LOCUS       sequence-339407-        8076 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses BEACON2 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8076)
  AUTHORS   Wang X, Ding C, Yu W, Wang Y, He S, Yang B, Xiong YC, Wei J, Li J, 
            Liang J, Lu Z, Zhu W, Wu J, Zhou Z, Huang X, Liu Z, Yang L, Chen J
  TITLE     Cas12a Base Editors Induce Efficient and Specific Editing with Low 
            DNA Damage Response.
  JOURNAL   Cell Rep. 2020 Jun 2;31(9):107723. doi: 
            10.1016/j.celrep.2020.107723.
  PUBMED    32492431
REFERENCE   2  (bases 1 to 8076)
  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: "Cell Rep.";
            date: "2020-06-2"; volume: "31(9):107723. doi"; pages: " 
            10.1016/j.celrep.2020.107723"
COMMENT     Sequence Label: pCMV-BEACON2
FEATURES             Location/Qualifiers
     source          1..8076
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        132..335
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     286..306
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     377..396
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        377..395
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             412..432
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1078..4761
                     /codon_start=1
                     /product="CRISPR-associated protein from Lachnospiraceae 
                     bacterium ND2006 (Zetsche et al., 2015)"
                     /label=LbCpf1
                     /translation="MSKLEKFTNCYSLSKTLRFKAIPVGKTQENIDNKRLLVEDEKRAE
                     DYKGVKKLLDRYYLSFINDVLHSIKLKNLNNYISLFRKKTRTEKENKELENLEINLRKE
                     IAKAFKGNEGYKSLFKKDIIETILPEFLDDKDEIALVNSFNGFTTAFTGFFDNRENMFS
                     EEAKSTSIAFRCINENLTRYISNMDIFEKVDAIFDKHEVQEIKEKILNSDYDVEDFFEG
                     EFFNFVLTQEGIDVYNAIIGGFVTESGEKIKGLNEYINLYNQKTKQKLPKFKPLYKQVL
                     SDRESLSFYGEGYTSDEEVLEVFRNTLNKNSEIFSSIKKLEKLFKNFDEYSSAGIFVKN
                     GPAISTISKDIFGEWNVIRDKWNAEYDDIHLKKKAVVTEKYEDDRRKSFKKIGSFSLEQ
                     LQEYADADLSVVEKLKEIIIQKVDEIYKVYGSSEKLFDADFVLEKSLKKNDAVVAIMKD
                     LLDSVKSFENYIKAFFGEGKETNRDESFYGDFVLAYDILLKVDHIYDAIRNYVTQKPYS
                     KDKFKLYFQNPQFMGGWDKDKETDYRATILRYGSKYYLAIMDKKYAKCLQKIDKDDVNG
                     NYEKINYKLLPGPNKMLPKVFFSKKWMAYYNPSEDIQKIYKNGTFKKGDMFNLNDCHKL
                     IDFFKDSISRYPKWSNAYDFNFSETEKYKDIAGFYREVEEQGYKVSFESASKKEVDKLV
                     EEGKLYMFQIYNKDFSDKSHGTPNLHTMYFKLLFDENNHGQIRLSGGAELFMRRASLKK
                     EELVVHPANSPIANKNPDNPKKTTTLSYDVYKDKRFSEDQYELHIPIAINKCPKNIFKI
                     NTEVRVLLKHDDNPYVIGIARGERNLLYIVVVDGKGNIVEQYSLNEIINNFNGIRIKTD
                     YHSLLDKKEKERFEARQNWTSIENIKELKAGYISQVVHKICELVEKYDAVIALADLNSG
                     FKNSRVKVEKQVYQKFEKMLIDKLNYMVDKKSNPCATGGALKGYQITNKFESFKSMSTQ
                     NGFIFYIPAWLTSKIDPSTGFVNLLKTKYTSIADSKKFISSFDRIMYVPEEDLFEFALD
                     YKNFSRTDADYIKKWKLYSYGNRIRIFRNPKKNNVFDWEEVCLTSAYKELFNKYGINYQ
                     QGDIRALLCEQSDKAFYSSFMALMSLMLQMRNSITGRTDVAFLISPVKNSDGIFYDSRN
                     YEAQENAILPKNADANGAYNIARKVLWAIGQFKKAEDEKLDKVKIAISNKEWLEYAQTS
                     VKH"
     CDS             4768..4788
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4801..5049
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             5062..5082
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5091..5108
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5131..5148)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5137..5361
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5432..5448)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5432..5448)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5445..5467)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5456..5472
                     /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(5480..5486,5487..5504,5505..5510))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(5663..5680)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5834..6422)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5914..5933)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6593..7384,7385..7453))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLFRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7216..7235
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7454..7558)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7637..7656)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        join(7828..8076,1..131)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
