LOCUS       sequence-207550-        8142 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses dLbCpf1-BE0 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8142)
  AUTHORS   Li X, Wang Y, Liu Y, Yang B, Wang X, Wei J, Lu Z, Zhang Y, Wu J, 
            Huang X, Yang L, Chen J
  TITLE     Base editing with a Cpf1-cytidine deaminase fusion.
  JOURNAL   Nat Biotechnol. 2018 Mar 19. pii: nbt.4102. doi: 10.1038/nbt.4102.
  PUBMED    29553573
REFERENCE   2  (bases 1 to 8142)
  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 
            Biotechnol. 2018 Mar 19. pii: nbt.4102. doi: 10.1038/nbt.4102."
COMMENT     Sequence Label: pCMV-dCpf1-BE0
FEATURES             Location/Qualifiers
     source          1..8142
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        44..423
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        424..627
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     578..598
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     669..688
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        669..687
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             701..1387
                     /codon_start=1
                     /product="cytidine deaminase (C to U editing enzyme) from 
                     rat"
                     /label=APOBEC-1
                     /note="can use ssDNA as a substrate (Komor et al., 2016)"
                     /translation="MSSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWG
                     GRHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFL
                     SRYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSN
                     EAHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHI
                     LWATGLK"
     CDS             5126..5146
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5159..5407
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             5420..5440
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5449..5466
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5489..5506)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5495..5719
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5790..5806)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5790..5806)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5803..5825)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5814..5830
                     /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(5838..5844,5845..5862,5863..5868))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5883..5904
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6021..6038)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6192..6780)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6272..6291)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6951..7742,7743..7811))
                     /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     7574..7593
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7812..7916)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7995..8014)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc gttgacattg attattgact
       61 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
      121 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
      181 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
      241 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
      301 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
      361 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
      421 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga
      481 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
      541 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta
      601 cggtgggagg tctatataag cagagctggt ttagtgaacc gtcagatccg ctagagatcc
      661 gcggccgcta atacgactca ctatagggag agccgccacc atgagctcag agactggccc
      721 agtggctgtg gaccccacat tgagacggcg gatcgagccc catgagtttg aggtattctt
      781 cgatccgaga gagctccgca aggagacctg cctgctttac gaaattaatt gggggggccg
      841 gcactccatt tggcgacata catcacagaa cactaacaag cacgtcgaag tcaacttcat
      901 cgagaagttc acgacagaaa gatatttctg tccgaacaca aggtgcagca ttacctggtt
      961 tctcagctgg agcccatgcg gcgaatgtag tagggccatc actgaattcc tgtcaaggta
