LOCUS       sequence-207602-        8166 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses dLbCpf1-BE-YEE in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8166)
  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 8166)
  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-BE-YEE
FEATURES             Location/Qualifiers
     source          1..8166
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        6..385
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..589
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     540..560
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     631..650
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        631..649
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             666..686
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5112..5132
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5145..5393
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             5406..5426
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5435..5452
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5475..5492)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5481..5705
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5776..5792)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5776..5792)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5789..5811)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5800..5816
                     /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(5824..5830,5831..5848,5849..5854))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5869..5890
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6007..6024)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6178..6766)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6258..6277)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6937..7728,7729..7797))
                     /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     7560..7579
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7798..7902)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7981..8000)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gcgttgacat tgattattga ctagttatta atagtaatca attacggggt cattagttca
       61 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
      121 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
      181 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
      241 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
      301 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
      361 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
      421 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
      481 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
      541 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa
      601 ccgtcagatc cgctagagat ccgcggccgc taatacgact cactataggg agagccgcca
      661 ccatgcccaa gaagaagagg aaagtcatga gctcagagac tggcccagtg gctgtggacc
      721 ccacattgag acggcggatc gagccccatg agtttgaggt attcttcgat ccgagagagc
      781 tccgcaagga gacctgcctg ctttacgaaa ttaattgggg gggccggcac tccatttggc
      841 gacatacatc acagaacact aacaagcacg tcgaagtcaa cttcatcgag aagttcacga
      901 cagaaagata tttctgtccg aacacaaggt gcagcattac ctggtttctc agctacagcc
      961 catgcggcga atgtagtagg gccatcactg aattcctgtc aaggtatccc cacgtcactc
     1021 tgtttattta catcgcaagg ctgtaccacc acgctgaccc cgaaaatcga caaggcctgg
     1081 aggatttgat ctcttcaggt gtgactatcc aaattatgac tgagcaggag tcaggatact
     1141 gctggagaaa ctttgtgaat tatagcccga gtaatgaagc ccactggcct aggtatcccc
     1201 atctgtgggt acgactgtac gttcttgaac tgtactgcat catactgggc ctgcctcctt
     1261 gtctcaacat tctgagaagg aagcagccac agctgacatt ctttaccatc gctcttcagt
     1321 cttgtcatta ccagcgactg cccccacaca ttctctgggc caccgggttg aaaagcggca
     1381 gcgagactcc cgggacctca gagtccgcca cacccgaaag tatgtcaaag ctggagaaat
     1441 tcaccaactg ttatagcctg tctaagaccc tgcgcttcaa ggcaatccca gtgggcaaga
     1501 cacaagagaa cattgacaac aaacggctcc tggtggagga tgagaagagg gctgaagatt
     1561 acaagggcgt taagaagctg ctggataggt actatctgtc attcatcaac gatgtcctcc
     1621 acagtatcaa gctgaagaat ctgaacaatt acatttctct gttccggaag aagacacgga
     1681 ccgagaagga gaacaaagag ctggagaatc tggagatcaa cctgaggaaa gaaatagcta
     1741 aggctttcaa agggaacgag ggttacaagt ccctgttcaa gaaagacatt atcgagacta
     1801 ttctgcctga gttcctggac gataaagatg agatcgccct cgtcaattcc ttcaatgggt
     1861 ttaccacagc ctttaccggc ttcttcgaca atagagagaa tatgttctct gaagaggcca
     1921 aatccactag catcgccttt cgctgcataa acgagaacct gactaggtac atcagcaata
     1981 tggacatctt tgagaaagtc gatgccatat tcgacaaaca tgaggtgcag gagattaagg
     2041 agaagatcct gaactcagat tacgatgtcg aagatttctt cgagggagag ttcttcaact
     2101 tcgtgctcac acaagagggc attgatgtgt acaatgcaat cattggaggg ttcgtgacag
     2161 agagtggcga gaagataaag ggcctgaacg agtatatcaa cctctacaac cagaaaacca
     2221 agcagaaact gcctaagttc aagccactgt acaaacaagt gctctcagat agggaaagcc
     2281 tgagcttcta cggtgaaggg tatacatcag atgaagaagt gctcgaagtg ttccgcaaca
     2341 ccctcaataa gaacagtgaa atcttctctt caatcaagaa gctggagaaa ctgttcaaga
     2401 atttcgatga gtactcctct gccggaatct ttgtgaagaa tggccctgca atatccacta
     2461 ttagcaaaga catctttggc gagtggaacg ttatcaggga taagtggaat gccgagtacg
     2521 atgatattca tctcaagaag aaagccgtgg ttacagagaa atacgaggat gatagacgca
     2581 agagctttaa gaagattggt agcttctctc tcgaacagct gcaggagtac gccgacgctg
     2641 acctgtcagt cgtggagaaa ctcaaggaga tcataatcca gaaggtggat gaaatctaca
     2701 aagtgtatgg aagctctgag aaactcttcg atgcagactt tgttctggag aagagtctga
     2761 agaagaacga cgcagtggtt gctatcatga aggacctgct ggattctgtt aagtctttcg
     2821 agaattacat taaggcattc tttggtgaag ggaaggagac aaatagggac gagagcttct
     2881 atggcgactt tgttctggcc tacgacatcc tcctcaaggt tgaccacatc tatgacgcta
     2941 tacggaatta cgttacccag aagccctata gcaaagacaa gttcaagctg tatttccaga
     3001 atccacagtt tatgggtggg tgggataaag acaaagaaac agattacagg gccactatcc
