LOCUS       Exported                8166 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Expresses dLbCpf1-BE-YE in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    pCMV-dCpf1-BE-YE
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 May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8166
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        16..395
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        396..599
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     550..570
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     641..660
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        641..659
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             676..696
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5122..5142
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5155..5403
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             5416..5436
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5445..5462
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5485..5502)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5491..5715
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5786..5802)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5786..5802)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5799..5821)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5810..5826
                     /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(5834..5864)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5879..5900
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6017..6034)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6188..6776)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6268..6287)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6947..7807)
                     /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     7570..7589
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7808..7912)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7991..8010)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ccagatatac gcgttgacat tgattattga ctagttatta atagtaatca attacggggt
       61 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
      121 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
      181 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
      241 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
      301 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
      361 agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca
      421 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca
      481 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg
      541 ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg
      601 gtttagtgaa ccgtcagatc cgctagagat ccgcggccgc taatacgact cactataggg
      661 agagccgcca ccatgcccaa gaagaagagg aaagtcatga gctcagagac tggcccagtg
      721 gctgtggacc ccacattgag acggcggatc gagccccatg agtttgaggt attcttcgat
      781 ccgagagagc tccgcaagga gacctgcctg ctttacgaaa ttaattgggg gggccggcac
      841 tccatttggc gacatacatc acagaacact aacaagcacg tcgaagtcaa cttcatcgag
      901 aagttcacga cagaaagata tttctgtccg aacacaaggt gcagcattac ctggtttctc
      961 agctacagcc catgcggcga atgtagtagg gccatcactg aattcctgtc aaggtatccc
     1021 cacgtcactc tgtttattta catcgcaagg ctgtaccacc acgctgaccc cgaaaatcga
     1081 caaggcctgc gggatttgat ctcttcaggt gtgactatcc aaattatgac tgagcaggag
     1141 tcaggatact gctggagaaa ctttgtgaat tatagcccga gtaatgaagc ccactggcct
     1201 aggtatcccc atctgtgggt acgactgtac gttcttgaac tgtactgcat catactgggc
     1261 ctgcctcctt gtctcaacat tctgagaagg aagcagccac agctgacatt ctttaccatc
     1321 gctcttcagt cttgtcatta ccagcgactg cccccacaca ttctctgggc caccgggttg
     1381 aaaagcggca gcgagactcc cgggacctca gagtccgcca cacccgaaag tatgtcaaag
     1441 ctggagaaat tcaccaactg ttatagcctg tctaagaccc tgcgcttcaa ggcaatccca
     1501 gtgggcaaga cacaagagaa cattgacaac aaacggctcc tggtggagga tgagaagagg
     1561 gctgaagatt acaagggcgt taagaagctg ctggataggt actatctgtc attcatcaac
     1621 gatgtcctcc acagtatcaa gctgaagaat ctgaacaatt acatttctct gttccggaag
     1681 aagacacgga ccgagaagga gaacaaagag ctggagaatc tggagatcaa cctgaggaaa
     1741 gaaatagcta aggctttcaa agggaacgag ggttacaagt ccctgttcaa gaaagacatt
     1801 atcgagacta ttctgcctga gttcctggac gataaagatg agatcgccct cgtcaattcc
     1861 ttcaatgggt ttaccacagc ctttaccggc ttcttcgaca atagagagaa tatgttctct
     1921 gaagaggcca aatccactag catcgccttt cgctgcataa acgagaacct gactaggtac
     1981 atcagcaata tggacatctt tgagaaagtc gatgccatat tcgacaaaca tgaggtgcag
     2041 gagattaagg agaagatcct gaactcagat tacgatgtcg aagatttctt cgagggagag
     2101 ttcttcaact tcgtgctcac acaagagggc attgatgtgt acaatgcaat cattggaggg
     2161 ttcgtgacag agagtggcga gaagataaag ggcctgaacg agtatatcaa cctctacaac
     2221 cagaaaacca agcagaaact gcctaagttc aagccactgt acaaacaagt gctctcagat
     2281 agggaaagcc tgagcttcta cggtgaaggg tatacatcag atgaagaagt gctcgaagtg
     2341 ttccgcaaca ccctcaataa gaacagtgaa atcttctctt caatcaagaa gctggagaaa
     2401 ctgttcaaga atttcgatga gtactcctct gccggaatct ttgtgaagaa tggccctgca
     2461 atatccacta ttagcaaaga catctttggc gagtggaacg ttatcaggga taagtggaat
     2521 gccgagtacg atgatattca tctcaagaag aaagccgtgg ttacagagaa atacgaggat
     2581 gatagacgca agagctttaa gaagattggt agcttctctc tcgaacagct gcaggagtac
     2641 gccgacgctg acctgtcagt cgtggagaaa ctcaaggaga tcataatcca gaaggtggat
     2701 gaaatctaca aagtgtatgg aagctctgag aaactcttcg atgcagactt tgttctggag
     2761 aagagtctga agaagaacga cgcagtggtt gctatcatga aggacctgct ggattctgtt
     2821 aagtctttcg agaattacat taaggcattc tttggtgaag ggaaggagac aaatagggac
     2881 gagagcttct atggcgactt tgttctggcc tacgacatcc tcctcaaggt tgaccacatc
     2941 tatgacgcta tacggaatta cgttacccag aagccctata gcaaagacaa gttcaagctg
     3001 tatttccaga atccacagtt tatgggtggg tgggataaag acaaagaaac agattacagg
     3061 gccactatcc tgcggtacgg cagcaaatac tatctggcta tcatggataa gaagtacgcc