       61 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      121 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      181 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      241 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      301 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
      361 agagatccgc ggccgctaat acgactcact atagggagag ccgccaccat gcccaagaag
      421 aagaggaaag tcatggaggc atctccagca agcggaccaa ggcacctgat ggacccccac
      481 atcttcacct ctaactttaa caatggcatc ggcaggcaca agacatacct gtgctatgag
      541 gtggagcgcc tggacaatgg caccagcgtg aagatggatc agcacagagg cttcctgcac
      601 aaccaggcca agaatctgct gtgcggcttc tacggccggc acgcagagct gagatttctg
      661 gacctggtgc ctagcctgca gctggatcca gcccagatct atagggtgac ctggttcatc
      721 agctactccc catgcttttc cgcgggatgt gcaggagagg tgcgcgcctt cctgcaggag
      781 aacacacacg tgcggctgag aatctttgcc gcccggatct tcgactatga tcctctgtac
      841 aaggaggccc tgcagatgct gagagacgca ggagcccagg tgtccatcat gacctatgat
      901 gagttcaagc actgctggga cacatttgtg gatcaccagg gctgtccctt tcagccttgg
      961 gacggactgg atgagcactc ccaggccctg tctggcaggc tgagggccat cctgcagaac
     1021 cagggcaata gcggcagcga gactcccggg acctcagagt ccgccacacc cgaaagtatg
     1081 agcaagctgg agaagtttac aaactgctac tccctgtcta agaccctgag gttcaaggcc
     1141 atccctgtgg gcaagaccca ggagaacatc gacaataagc ggctgctggt ggaggacgag
     1201 aagagagccg aggattataa gggcgtgaag aagctgctgg atcgctacta tctgtccttt
     1261 atcaacgacg tgctgcactc tatcaagctg aagaatctga acaattacat cagcctgttc
     1321 cggaagaaga caagaaccga gaaggagaat aaggagctgg agaacctgga gatcaatctg
     1381 cggaaggaga tcgccaaggc cttcaagggc aacgagggct acaagtccct gtttaagaag
     1441 gatatcatcg agacaatcct gccagagttc ctggacgata aggacgagat cgccctggtg
     1501 aacagcttca atggctttac cacagccttc accggcttct ttgataacag agagaatatg
     1561 ttttccgagg aggccaagag cacatccatc gccttcaggt gtatcaacga gaatctgacc
     1621 cgctacatct ctaatatgga catcttcgag aaggtggacg ccatctttga taagcacgag
     1681 gtgcaggaga tcaaggagaa gatcctgaac agcgactatg atgtggagga tttctttgag
     1741 ggcgagttct ttaactttgt gctgacacag gagggcatcg acgtgtacaa tgccatcatc
     1801 ggcggcttcg tgaccgagag cggcgagaag atcaagggcc tgaacgagta catcaacctg
     1861 tataatcaga agaccaagca gaagctgcct aagtttaagc cactgtataa gcaggtgctg
     1921 agcgatcggg agtctctgag cttctacggc gagggctata catccgatga ggaggtgctg
     1981 gaggtgttta gaaacaccct gaacaagaat tccgagatct tcagctccat caagaagctg
     2041 gagaagctgt tcaagaattt tgacgagtac tctagcgccg gcatctttgt gaagaacggc
     2101 cccgccatca gcacaatctc caaggatatc ttcggcgagt ggaacgtgat ccgggacaag
     2161 tggaatgccg agtatgacga tatccacctg aagaagaagg ccgtggtgac cgagaagtac
     2221 gaggacgatc ggagaaagtc cttcaagaag atcggctcct tttctctgga gcagctgcag
     2281 gagtacgccg acgccgatct gtctgtggtg gagaagctga aggagatcat catccagaag
     2341 gtggatgaga tctacaaggt gtatggctcc tctgagaagc tgttcgacgc cgattttgtg
     2401 ctggagaaga gcctgaagaa gaacgacgcc gtggtggcca tcatgaagga cctgctggat
     2461 tctgtgaaga gcttcgagaa ttacatcaag gccttctttg gcgagggcaa ggagacaaac
     2521 agggacgagt ccttctatgg cgattttgtg ctggcctacg acatcctgct gaaggtggac
     2581 cacatctacg atgccatccg caattatgtg acccagaagc cctactctaa ggataagttc
     2641 aagctgtatt ttcagaaccc tcagttcatg ggcggctggg acaaggataa ggagacagac
     2701 tatcgggcca ccatcctgag atacggctcc aagtactatc tggccatcat ggataagaag
     2761 tacgccaagt gcctgcagaa gatcgacaag gacgatgtga acggcaatta cgagaagatc
     2821 aactataagc tgctgcccgg ccctaataag atgctgccaa aggtgttctt ttccaagaag
     2881 tggatggcct actataaccc ctctgaggac atccagaaga tctacaagaa tggcacattc
     2941 aagaagggcg atatgtttaa cctgaatgac tgtcacaagc tgatcgactt ctttaaggat
     3001 agcatctccc ggtatccaaa gtggagcaat gcctacgatt tcaacttttc cgagaccgag
     3061 aagtataagg acatcgccgg cttttacaga gaggtggagg agcagggcta taaggtgagc
     3121 ttcgagtctg ccagcaagaa ggaggtggat aagctggtgg aggagggcaa gctgtacatg
     3181 ttccagatct ataacaagga cttttccgat aagtctcacg gcacacccaa tctgcacacc
     3241 atgtacttca agctgctgtt tgacgagaac aatcacggcc agatcagact gtctggagga
     3301 gcagagctgt tcatgaggag ggcaagcctg aagaaggagg agctggtggt gcacccagca
     3361 aactccccta tcgccaacaa gaatccagat aatcccaaga agaccacaac cctgagctac
     3421 gacgtgtata aggataagag gttttccgag gaccagtacg agctgcacat cccaatcgcc
     3481 atcaataagt gccccaagaa catcttcaag atcaatacag aggtgcgcgt gctgctgaag
     3541 cacgacgata acccctatgt gatcggcatc gccaggggcg agcgcaatct gctgtacatc
     3601 gtggtggtgg acggcaaggg caacatcgtg gagcagtatt ccctgaacga gatcatcaac
     3661 aacttcaacg gcatcaggat caagacagat taccactctc tgctggacaa gaaggagaag
     3721 gagaggttcg aggcccgcca gaactggacc tccatcgaga atatcaagga gctaaaggcc
     3781 ggctatatct ctcaggtggt gcacaagatc tgcgagctgg tggagaagta cgatgccgtg
     3841 atcgccctgg ccgacctgaa ctctggcttt aagaatagcc gcgtgaaggt ggagaagcag
     3901 gtgtatcaga agttcgagaa gatgctgatc gataagctga actacatggt ggacaagaag