     3061 tgcggtacgg cagcaaatac tatctggcta tcatggataa gaagtacgcc aaatgcctcc
     3121 agaagatcga caaggacgac gtgaacggta actacgagaa gatcaattac aagctcctgc
     3181 caggacctaa caagatgctg cccaaggtgt tcttctccaa gaaatggatg gcctactata
     3241 acccaagcga ggacattcag aagatataca agaatgggac attcaagaag ggcgatatgt
     3301 tcaacctcaa cgactgccac aagctgattg atttcttcaa ggatagcatt tctcgctatc
     3361 ccaagtggtc taatgcatac gatttcaact tcagcgagac tgagaagtac aaagacatcg
     3421 ctggcttcta ccgggaggtg gaagagcaag gctataaggt gtcattcgaa tccgcttcta
     3481 agaaggaagt ggataagctc gtggaagagg gtaagctgta catgttccag atatacaaca
     3541 aagacttcag cgataagagc cacggcactc caaacctcca tactatgtat ttcaagctgc
     3601 tgtttgacga gaacaaccac ggacagatta ggctgtcagg aggcgcagaa ctcttcatgc
     3661 gcagagcttc actgaagaag gaggaactcg ttgtccaccc agccaatagc cctatagcca
     3721 ataagaatcc agacaatcct aagaaaacca ctactctgtc ttacgatgtg tataaggata
     3781 agagattctc tgaagatcag tacgaactgc acatacccat tgccattaac aagtgcccta
     3841 agaacatctt caagattaac acagaggtta gagtgctcct gaaacacgac gataaccctt
     3901 atgttatagg cattgctcgc ggagagagaa acctgctgta catcgtcgtg gtggacggca
     3961 aaggcaacat cgtggaacag tacagtctca atgaaatcat taacaatttc aacggaatcc
     4021 gcattaagac cgactaccat tctctcctcg acaagaagga gaaagaaagg ttcgaagcaa
     4081 gacagaattg gacaagtata gagaatatca aagaactgaa ggctgggtac atctctcagg
     4141 ttgtgcacaa gatatgtgag ctggtggaga agtacgacgc tgttatcgcc ctcgcggacc
     4201 tgaatagcgg cttcaagaac tccagggtga aggtggagaa gcaggtgtat cagaagttcg
     4261 agaagatgct gatcgacaag ctcaactata tggtggacaa gaaatccaat ccttgcgcta
     4321 ctggtggagc cctgaagggc tatcaaatca ccaataagtt cgaatctttc aagtctatga
     4381 gcacccagaa tggcttcatc ttctacatac ccgcatggct gacatccaag attgatccct
     4441 ctaccggatt tgttaatctg ctcaagacta agtacacctc tattgctgac tcaaagaagt
     4501 tcatatcatc atttgaccgc atcatgtacg tgccagaaga ggacctgttc gagtttgccc
     4561 tggattacaa gaatttctct cggactgacg ccgactacat caagaagtgg aagctctact
     4621 cttatggtaa tcggattcgc atattccgca atcccaagaa gaataacgtg ttcgattggg
     4681 aggaagtttg cctcaccagc gcttacaagg agctgttcaa taagtatggg attaactacc
     4741 agcagggcga cataagagcc ctgctgtgcg aacaatctga taaggcattc tattcctctt
     4801 tcatggcact gatgtcactg atgctgcaaa tgcgcaattc catcaccgga agaacagacg
     4861 tggcctttct gatctctcct gtcaagaact cagatggcat cttctacgat tcccgcaact
     4921 atgaagcaca ggagaatgct atcctgccta agaatgccga tgcaaatgga gcctataaca
     4981 tcgccagaaa ggtcctctgg gccataggac aattcaagaa agctgaagat gagaagctgg
     5041 acaaggtgaa gatcgccatt tcaaacaaag agtggctcga atatgctcag acctcagtga
     5101 agcatggatc acccaagaag aaacggaaag tgtctggtgg ttctactaat ctgtcagata
     5161 ttattgaaaa ggagaccggt aagcaactgg ttatccagga atccatcctc atgctcccag
     5221 aggaggtgga agaagtcatt gggaacaagc cggaaagcga tatactcgtg cacaccgcct
     5281 acgacgagag caccgacgag aatgtcatgc ttctgactag cgacgcccct gaatacaagc
     5341 cttgggctct ggtcatacag gatagcaacg gtgagaacaa gattaagatg ctctctggtg
     5401 gttctcccaa gaagaagagg aaagtctaac cggtcatcat caccatcacc attgagttta
     5461 aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     5521 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     5581 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     5641 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     5701 tatggcttct gaggcggaaa gaaccagctg gggctcgata ccgtcgacct ctagctagag
     5761 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     5821 acacaacata cgagccggaa gcataaagtg taaagcctag ggtgcctaat gagtgagcta
     5881 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     5941 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     6001 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     6061 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     6121 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     6181 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     6241 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     6301 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     6361 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     6421 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     6481 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     6541 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     6601 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     6661 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     6721 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     6781 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     6841 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     6901 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     6961 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     7021 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     7081 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     7141 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     7201 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     7261 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     7321 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     7381 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     7441 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     7501 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     7561 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     7621 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     7681 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     7741 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     7801 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     7861 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     7921 gccacctgac gtcgacggat cgggagatcg atctcccgat cccctagggt cgactctcag
     7981 tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga
     8041 ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa
     8101 ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag
     8161 atatac
//
