct ctcctgaatc gacaggcgcc ggacctctgg
     1441 tgaggggagg gataagtgag gcgtcagttt ctttggtcgg ttttatgtac ctatcttctt
     1501 aagtagctga agctccggtt ttgaactatg cgctcggggt tggcgagtgt gttttgtgaa
     1561 gttttttagg caccttttga aatgtaatca tttgggtcaa tatgtaattt tcagtgttag
     1621 actagtaaat tgtccgctaa attctggccg tttttggctt ttttgttaga caggatcccc
     1681 gggtaccggt cgccaccatg tctcggttgg acaaatctaa agtaatcaac tctgcactgg
     1741 aattgctgaa cgaggtaggc atagagggcc tcacaacgag gaagctggcc caaaagctgg
     1801 gcgtcgaaca gccaaccctg tactggcacg tcaagaataa aagggctctc ctggacgcgc
     1861 tggcaattga gatgctcgac agacaccata cacacttttg cccccttgaa ggggaatcct
     1921 ggcaggactt cctgcgaaac aatgccaagt catttagatg cgctcttctg tctcatcggg
     1981 acggtgctaa ggtgcatctg ggtacaagac ccacggaaaa gcagtatgag acactggaaa
     2041 atcaactggc ctttttgtgt cagcagggct tctctctcga aaacgcgctt tacgcgctgt
     2101 cagccgtggg tcattttacc ctgggctgcg tgctggagga ccaggagcat caagtggcta
     2161 aggaggaacg ggaaacccct accaccgact ctatgccacc tctcttgcgg caggcaattg
     2221 agttgttcga ccaccagggt gccgagccgg ccttcctgtt cggcttggag cttatcatct
     2281 gcggcctgga gaagcagctg aagtgtgaga gtggaagtcg tacgggaagc ggagctacta
     2341 acttcagcct gctgaagcag gctggagacg tggaggagaa ccctggacct aaaccagtaa
     2401 cattgtatga tgtcgcagag tatgccggtg tctcttatca gactgtttcc agagtggtga
     2461 accaggccag ccatgtttct gccaaaacca gggaaaaagt ggaagcagcc atggcagagc
     2521 tgaattacat tcccaacaga gtggcacaac aactggcagg caaacagagc ttgctgattg
     2581 gagttgccac ctccagtctg gccctgcatg caccatctca aattgtggca gccattaaat
     2641 ctagagctga tcaactggga gcctctgtgg tggtgtcaat ggtagaaaga agtggagttg
     2701 aagcctgtaa agctgctgtg cacaatcttc tggcacaaag agtcagtggg ctgatcatta
     2761 actatccact ggatgaccag gatgccattg ctgtggaagc tgcctgcact aatgttccag
     2821 cactctttct tgatgtctct gaccagacac ccatcaacag tattattttc tcccatgaag
     2881 atggtacaag actgggtgtg gagcatctgg ttgcattggg acaccagcaa attgcactgc
     2941 ttgcgggccc actcagttct gtctcagcaa ggctgagact ggctggctgg cataaatatc
     3001 tcactaggaa tcaaattcag ccaatagctg aaagagaagg ggactggagt gccatgtctg
     3061 ggtttcaaca aaccatgcaa atgctgaatg agggcattgt tcccactgca atgctggttg
     3121 ccaatgatca gatggcactg ggtgcaatga gagccattac tgagtctggg ctgagagttg
     3181 gtgcagatat ctcggtagtg ggatacgacg ataccgaaga cagctcatgt tatatcccgc
     3241 cgttaaccac catcaaacag gattttcgcc tgctggggca aaccagcgtg gaccgcttgc
     3301 tgcaactctc tcagggccag gcggtgaagg gcaatcagct gttgccagtc tcactggtga
     3361 agagaaaaac caccctggca cccaatacac aaactgcctc tccccgggca ttggctgatt
     3421 cactcatgca gctagcaaga caggtttcca gactggaaag tgggcagagc agcctgaggc
     3481 ctcctaagaa gaagaggaag gtttaatgat cataatcaag ccatatcaca tctgtagagg
     3541 tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc
     3601 aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     3661 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     3721 catcaatgta tcttatcatg tctggatctg ccctctctac atatgacatg tgagcaaaag
     3781 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     3841 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     3901 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     3961 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     4021 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     4081 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     4141 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     4201 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     4261 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     4321 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     4381 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     4441 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     4501 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     4561 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     4621 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     4681 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     4741 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     4801 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     4861 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     4921 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     4981 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     5041 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     5101 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     5161 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     5221 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     5281 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     5341 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     5401 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     5461 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     5521 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacct
//
