LOCUS       sequence-361633-        4875 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 4875)
  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 4875)
  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..4875
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   24..58
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        522..559
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     556..580
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             615..1316
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     primer_bind     complement(1621..1640)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1675..1694
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   1928..1990
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(2037..2057)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2037..2055)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2076..2092)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2076..2092)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2089..2111)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2100..2116
                     /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(2124..2130,2131..2148,2149..2154))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2169..2190
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2307..2324)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2478..3066)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2558..2577)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3237..4028,4029..4097))
                     /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     3860..3879
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4098..4202)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4228..4683)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4365..4386)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4577..4596
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4810..4832
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4824..4841
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4825..4841
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4851..4870
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4851..4869
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 tggggcgcgc cattctagat taaccctaga aagatagtct gcgtaaaatt gacgcatgca
       61 ttcttgaaat attgctctct ctttctaaat agcgcgaatc cgtcgctgtg catttaggac
      121 atctcagtcg ccgcttggag ctcccgtgag gcgtgcttgt caatgcggta agtgtcactg
      181 attttgaact ataacgaccg cgtgagtcaa aatgacgcat gattatcttt tacgtgactt
      241 ttaagattta actcatacga taattatatt gttatttcat gttctactta cgtgataact
      301 tattatatat atattttctt gttatagata tcaactagaa tgctagctag tttcactttc
      361 cctagtttca ctttccctag tttcactttc cctagtttca ctttccctag tttcactttc
      421 ccgtcgacta gtttcacttt ccctagtttc actttcccta gtttcacttt ccctagtttc
      481 actttcccta gtttcacttt ccccccgggc tcgacatatg cgtaggcgtg tacggtggga
      541 ggtctatata agcagagctc gtttagtgaa ccgtcagatc ctagtctact acccagaaag
      601 cttgaattcc caccatgagc gagctgatta aggagaacat gcacatgaag ctgtacatgg
      661 agggcaccgt ggacaaccat cacttcaagt gcacatccga gggcgaaggc aagccctacg
      721 agggcaccca gaccatgaga atcaaggtgg tcgagggcgg ccctctcccc ttcgccttcg
      781 acatcctggc tactagcttc ctctacggca gcaagacctt catcaaccac acccagggca
      841 tccccgactt cttcaagcag tccttccctg agggcttcac atgggagaga gtcaccacat
      901 acgaagacgg gggcgtgctg accgctaccc aggacaccag cctccaggac ggctgcctca
      961 tctacaacgt caagatcaga ggggtgaact tcacatccaa cggccctgtg atgcagaaga
     1021 aaacactcgg ctgggaggcc ttcaccgaga ctctgtatcc tgctgacggc ggcctggaag
     1081 gcagaaacga catggccctg aagctcgtgg gcgggagcca tctgatcgca aacatcaaga
     1141 ccacatatag atccaagaaa cccgctaaga acctcaagat gcctggcgtc tactatgtgg
     1201 actacagact ggaaagaatc aaggaggcca acaacgagac ctacgtcgag cagcacgagg
     1261 tggcagtggc cagatactgc gacctcccta gcaaactggg gcacaagctt aattaaacgc
     1321 gtgattcgtc agtagggttg taaaggtttt tcttttcctg agaaaacaac cttttgtttt
     1381 ctcaggtttt gctttttggc ctttccctag ctttaaaaaa aaaaaagcaa aactcaccga
     1441 ggcagttcca taggatggca agatcctggt attggtctgc gaggatcctt ctttctttga
     1501 atttctacta agtgtagata tttcccagca tgccctgcga ggaatttcta ctaagtgtag
     1561 atcgagtttt ctttttgttt ctcgagttgt ttattgcagc ttataatggt tacaaataaa
     1621 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     1681 tgtccaaact catcaatgta tcttatcatg tctggatcaa ctggataact caagctaacc
     1741 gactagtcta gttaaaagtt ttgttacttt atagaagaaa ttttgagttt ttgttttttt
     1801 ttaataaata aataaacata aataaattgt ttgttgaatt tattattagt atgtaagtgt
     1861 aaatataata aaacttaata tctattcaaa ttaataaata aacctcgata tacagaccga
     1921 taaaacacat gcgtcaattt tacgcatgat tatctttaac gtacgtcaca atatgattat
     1981 ctttctaggg ttaatctagt gtttaaacgg ccgcttaatt aatccagctt ttgttccctt
     2041 tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     2101 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     2161 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     2221 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     2281 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     2341 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     2401 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     2461 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     2521 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     2581 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     2641 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     2701 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     2761 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     2821 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     2881 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     2941 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     3001 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     3061 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     3121 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     3181 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     3241 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     3301 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     3361 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     3421 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     3481 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     3541 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     3601 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     3661 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     3721 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     3781 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     3841 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     3901 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     3961 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     4021 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     4081 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     4141 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     4201 cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt taatattttg ttaaaattcg
     4261 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
     4321 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
     4381 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     4441 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
     4501 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
     4561 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
     4621 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
     4681 cgtcccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt
     4741 cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc
     4801 cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgagcgcgcg taatacgact
     4861 cactataggg cgaat
//
