LOCUS       sequence-360580-        4841 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PB-NFKB-BFP-Triplex-crAAVS1_4-SV40pA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4841)
  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 4841)
  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: pSLQ11453
FEATURES             Location/Qualifiers
     source          1..4841
                     /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)"
     primer_bind     466..490
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             564..1265
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     primer_bind     complement(1568..1587)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1622..1641
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   1875..1937
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(1984..2004)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1984..2002)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2023..2039)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2023..2039)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2036..2058)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2047..2063
                     /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(2071..2077,2078..2095,2096..2101))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2116..2137
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2254..2271)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2425..3013)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2505..2524)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3184..3975,3976..4044))
                     /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     3807..3826
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4045..4149)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4175..4630)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4312..4333)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4524..4543
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4757..4779
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4771..4788
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4772..4788
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4798..4817
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4798..4816
                     /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 atgctagccg ctctagacca gatccgggga ctttccgctt
      361 ggggactttc cgctggggac tttccgctgg ggactttccg ctggggactt tccgaattca
      421 gcttgaggta cctaggcgtg tacggtggga ggcctatata agcagagctc gtttagtgaa
      481 ccgtcagatc gcctggagac gccatccacg ctgttttgac ctccatagaa gacaccggga
      541 ccgatccagc ctgaattccc accatgagcg agctgattaa ggagaacatg cacatgaagc
      601 tgtacatgga gggcaccgtg gacaaccatc acttcaagtg cacatccgag ggcgaaggca
      661 agccctacga gggcacccag accatgagaa tcaaggtggt cgagggcggc cctctcccct
      721 tcgccttcga catcctggct actagcttcc tctacggcag caagaccttc atcaaccaca
      781 cccagggcat ccccgacttc ttcaagcagt ccttccctga gggcttcaca tgggagagag
      841 tcaccacata cgaagacggg ggcgtgctga ccgctaccca ggacaccagc ctccaggacg
      901 gctgcctcat ctacaacgtc aagatcagag gggtgaactt cacatccaac ggccctgtga
      961 tgcagaagaa aacactcggc tgggaggcct tcaccgagac tctgtatcct gctgacggcg
     1021 gcctggaagg cagaaacgac atggccctga agctcgtggg cgggagccat ctgatcgcaa
     1081 acatcaagac cacatataga tccaagaaac ccgctaagaa cctcaagatg cctggcgtct
     1141 actatgtgga ctacagactg gaaagaatca aggaggccaa caacgagacc tacgtcgagc
     1201 agcacgaggt ggcagtggcc agatactgcg acctccctag caaactgggg cacaagctta
     1261 attaaacgcg tgattcgtca gtagggttgt aaaggttttt cttttcctga gaaaacaacc
     1321 ttttgttttc tcaggttttg ctttttggcc tttccctagc tttaaaaaaa aaaaagcaaa
     1381 actcaccgag gcagttccat aggatggcaa gatcctggta ttggtctgcg aggatccttt
     1441 ctttctttga atttctacta agtgtagatt cttctgtgca ctacttgtca atttctacta
     1501 agtgtagatc gagttttctt tttgtttctc gagttgttta ttgcagctta taatggttac
     1561 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     1621 tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatcaactg gataactcaa
     1681 gctaaccgac tagtctagtt aaaagttttg ttactttata gaagaaattt tgagtttttg
     1741 ttttttttta ataaataaat aaacataaat aaattgtttg ttgaatttat tattagtatg
     1801 taagtgtaaa tataataaaa cttaatatct attcaaatta ataaataaac ctcgatatac
     1861 agaccgataa aacacatgcg tcaattttac gcatgattat ctttaacgta cgtcacaata
     1921 tgattatctt tctagggtta atctagtgtt taaacggccg cttaattaat ccagcttttg
     1981 ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt
     2041 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     2101 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     2161 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     2221 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     2281 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     2341 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     2401 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     2461 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     2521 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     2581 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     2641 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     2701 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     2761 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     2821 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     2881 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     2941 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     3001 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     3061 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     3121 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     3181 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     3241 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     3301 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     3361 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     3421 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     3481 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     3541 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     3601 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     3661 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     3721 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     3781 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     3841 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     3901 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     3961 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     4021 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     4081 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     4141 ggttccgcgc acatttcccc gaaaagtgcc acctaaattg taagcgttaa tattttgtta
     4201 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     4261 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     4321 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     4381 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     4441 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     4501 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     4561 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     4621 cagggcgcgt cccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
     4681 gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg
     4741 gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga gcgcgcgtaa
     4801 tacgactcac tatagggcga attggggcgc gccattctag a
//