     3121 aaatgcctcc agaagatcga caaggacgac gtgaacggta actacgagaa gatcaattac
     3181 aagctcctgc caggacctaa caagatgctg cccaaggtgt tcttctccaa gaaatggatg
     3241 gcctactata acccaagcga ggacattcag aagatataca agaatgggac attcaagaag
     3301 ggcgatatgt tcaacctcaa cgactgccac aagctgattg atttcttcaa ggatagcatt
     3361 tctcgctatc ccaagtggtc taatgcatac gatttcaact tcagcgagac tgagaagtac
     3421 aaagacatcg ctggcttcta ccgggaggtg gaagagcaag gctataaggt gtcattcgaa
     3481 tccgcttcta agaaggaagt ggataagctc gtggaagagg gtaagctgta catgttccag
     3541 atatacaaca aagacttcag cgataagagc cacggcactc caaacctcca tactatgtat
     3601 ttcaagctgc tgtttgacga gaacaaccac ggacagatta ggctgtcagg aggcgcagaa
     3661 ctcttcatgc gcagagcttc actgaagaag gaggaactcg ttgtccaccc agccaatagc
     3721 cctatagcca ataagaatcc agacaatcct aagaaaacca ctactctgtc ttacgatgtg
     3781 tataaggata agagattctc tgaagatcag tacgaactgc acatacccat tgccattaac
     3841 aagtgcccta agaacatctt caagattaac acagaggtta gagtgctcct gaaacacgac
     3901 gataaccctt atgttatagg cattgctcgc ggagagagaa acctgctgta catcgtcgtg
     3961 gtggacggca aaggcaacat cgtggaacag tacagtctca atgaaatcat taacaatttc
     4021 aacggaatcc gcattaagac cgactaccat tctctcctcg acaagaagga gaaagaaagg
     4081 ttcgaagcaa gacagaattg gacaagtata gagaatatca aagaactgaa ggctgggtac
     4141 atctctcagg ttgtgcacaa gatatgtgag ctggtggaga agtacgacgc tgttatcgcc
     4201 ctcgcggacc tgaatagcgg cttcaagaac tccagggtga aggtggagaa gcaggtgtat
     4261 cagaagttcg agaagatgct gatcgacaag ctcaactata tggtggacaa gaaatccaat
     4321 ccttgcgcta ctggtggagc cctgaagggc tatcaaatca ccaataagtt cgaatctttc
     4381 aagtctatga gcacccagaa tggcttcatc ttctacatac ccgcatggct gacatccaag
     4441 attgatccct ctaccggatt tgttaatctg ctcaagacta agtacacctc tattgctgac
     4501 tcaaagaagt tcatatcatc atttgaccgc atcatgtacg tgccagaaga ggacctgttc
     4561 gagtttgccc tggattacaa gaatttctct cggactgacg ccgactacat caagaagtgg
     4621 aagctctact cttatggtaa tcggattcgc atattccgca atcccaagaa gaataacgtg
     4681 ttcgattggg aggaagtttg cctcaccagc gcttacaagg agctgttcaa taagtatggg
     4741 attaactacc agcagggcga cataagagcc ctgctgtgcg aacaatctga taaggcattc
     4801 tattcctctt tcatggcact gatgtcactg atgctgcaaa tgcgcaattc catcaccgga
     4861 agaacagacg tggcctttct gatctctcct gtcaagaact cagatggcat cttctacgat
     4921 tcccgcaact atgaagcaca ggagaatgct atcctgccta agaatgccga tgcaaatgga
     4981 gcctataaca tcgccagaaa ggtcctctgg gccataggac aattcaagaa agctgaagat
     5041 gagaagctgg acaaggtgaa gatcgccatt tcaaacaaag agtggctcga atatgctcag
     5101 acctcagtga agcatggatc acccaagaag aaacggaaag tgtctggtgg ttctactaat
     5161 ctgtcagata ttattgaaaa ggagaccggt aagcaactgg ttatccagga atccatcctc
     5221 atgctcccag aggaggtgga agaagtcatt gggaacaagc cggaaagcga tatactcgtg
     5281 cacaccgcct acgacgagag caccgacgag aatgtcatgc ttctgactag cgacgcccct
     5341 gaatacaagc cttgggctct ggtcatacag gatagcaacg gtgagaacaa gattaagatg
     5401 ctctctggtg gttctcccaa gaagaagagg aaagtctaac cggtcatcat caccatcacc
     5461 attgagttta aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
     5521 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     5581 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     5641 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
     5701 cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctcgata ccgtcgacct
     5761 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     5821 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctag ggtgcctaat
     5881 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     5941 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     6001 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     6061 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     6121 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     6181 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     6241 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     6301 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     6361 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     6421 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     6481 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     6541 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     6601 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     6661 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     6721 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     6781 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     6841 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     6901 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     6961 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     7021 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     7081 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     7141 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     7201 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     7261 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     7321 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     7381 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     7441 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     7501 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     7561 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     7621 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     7681 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     7741 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     7801 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     7861 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     7921 cccgaaaagt gccacctgac gtcgacggat cgggagatcg atctcccgat cccctagggt
     7981 cgactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
     8041 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
     8101 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
     8161 tacggg
//
