LOCUS       sequence-203055-        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"
     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             409..1095
                     /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             4834..4854
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4867..5115
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             5128..5148
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5157..5174
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5197..5214)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5203..5427
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5498..5514)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5498..5514)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5511..5533)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5522..5538
                     /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(5546..5552,5553..5570,5571..5576))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5591..5612
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5729..5746)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5900..6488)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5980..5999)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6659..7450,7451..7519))
                     /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     7282..7301
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7520..7624)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7703..7722)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        join(7894..8142,1..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 gagctcagag
      421 actggcccag tggctgtgga ccccacattg agacggcgga tcgagcccca tgagtttgag
      481 gtattcttcg atccgagaga gctccgcaag gagacctgcc tgctttacga aattaattgg
      541 gggggccggc actccatttg gcgacataca tcacagaaca ctaacaagca cgtcgaagtc
      601 aacttcatcg agaagttcac gacagaaaga tatttctgtc cgaacacaag gtgcagcatt
      661 acctggtttc tcagctggag cccatgcggc gaatgtagta gggccatcac tgaattcctg
      721 tcaaggtatc cccacgtcac tctgtttatt tacatcgcaa ggctgtacca ccacgctgac
      781 ccccgcaatc gacaaggcct gcgggatttg atctcttcag gtgtgactat ccaaattatg
      841 actgagcagg agtcaggata ctgctggaga aactttgtga attatagccc gagtaatgaa
      901 gcccactggc ctaggtatcc ccatctgtgg gtacgactgt acgttcttga actgtactgc
      961 atcatactgg gcctgcctcc ttgtctcaac attctgagaa ggaagcagcc acagctgaca
     1021 ttctttacca tcgctcttca gtcttgtcat taccagcgac tgcccccaca cattctctgg
     1081 gccaccgggt tgaaaagcgg cagcgagact cccgggacct cagagtccgc cacacccgaa
     1141 agtatgtcaa agctggagaa attcaccaac tgttatagcc tgtctaagac cctgcgcttc
     1201 aaggcaatcc cagtgggcaa gacacaagag aacattgaca acaaacggct cctggtggag
     1261 gatgagaaga gggctgaaga ttacaagggc gttaagaagc tgctggatag gtactatctg
     1321 tcattcatca acgatgtcct ccacagtatc aagctgaaga atctgaacaa ttacatttct
     1381 ctgttccgga agaagacacg gaccgagaag gagaacaaag agctggagaa tctggagatc
     1441 aacctgagga aagaaatagc taaggctttc aaagggaacg agggttacaa gtccctgttc
     1501 aagaaagaca ttatcgagac tattctgcct gagttcctgg acgataaaga tgagatcgcc
     1561 ctcgtcaatt ccttcaatgg gtttaccaca gcctttaccg gcttcttcga caatagagag
     1621 aatatgttct ctgaagaggc caaatccact agcatcgcct ttcgctgcat aaacgagaac
     1681 ctgactaggt acatcagcaa tatggacatc tttgagaaag tcgatgccat attcgacaaa
     1741 catgaggtgc aggagattaa ggagaagatc ctgaactcag attacgatgt cgaagatttc
     1801 ttcgagggag agttcttcaa cttcgtgctc acacaagagg gcattgatgt gtacaatgca
     1861 atcattggag ggttcgtgac agagagtggc gagaagataa agggcctgaa cgagtatatc
     1921 aacctctaca accagaaaac caagcagaaa ctgcctaagt tcaagccact gtacaaacaa
     1981 gtgctctcag atagggaaag cctgagcttc tacggtgaag ggtatacatc agatgaagaa
     2041 gtgctcgaag tgttccgcaa caccctcaat aagaacagtg aaatcttctc ttcaatcaag
     2101 aagctggaga aactgttcaa gaatttcgat gagtactcct ctgccggaat ctttgtgaag
     2161 aatggccctg caatatccac tattagcaaa gacatctttg gcgagtggaa cgttatcagg
     2221 gataagtgga atgccgagta cgatgatatt catctcaaga agaaagccgt ggttacagag
     2281 aaatacgagg atgatagacg caagagcttt aagaagattg gtagcttctc tctcgaacag
     2341 ctgcaggagt acgccgacgc tgacctgtca gtcgtggaga aactcaagga gatcataatc
     2401 cagaaggtgg atgaaatcta caaagtgtat ggaagctctg agaaactctt cgatgcagac
     2461 tttgttctgg agaagagtct gaagaagaac gacgcagtgg ttgctatcat gaaggacctg
     2521 ctggattctg ttaagtcttt cgagaattac attaaggcat tctttggtga agggaaggag
     2581 acaaataggg acgagagctt ctatggcgac tttgttctgg cctacgacat cctcctcaag
     2641 gttgaccaca tctatgacgc tatacggaat tacgttaccc agaagcccta tagcaaagac
     2701 aagttcaagc tgtatttcca gaatccacag tttatgggtg ggtgggataa agacaaagaa
     2761 acagattaca gggccactat cctgcggtac ggcagcaaat actatctggc tatcatggat
     2821 aagaagtacg ccaaatgcct ccagaagatc gacaaggacg acgtgaacgg taactacgag
     2881 aagatcaatt acaagctcct gccaggacct aacaagatgc tgcccaaggt gttcttctcc
     2941 aagaaatgga tggcctacta taacccaagc gaggacattc agaagatata caagaatggg
     3001 acattcaaga agggcgatat gttcaacctc aacgactgcc acaagctgat tgatttcttc
     3061 aaggatagca tttctcgcta tcccaagtgg tctaatgcat acgatttcaa cttcagcgag
     3121 actgagaagt acaaagacat cgctggcttc taccgggagg tggaagagca aggctataag
     3181 gtgtcattcg aatccgcttc taagaaggaa gtggataagc tcgtggaaga gggtaagctg
     3241 tacatgttcc agatatacaa caaagacttc agcgataaga gccacggcac tccaaacctc
     3301 catactatgt atttcaagct gctgtttgac gagaacaacc acggacagat taggctgtca
     3361 ggaggcgcag aactcttcat gcgcagagct tcactgaaga aggaggaact cgttgtccac
