LOCUS       sequence-360581-        5020 bp    DNA     circular SYN 14-APR-2025
DEFINITION  PB-TRE3G-BFP-Triplex-crGFP*ABE3-SV40pA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5020)
  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 5020)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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: pSLQ11350
FEATURES             Location/Qualifiers
     source          1..5020
                     /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)"
     protein_bind    358..376
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    394..412
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    430..448
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    466..484
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    502..520
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    538..556
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    574..592
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     637..661
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             740..1441
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     primer_bind     complement(1747..1766)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1801..1820
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   2054..2116
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(2163..2183)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2163..2181)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2202..2218)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2202..2218)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2215..2237)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2226..2242
                     /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(2250..2256,2257..2274,2275..2280))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2295..2316
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2433..2450)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2604..3192)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2684..2703)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3363..4154,4155..4223))
                     /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     3986..4005
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4224..4328)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4354..4809)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4491..4512)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4703..4722
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4936..4958
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4950..4967
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4951..4967
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4977..4996
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4977..4995
                     /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 atgctagcgc tgatatctcc accccacagt ggtttactcc
      361 ctatcagtga tagagaacgt atgaagagtt tactccctat cagtgataga gaacgtatgc
      421 agactttact ccctatcagt gatagagaac gtataaggag tttactccct atcagtgata
      481 gagaacgtat gaccagttta ctccctatca gtgatagaga acgtatctac agtttactcc
      541 ctatcagtga tagagaacgt atatccagtt tactccctat cagtgataga gaacgtataa
      601 gctttaggcg tgtacggtgg gcgcctataa aagcagagct cgtttagtga accgtcagat
      661 cgcctggagc aattccacaa cacttttgtc ttataccaac tttccgtacc acttcctacc
      721 ctcgtaaaga attcccacca tgagcgagct gattaaggag aacatgcaca tgaagctgta
      781 catggagggc accgtggaca accatcactt caagtgcaca tccgagggcg aaggcaagcc
      841 ctacgagggc acccagacca tgagaatcaa ggtggtcgag ggcggccctc tccccttcgc
      901 cttcgacatc ctggctacta gcttcctcta cggcagcaag accttcatca accacaccca
      961 gggcatcccc gacttcttca agcagtcctt ccctgagggc ttcacatggg agagagtcac
     1021 cacatacgaa gacgggggcg tgctgaccgc tacccaggac accagcctcc aggacggctg
     1081 cctcatctac aacgtcaaga tcagaggggt gaacttcaca tccaacggcc ctgtgatgca
     1141 gaagaaaaca ctcggctggg aggccttcac cgagactctg tatcctgctg acggcggcct
     1201 ggaaggcaga aacgacatgg ccctgaagct cgtgggcggg agccatctga tcgcaaacat
     1261 caagaccaca tatagatcca agaaacccgc taagaacctc aagatgcctg gcgtctacta
     1321 tgtggactac agactggaaa gaatcaagga ggccaacaac gagacctacg tcgagcagca
     1381 cgaggtggca gtggccagat actgcgacct ccctagcaaa ctggggcaca agcttaatta
     1441 aacgcgtgat tcgtcagtag ggttgtaaag gtttttcttt tcctgagaaa acaacctttt
     1501 gttttctcag gttttgcttt ttggcctttc cctagcttta aaaaaaaaaa agcaaaactc
     1561 accgaggcag ttccatagga tggcaagatc ctggtattgg tctgcgagga tcctttcttt
     1621 ctttgaattt ctactaagtg tagatactcg attcagttaa caagggtaaa tttctactaa
     1681 gtgtagatcg agttttcttt ttgtttctcg agttgtttat tgcagcttat aatggttaca
     1741 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     1801 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatcaactgg ataactcaag
     1861 ctaaccgact agtctagtta aaagttttgt tactttatag aagaaatttt gagtttttgt
     1921 ttttttttaa taaataaata aacataaata aattgtttgt tgaatttatt attagtatgt
     1981 aagtgtaaat ataataaaac ttaatatcta ttcaaattaa taaataaacc tcgatataca
     2041 gaccgataaa acacatgcgt caattttacg catgattatc tttaacgtac gtcacaatat
     2101 gattatcttt ctagggttaa tctagtgttt aaacggccgc ttaattaatc cagcttttgt
     2161 tccctttagt gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg
     2221 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     2281 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     2341 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     2401 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     2461 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     2521 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     2581 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     2641 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     2701 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     2761 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     2821 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     2881 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     2941 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     3001 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     3061 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     3121 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     3181 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     3241 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     3301 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     3361 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     3421 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     3481 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     3541 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     3601 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     3661 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     3721 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     3781 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     3841 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     3901 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     3961 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     4021 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     4081 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     4141 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     4201 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     4261 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     4321 gttccgcgca catttccccg aaaagtgcca cctaaattgt aagcgttaat attttgttaa
     4381 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
     4441 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
     4501 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     4561 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
     4621 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     4681 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     4741 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     4801 agggcgcgtc ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
     4861 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg
     4921 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgag cgcgcgtaat
     4981 acgactcact atagggcgaa ttggggcgcg ccattctaga
//