     1021 tccccacgtc actctgttta tttacatcgc aaggctgtac caccacgctg acccccgcaa
     1081 tcgacaaggc ctgcgggatt tgatctcttc aggtgtgact atccaaatta tgactgagca
     1141 ggagtcagga tactgctgga gaaactttgt gaattatagc ccgagtaatg aagcccactg
     1201 gcctaggtat ccccatctgt gggtacgact gtacgttctt gaactgtact gcatcatact
     1261 gggcctgcct ccttgtctca acattctgag aaggaagcag ccacagctga cattctttac
     1321 catcgctctt cagtcttgtc attaccagcg actgccccca cacattctct gggccaccgg
     1381 gttgaaaagc ggcagcgaga ctcccgggac ctcagagtcc gccacacccg aaagtatgtc
     1441 aaagctggag aaattcacca actgttatag cctgtctaag accctgcgct tcaaggcaat
     1501 cccagtgggc aagacacaag agaacattga caacaaacgg ctcctggtgg aggatgagaa
     1561 gagggctgaa gattacaagg gcgttaagaa gctgctggat aggtactatc tgtcattcat
     1621 caacgatgtc ctccacagta tcaagctgaa gaatctgaac aattacattt ctctgttccg
     1681 gaagaagaca cggaccgaga aggagaacaa agagctggag aatctggaga tcaacctgag
     1741 gaaagaaata gctaaggctt tcaaagggaa cgagggttac aagtccctgt tcaagaaaga
     1801 cattatcgag actattctgc ctgagttcct ggacgataaa gatgagatcg ccctcgtcaa
     1861 ttccttcaat gggtttacca cagcctttac cggcttcttc gacaatagag agaatatgtt
     1921 ctctgaagag gccaaatcca ctagcatcgc ctttcgctgc ataaacgaga acctgactag
     1981 gtacatcagc aatatggaca tctttgagaa agtcgatgcc atattcgaca aacatgaggt
     2041 gcaggagatt aaggagaaga tcctgaactc agattacgat gtcgaagatt tcttcgaggg
     2101 agagttcttc aacttcgtgc tcacacaaga gggcattgat gtgtacaatg caatcattgg
     2161 agggttcgtg acagagagtg gcgagaagat aaagggcctg aacgagtata tcaacctcta
     2221 caaccagaaa accaagcaga aactgcctaa gttcaagcca ctgtacaaac aagtgctctc
     2281 agatagggaa agcctgagct tctacggtga agggtataca tcagatgaag aagtgctcga
     2341 agtgttccgc aacaccctca ataagaacag tgaaatcttc tcttcaatca agaagctgga
     2401 gaaactgttc aagaatttcg atgagtactc ctctgccgga atctttgtga agaatggccc
     2461 tgcaatatcc actattagca aagacatctt tggcgagtgg aacgttatca gggataagtg
     2521 gaatgccgag tacgatgata ttcatctcaa gaagaaagcc gtggttacag agaaatacga
     2581 ggatgataga cgcaagagct ttaagaagat tggtagcttc tctctcgaac agctgcagga
     2641 gtacgccgac gctgacctgt cagtcgtgga gaaactcaag gagatcataa tccagaaggt
     2701 ggatgaaatc tacaaagtgt atggaagctc tgagaaactc ttcgatgcag actttgttct
     2761 ggagaagagt ctgaagaaga acgacgcagt ggttgctatc atgaaggacc tgctggattc
     2821 tgttaagtct ttcgagaatt acattaaggc attctttggt gaagggaagg agacaaatag
     2881 ggacgagagc ttctatggcg actttgttct ggcctacgac atcctcctca aggttgacca
     2941 catctatgac gctatacgga attacgttac ccagaagccc tatagcaaag acaagttcaa
     3001 gctgtatttc cagaatccac agtttatggg tgggtgggat aaagacaaag aaacagatta
     3061 cagggccact atcctgcggt acggcagcaa atactatctg gctatcatgg ataagaagta
     3121 cgccaaatgc ctccagaaga tcgacaagga cgacgtgaac ggtaactacg agaagatcaa
     3181 ttacaagctc ctgccaggac ctaacaagat gctgcccaag gtgttcttct ccaagaaatg
     3241 gatggcctac tataacccaa gcgaggacat tcagaagata tacaagaatg ggacattcaa
     3301 gaagggcgat atgttcaacc tcaacgactg ccacaagctg attgatttct tcaaggatag
     3361 catttctcgc tatcccaagt ggtctaatgc atacgatttc aacttcagcg agactgagaa
     3421 gtacaaagac atcgctggct tctaccggga ggtggaagag caaggctata aggtgtcatt
     3481 cgaatccgct tctaagaagg aagtggataa gctcgtggaa gagggtaagc tgtacatgtt
     3541 ccagatatac aacaaagact tcagcgataa gagccacggc actccaaacc tccatactat
     3601 gtatttcaag ctgctgtttg acgagaacaa ccacggacag attaggctgt caggaggcgc
     3661 agaactcttc atgcgcagag cttcactgaa gaaggaggaa ctcgttgtcc acccagccaa
     3721 tagccctata gccaataaga atccagacaa tcctaagaaa accactactc tgtcttacga
     3781 tgtgtataag gataagagat tctctgaaga tcagtacgaa ctgcacatac ccattgccat
     3841 taacaagtgc cctaagaaca tcttcaagat taacacagag gttagagtgc tcctgaaaca
     3901 cgacgataac ccttatgtta taggcattgc tcgcggagag agaaacctgc tgtacatcgt
     3961 cgtggtggac ggcaaaggca acatcgtgga acagtacagt ctcaatgaaa tcattaacaa
     4021 tttcaacgga atccgcatta agaccgacta ccattctctc ctcgacaaga aggagaaaga
     4081 aaggttcgaa gcaagacaga attggacaag tatagagaat atcaaagaac tgaaggctgg
     4141 gtacatctct caggttgtgc acaagatatg tgagctggtg gagaagtacg acgctgttat
     4201 cgccctcgcg gacctgaata gcggcttcaa gaactccagg gtgaaggtgg agaagcaggt
     4261 gtatcagaag ttcgagaaga tgctgatcga caagctcaac tatatggtgg acaagaaatc
     4321 caatccttgc gctactggtg gagccctgaa gggctatcaa atcaccaata agttcgaatc
