LOCUS       sequence-355981-        4876 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PB-10X IRF-minCMV-BFP-Triplex-crAAVS1_8-SV40pA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4876)
  AUTHORS   Kempton HR, Love KS, Guo LY, Qi LS
  TITLE     Scalable biological signal recording in mammalian cells using Cas12a
            base editors.
  JOURNAL   Nat Chem Biol. 2022 May 30. pii: 10.1038/s41589-022-01034-2. doi: 
            10.1038/s41589-022-01034-2.
  PUBMED    35637351
REFERENCE   2  (bases 1 to 4876)
  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: "Nat Chem 
            Biol. 2022 May 30. pii: 10.1038/s41589-022-01034-2. doi: 
            10.1038/s41589-022-01034-2."
COMMENT     Sequence Label: pSLQ11465
FEATURES             Location/Qualifiers
     source          1..4876
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   5..39
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        503..540
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     537..561
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             596..1297
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     primer_bind     complement(1603..1622)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1657..1676
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   1910..1972
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(2019..2039)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2019..2037)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2058..2074)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2058..2074)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2071..2093)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2082..2098
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(2106..2112,2113..2130,2131..2136))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2151..2172
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2289..2306)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2460..3048)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2540..2559)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3219..4010,4011..4079))
                     /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     3842..3861
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4080..4184)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4210..4665)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4347..4368)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4559..4578
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4792..4814
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4806..4823
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4807..4823
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4833..4852
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4833..4851
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ttaaccctag aaagatagtc tgcgtaaaat tgacgcatgc attcttgaaa tattgctctc
       61 tctttctaaa tagcgcgaat ccgtcgctgt gcatttagga catctcagtc gccgcttgga
      121 gctcccgtga ggcgtgcttg tcaatgcggt aagtgtcact gattttgaac tataacgacc
      181 gcgtgagtca aaatgacgca tgattatctt ttacgtgact tttaagattt aactcatacg
      241 ataattatat tgttatttca tgttctactt acgtgataac ttattatata tatattttct
      301 tgttatagat atcaactaga atgctagcta gtttcacttt ccctagtttc actttcccta
      361 gtttcacttt ccctagtttc actttcccta gtttcacttt cccgtcgact agtttcactt
      421 tccctagttt cactttccct agtttcactt tccctagttt cactttccct agtttcactt
      481 tccccccggg ctcgacatat gcgtaggcgt gtacggtggg aggtctatat aagcagagct
      541 cgtttagtga accgtcagat cctagtctac tacccagaaa gcttgaattc ccaccatgag
      601 cgagctgatt aaggagaaca tgcacatgaa gctgtacatg gagggcaccg tggacaacca
      661 tcacttcaag tgcacatccg agggcgaagg caagccctac gagggcaccc agaccatgag
      721 aatcaaggtg gtcgagggcg gccctctccc cttcgccttc gacatcctgg ctactagctt
      781 cctctacggc agcaagacct tcatcaacca cacccagggc atccccgact tcttcaagca
      841 gtccttccct gagggcttca catgggagag agtcaccaca tacgaagacg ggggcgtgct
      901 gaccgctacc caggacacca gcctccagga cggctgcctc atctacaacg tcaagatcag
      961 aggggtgaac ttcacatcca acggccctgt gatgcagaag aaaacactcg gctgggaggc
     1021 cttcaccgag actctgtatc ctgctgacgg cggcctggaa ggcagaaacg acatggccct
     1081 gaagctcgtg ggcgggagcc atctgatcgc aaacatcaag accacatata gatccaagaa
     1141 acccgctaag aacctcaaga tgcctggcgt ctactatgtg gactacagac tggaaagaat
     1201 caaggaggcc aacaacgaga cctacgtcga gcagcacgag gtggcagtgg ccagatactg
     1261 cgacctccct agcaaactgg ggcacaagct taattaaacg cgtgattcgt cagtagggtt
     1321 gtaaaggttt ttcttttcct gagaaaacaa ccttttgttt tctcaggttt tgctttttgg
     1381 cctttcccta gctttaaaaa aaaaaaagca aaactcaccg aggcagttcc ataggatggc
     1441 aagatcctgg tattggtctg cgaggatcct ttctttcttt gaatttctac taagtgtaga
     1501 tatttcccag catgccctgc gaggaatttc tactaagtgt agatcgagtt ttctttttgt
     1561 ttctcgagtt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     1621 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     1681 tatcttatca tgtctggatc aactggataa ctcaagctaa ccgactagtc tagttaaaag
     1741 ttttgttact ttatagaaga aattttgagt ttttgttttt ttttaataaa taaataaaca
     1801 taaataaatt gtttgttgaa tttattatta gtatgtaagt gtaaatataa taaaacttaa
     1861 tatctattca aattaataaa taaacctcga tatacagacc gataaaacac atgcgtcaat
     1921 tttacgcatg attatcttta acgtacgtca caatatgatt atctttctag ggttaatcta
     1981 gtgtttaaac ggccgcttaa ttaatccagc ttttgttccc tttagtgagg gttaattgcg
     2041 cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     2101 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     2161 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     2221 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     2281 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     2341 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     2401 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     2461 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     2521 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     2581 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     2641 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     2701 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     2761 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     2821 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     2881 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc
     2941 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     3001 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     3061 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     3121 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     3181 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     3241 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     3301 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     3361 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     3421 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     3481 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     3541 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     3601 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     3661 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     3721 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     3781 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     3841 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     3901 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     3961 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     4021 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     4081 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     4141 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     4201 gtgccaccta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     4261 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
     4321 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     4381 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     4441 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     4501 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     4561 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     4621 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat tcgccattca
     4681 ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg
     4741 cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac
     4801 gacgttgtaa aacgacggcc agtgagcgcg cgtaatacga ctcactatag ggcgaattgg
     4861 ggcgcgccat tctaga
//
