LOCUS       sequence-203061-        8121 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses pCMV-dLbCpf1-BE0-Delta-iNLS crRNA in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8121)
  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 8121)
  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-dLbCpf1-BE0-Delta-iNLS
FEATURES             Location/Qualifiers
     source          1..8121
                     /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             4846..5094
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             5107..5127
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5136..5153
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5176..5193)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5182..5406
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5477..5493)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5477..5493)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5490..5512)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5501..5517
                     /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(5525..5531,5532..5549,5550..5555))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5570..5591
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5708..5725)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5879..6467)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5959..5978)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6638..7429,7430..7498))
                     /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     7261..7280
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7499..7603)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7682..7701)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        join(7873..8121,1..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 tcatctggtg gttctactaa tctgtcagat
     4861 attattgaaa aggagaccgg taagcaactg gttatccagg aatccatcct catgctccca
     4921 gaggaggtgg aagaagtcat tgggaacaag ccggaaagcg atatactcgt gcacaccgcc
     4981 tacgacgaga gcaccgacga gaatgtcatg cttctgacta gcgacgcccc tgaatacaag
     5041 ccttgggctc tggtcataca ggatagcaac ggtgagaaca agattaagat gctctctggt
     5101 ggttctccca agaagaagag gaaagtctaa ccggtcatca tcaccatcac cattgagttt
     5161 aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
     5221 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     5281 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     5341 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct
     5401 ctatggcttc tgaggcggaa agaaccagct ggggctcgat accgtcgacc tctagctaga
     5461 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     5521 cacacaacat acgagccgga agcataaagt gtaaagccta gggtgcctaa tgagtgagct
     5581 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     5641 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     5701 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     5761 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     5821 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     5881 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     5941 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     6001 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     6061 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     6121 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     6181 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     6241 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     6301 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     6361 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     6421 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     6481 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     6541 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     6601 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     6661 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     6721 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     6781 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     6841 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     6901 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     6961 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     7021 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     7081 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     7141 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     7201 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     7261 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     7321 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     7381 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     7441 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     7501 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     7561 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     7621 tgccacctga cgtcgacgga tcgggagatc gatctcccga tcccctaggg tcgactctca
     7681 gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg
     7741 aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca
     7801 attgcatgaa gaatctgctt agggttaggc gttttgcgct gcttcgcgat gtacgggcca
     7861 gatatacgcg ttgacattga ttattgacta gttattaata gtaatcaatt acggggtcat
     7921 tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg
     7981 gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa
     8041 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact
     8101 tggcagtaca tcaagtgtat c
//