     3421 ccagccaata gccctatagc caataagaat ccagacaatc ctaagaaaac cactactctg
     3481 tcttacgatg tgtataagga taagagattc tctgaagatc agtacgaact gcacataccc
     3541 attgccatta acaagtgccc taagaacatc ttcaagatta acacagaggt tagagtgctc
     3601 ctgaaacacg acgataaccc ttatgttata ggcattgctc gcggagagag aaacctgctg
     3661 tacatcgtcg tggtggacgg caaaggcaac atcgtggaac agtacagtct caatgaaatc
     3721 attaacaatt tcaacggaat ccgcattaag accgactacc attctctcct cgacaagaag
     3781 gagaaagaaa ggttcgaagc aagacagaat tggacaagta tagagaatat caaagaactg
     3841 aaggctgggt acatctctca ggttgtgcac aagatatgtg agctggtgga gaagtacgac
     3901 gctgttatcg ccctcgcgga cctgaatagc ggcttcaaga actccagggt gaaggtggag
     3961 aagcaggtgt atcagaagtt cgagaagatg ctgatcgaca agctcaacta tatggtggac
     4021 aagaaatcca atccttgcgc tactggtgga gccctgaagg gctatcaaat caccaataag
     4081 ttcgaatctt tcaagtctat gagcacccag aatggcttca tcttctacat acccgcatgg
     4141 ctgacatcca agattgatcc ctctaccgga tttgttaatc tgctcaagac taagtacacc
     4201 tctattgctg actcaaagaa gttcatatca tcatttgacc gcatcatgta cgtgccagaa
     4261 gaggacctgt tcgagtttgc cctggattac aagaatttct ctcggactga cgccgactac
     4321 atcaagaagt ggaagctcta ctcttatggt aatcggattc gcatattccg caatcccaag
     4381 aagaataacg tgttcgattg ggaggaagtt tgcctcacca gcgcttacaa ggagctgttc
     4441 aataagtatg ggattaacta ccagcagggc gacataagag ccctgctgtg cgaacaatct
     4501 gataaggcat tctattcctc tttcatggca ctgatgtcac tgatgctgca aatgcgcaat
     4561 tccatcaccg gaagaacaga cgtggccttt ctgatctctc ctgtcaagaa ctcagatggc
     4621 atcttctacg attcccgcaa ctatgaagca caggagaatg ctatcctgcc taagaatgcc
     4681 gatgcaaatg gagcctataa catcgccaga aaggtcctct gggccatagg acaattcaag
     4741 aaagctgaag atgagaagct ggacaaggtg aagatcgcca tttcaaacaa agagtggctc
     4801 gaatatgctc agacctcagt gaagcatgga tcacccaaga agaaacggaa agtgtctggt
     4861 ggttctacta atctgtcaga tattattgaa aaggagaccg gtaagcaact ggttatccag
     4921 gaatccatcc tcatgctccc agaggaggtg gaagaagtca ttgggaacaa gccggaaagc
     4981 gatatactcg tgcacaccgc ctacgacgag agcaccgacg agaatgtcat gcttctgact
     5041 agcgacgccc ctgaatacaa gccttgggct ctggtcatac aggatagcaa cggtgagaac
     5101 aagattaaga tgctctctgg tggttctccc aagaagaaga ggaaagtcta accggtcatc
     5161 atcaccatca ccattgagtt taaacccgct gatcagcctc gactgtgcct tctagttgcc
     5221 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     5281 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     5341 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     5401 atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcga
     5461 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     5521 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     5581 agggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     5641 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     5701 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     5761 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     5821 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     5881 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     5941 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     6001 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     6061 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     6121 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     6181 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     6241 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     6301 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     6361 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     6421 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     6481 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     6541 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     6601 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     6661 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     6721 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     6781 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     6841 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     6901 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     6961 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     7021 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     7081 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     7141 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     7201 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     7261 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     7321 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     7381 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     7441 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     7501 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     7561 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     7621 cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat cgatctcccg
     7681 atcccctagg gtcgactctc agtacaatct gctctgatgc cgcatagtta agccagtatc
     7741 tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa
     7801 caaggcaagg cttgaccgac aattgcatga agaatctgct tagggttagg cgttttgcgc
     7861 tgcttcgcga tgtacgggcc agatatacgc gttgacattg attattgact agttattaat
     7921 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
     7981 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
     8041 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
     8101 atttacggta aactgcccac ttggcagtac atcaagtgta tc
//