     3961 tctaatcctt gtgcaacagg aggcgccctg aagggatatc agatcaccaa taagttcgag
     4021 agctttaagt ccatgtctac ccagaacggc ttcatctttt acatccctgc ctggctgaca
     4081 tccaagatcg atccatctac cggctttgtg aatctgctga agacaaagta taccagcatc
     4141 gccgattcca agaagttcat cagctccttt gacaggatca tgtacgtgcc cgaggaggat
     4201 ctgttcgagt ttgccctgga ctataagaac ttctctcgca cagacgccga ttacatcaag
     4261 aagtggaagc tgtacagcta tggcaaccgg atcagaatct tccggaatcc taagaagaac
     4321 aacgtgttcg actgggagga ggtgtgcctg accagcgcct ataaggagct gttcaacaag
     4381 tacggcatca attatcagca gggcgatatc agagccctgc tgtgcgagca gtccgacaag
     4441 gccttctact ctagctttat ggccctgatg tctctgatgc tgcagatgag gaatagcatc
     4501 acaggccgca ccgatgtggc ctttctgatc agccctgtga agaactccga cggcatcttc
     4561 tacgatagcc ggaactatga ggcccaggag aatgccatcc tgccaaagaa cgccgacgcc
     4621 aatggcgcct ataacatcgc cagaaaggtg ctgtgggcca tcggccagtt caagaaggcc
     4681 gaggacgaga agctggataa ggtgaagatc gccatctcca acaaggagtg gctggagtac
     4741 gcccagacct ctgtgaagca cggcagcccc aagaagaagc ggaaggtgtc tggtggttct
     4801 accaacctgt ctgacatcat cgagaaggag acaggcaagc agctggtcat ccaggagagc
     4861 atcctgatgc tgccagagga ggtggaggaa gtgatcggca acaagcccga gtccgatatc
     4921 ctggtgcaca ccgcctacga cgagagcaca gatgagaatg tgatgctgct gacctccgac
     4981 gcccccgagt ataagccttg ggccctggtc atccaggatt ctaacggcga gaataagatc
     5041 aagatgctgt ctggtggttc tcccaagaag aagaggaaag tctaaccggt catcatcacc
     5101 atcaccattg agtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat
     5161 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     5221 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     5281 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg
     5341 gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc tcgataccgt
     5401 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     5461 atccgctcac aattccacac aacatacgag ccgaaagcat aaagtgtaaa gcctagggtg
     5521 cctaatgagt aagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     5581 aaaacctgtc gtgccagctg cattaataaa tcggccaacg cgcggggaaa ggcggtttgc
     5641 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     5701 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     5761 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     5821 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     5881 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     5941 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     6001 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     6061 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     6121 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     6181 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     6241 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     6301 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     6361 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     6421 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     6481 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     6541 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     6601 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     6661 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     6721 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     6781 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     6841 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     6901 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     6961 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     7021 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     7081 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     7141 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     7201 cggcgtcaat acggaataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     7261 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     7321 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     7381 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     7441 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     7501 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     7561 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatcgatct cccgatcccc
     7621 tagggtcgac tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc
     7681 ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc
     7741 aaggcttgac cgacaattgc atgaagaatc tgcttagggt taggcgtttt gcgctgcttc
     7801 gcgatgtacg ggccagatat acgcgttgac attgattatt gactagttat taatagtaat
     7861 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
     7921 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
     7981 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
     8041 ggtaaactgc ccacttggca gtacatcaag tgtatc
//
