LOCUS       sequence-361639-        4891 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PB-STAT6RE-minCMV-BFP-Triplex-crAAVS1_6-SV40pA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4891)
  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 4891)
  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: pSLQ11474
FEATURES             Location/Qualifiers
     source          1..4891
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   22..56
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        538..575
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     572..596
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             631..1332
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     primer_bind     complement(1635..1654)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1689..1708
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   1942..2004
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(2051..2071)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2051..2069)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2090..2106)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2090..2106)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2103..2125)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2114..2130
                     /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(2138..2144,2145..2162,2163..2168))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2183..2204
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2321..2338)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2492..3080)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2572..2591)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3251..4042,4043..4111))
                     /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     3874..3893
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4112..4216)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4242..4697)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4379..4400)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4591..4610
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4824..4846
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4838..4855
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4839..4855
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4865..4884
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4865..4883
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gggcgcgcca ttctagatta accctagaaa gatagtctgc gtaaaattga cgcatgcatt
       61 cttgaaatat tgctctctct ttctaaatag cgcgaatccg tcgctgtgca tttaggacat
      121 ctcagtcgcc gcttggagct cccgtgaggc gtgcttgtca atgcggtaag tgtcactgat
      181 tttgaactat aacgaccgcg tgagtcaaaa tgacgcatga ttatctttta cgtgactttt
      241 aagatttaac tcatacgata attatattgt tatttcatgt tctacttacg tgataactta
      301 ttatatatat attttcttgt tatagatatc aactagaatg ctagccgctg ttgctcaatc
      361 gacttcccaa gaacagcgct gttgctcaat cgacttccca agaacagcag ggatctggcc
      421 tcggcggcca agctccgctg ttgctcaatc gacttcccaa gaacagcgct gttgctcaat
      481 cgacttccca agaacagcag ggatctggcc tcggcggccc ccgggctcga catatgcgta
      541 ggcgtgtacg gtgggaggtc tatataagca gagctcgttt agtgaaccgt cagatcctag
      601 tctactaccc agaaagcttg aattcccacc atgagcgagc tgattaagga gaacatgcac
      661 atgaagctgt acatggaggg caccgtggac aaccatcact tcaagtgcac atccgagggc
      721 gaaggcaagc cctacgaggg cacccagacc atgagaatca aggtggtcga gggcggccct
      781 ctccccttcg ccttcgacat cctggctact agcttcctct acggcagcaa gaccttcatc
      841 aaccacaccc agggcatccc cgacttcttc aagcagtcct tccctgaggg cttcacatgg
      901 gagagagtca ccacatacga agacgggggc gtgctgaccg ctacccagga caccagcctc
      961 caggacggct gcctcatcta caacgtcaag atcagagggg tgaacttcac atccaacggc
     1021 cctgtgatgc agaagaaaac actcggctgg gaggccttca ccgagactct gtatcctgct
     1081 gacggcggcc tggaaggcag aaacgacatg gccctgaagc tcgtgggcgg gagccatctg
     1141 atcgcaaaca tcaagaccac atatagatcc aagaaacccg ctaagaacct caagatgcct
     1201 ggcgtctact atgtggacta cagactggaa agaatcaagg aggccaacaa cgagacctac
     1261 gtcgagcagc acgaggtggc agtggccaga tactgcgacc tccctagcaa actggggcac
     1321 aagcttaatt aaacgcgtga ttcgtcagta gggttgtaaa ggtttttctt ttcctgagaa
     1381 aacaaccttt tgttttctca ggttttgctt tttggccttt ccctagcttt aaaaaaaaaa
     1441 aagcaaaact caccgaggca gttccatagg atggcaagat cctggtattg gtctgcgagg
     1501 atcctttctt tctttgaatt tctactaagt gtagatatgg cttctaggtt gcggacaatt
     1561 tctactaagt gtagatcgag ttttcttttt gtttctcgag ttgtttattg cagcttataa
     1621 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     1681 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga tcaactggat
     1741 aactcaagct aaccgactag tctagttaaa agttttgtta ctttatagaa gaaattttga
     1801 gtttttgttt ttttttaata aataaataaa cataaataaa ttgtttgttg aatttattat
     1861 tagtatgtaa gtgtaaatat aataaaactt aatatctatt caaattaata aataaacctc
     1921 gatatacaga ccgataaaac acatgcgtca attttacgca tgattatctt taacgtacgt
     1981 cacaatatga ttatctttct agggttaatc tagtgtttaa acggccgctt aattaatcca
     2041 gcttttgttc cctttagtga gggttaattg cgcgcttggc gtaatcatgg tcatagctgt
     2101 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     2161 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     2221 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     2281 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     2341 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     2401 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     2461 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     2521 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     2581 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     2641 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     2701 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     2761 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     2821 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     2881 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     2941 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     3001 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     3061 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     3121 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     3181 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     3241 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     3301 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     3361 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     3421 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     3481 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     3541 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     3601 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     3661 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     3721 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     3781 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     3841 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     3901 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     3961 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     4021 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     4081 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     4141 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     4201 aaataggggt tccgcgcaca tttccccgaa aagtgccacc taaattgtaa gcgttaatat
     4261 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     4321 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     4381 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     4441 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     4501 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     4561 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     4621 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     4681 gccgctacag ggcgcgtccc attcgccatt caggctgcgc aactgttggg aagggcgatc
     4741 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
     4801 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgagcg
     4861 cgcgtaatac gactcactat agggcgaatt g
//