     4381 tttcaagtct atgagcaccc agaatggctt catcttctac atacccgcat ggctgacatc
     4441 caagattgat ccctctaccg gatttgttaa tctgctcaag actaagtaca cctctattgc
     4501 tgactcaaag aagttcatat catcatttga ccgcatcatg tacgtgccag aagaggacct
     4561 gttcgagttt gccctggatt acaagaattt ctctcggact gacgccgact acatcaagaa
     4621 gtggaagctc tactcttatg gtaatcggat tcgcatattc cgcaatccca agaagaataa
     4681 cgtgttcgat tgggaggaag tttgcctcac cagcgcttac aaggagctgt tcaataagta
     4741 tgggattaac taccagcagg gcgacataag agccctgctg tgcgaacaat ctgataaggc
     4801 attctattcc tctttcatgg cactgatgtc actgatgctg caaatgcgca attccatcac
     4861 cggaagaaca gacgtggcct ttctgatctc tcctgtcaag aactcagatg gcatcttcta
     4921 cgattcccgc aactatgaag cacaggagaa tgctatcctg cctaagaatg ccgatgcaaa
     4981 tggagcctat aacatcgcca gaaaggtcct ctgggccata ggacaattca agaaagctga
     5041 agatgagaag ctggacaagg tgaagatcgc catttcaaac aaagagtggc tcgaatatgc
     5101 tcagacctca gtgaagcatg gatcacccaa gaagaaacgg aaagtgtctg gtggttctac
     5161 taatctgtca gatattattg aaaaggagac cggtaagcaa ctggttatcc aggaatccat
     5221 cctcatgctc ccagaggagg tggaagaagt cattgggaac aagccggaaa gcgatatact
     5281 cgtgcacacc gcctacgacg agagcaccga cgagaatgtc atgcttctga ctagcgacgc
     5341 ccctgaatac aagccttggg ctctggtcat acaggatagc aacggtgaga acaagattaa
     5401 gatgctctct ggtggttctc ccaagaagaa gaggaaagtc taaccggtca tcatcaccat
     5461 caccattgag tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     5521 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     5581 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     5641 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     5701 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc gataccgtcg
     5761 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     5821 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctagggtgcc
     5881 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     5941 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     6001 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     6061 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     6121 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     6181 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     6241 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     6301 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     6361 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     6421 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     6481 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     6541 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     6601 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     6661 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     6721 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     6781 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     6841 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     6901 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     6961 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     7021 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     7081 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     7141 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     7201 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     7261 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     7321 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     7381 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     7441 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     7501 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     7561 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     7621 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     7681 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     7741 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     7801 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     7861 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     7921 tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atcgatctcc cgatccccta
     7981 gggtcgactc tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct
     8041 gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa
     8101 ggcttgaccg acaattgcat gaagaatctg cttagggtta gg
//
