LOCUS       sequence-270339-        6207 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the N-terminal split-intein fragment of S. aureus BE3.9max
            from the CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6207)
  AUTHORS   Levy JM, Yeh WH, Pendse N, Davis JR, Hennessey E, Butcher R, Koblan 
            LW, Comander J, Liu Q, Liu DR
  TITLE     Cytosine and adenine base editing of the brain, liver, retina, heart
            and skeletal muscle of mice via adeno-associated viruses.
  JOURNAL   Nat Biomed Eng. 2020 Jan;4(1):97-110. doi: 
            10.1038/s41551-019-0501-5. Epub 2020 Jan 14.
  PUBMED    31937940
REFERENCE   2  (bases 1 to 6207)
  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"; doi: 
            "10.1038/s41551-019-0501-5"; journalName: "Nat Biomed Eng"; date: 
            "2020-01"; volume: "4"; issue: "1"; pages: "97-110"
COMMENT     Sequence Label: CMV_Npu-saBE3.9max N-terminal
FEATURES             Location/Qualifiers
     source          1..6207
                     /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             445..465
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             466..1149
                     /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="SSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWGG
                     RHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFLS
                     RYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSNE
                     AHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHIL
                     WATGLK"
     CDS             3193..3213
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3222..3239
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3262..3279)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3268..3492
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3563..3579)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3563..3579)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3576..3598)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3587..3603
                     /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(3611..3617,3618..3635,3636..3641))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3656..3677
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3794..3811)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3965..4553)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4045..4064)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4724..5515,5516..5584))
                     /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     5347..5366
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5585..5689)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5768..5787)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        join(5959..6207,1..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 gaaacggaca
      421 gccgacggaa gcgagttcga gtcaccaaag aagaagcgga aagtctcctc agagactggg
      481 cctgtcgccg tcgatccaac cctgcgccgc cggattgaac ctcacgagtt tgaagtgttc
      541 tttgaccccc gggagctgag aaaggagaca tgcctgctgt acgagatcaa ctggggaggc
      601 aggcactcca tctggaggca cacctctcag aacacaaata agcacgtgga ggtgaacttc
      661 atcgagaagt ttaccacaga gcggtacttc tgccccaata ccagatgtag catcacatgg
      721 tttctgagct ggtccccttg cggagagtgt agcagggcca tcaccgagtt cctgtccaga
      781 tatccacacg tgacactgtt tatctacatc gccaggctgt atcaccacgc agacccaagg
      841 aataggcagg gcctgcgcga tctgatcagc tccggcgtga ccatccagat catgacagag
      901 caggagtccg gctactgctg gcggaacttc gtgaattatt ctcctagcaa cgaggcccac
      961 tggcctaggt acccacacct gtgggtgcgc ctgtacgtgc tggagctgta ttgcatcatc
     1021 ctgggcctgc ccccttgtct gaatatcctg cggagaaagc agccccagct gaccttcttt
     1081 acaatcgccc tgcagtcttg tcactatcag aggctgccac cccacatcct gtgggccaca
     1141 ggcctgaagt ctggaggatc tagcggagga tcctctggca gcgagacacc aggaacaagc
     1201 gagtcagcaa caccagagag cagtggcggc agcagcggcg gcagcgggaa gcgaaattac
     1261 attctggggc tggccattgg cattacatca gtgggctatg gcatcattga ctacgagaca
     1321 agggacgtga tcgacgccgg cgtgagactg ttcaaggagg ccaacgtgga gaacaatgag
     1381 ggccggagat ccaagagggg agcaaggcgc ctgaagcgga gaaggcgcca cagaatccag
     1441 agagtgaaga agctgctgtt cgattacaac ctgctgaccg accactccga gctgtctggc
     1501 atcaatcctt atgaggccag agtgaagggc ctgtcccaga agctgtctga ggaggagttt
     1561 agcgccgccc tgctgcacct ggcaaagagg agaggcgtgc acaacgtgaa tgaggtggag
     1621 gaggacaccg gcaacgagct gtccacaaag gagcagatca gccgcaattc caaggccctg
     1681 gaggagaagt atgtggccga gctgcagctg gagcggctga agaaggatgg cgaggtgagg
     1741 ggctccatca atcgcttcaa gacctctgac tacgtgaagg aggccaagca gctgctgaag
     1801 gtgcagaagg cctaccacca gctggatcag tcctttatcg atacatatat cgacctgctg
     1861 gagacaaggc gcacatacta tgagggacca ggagagggct ctcccttcgg ctggaaggac
     1921 atcaaggagt ggtacgagat gctgatgggc cactgcacct attttccaga ggagctgaga
     1981 agcgtgaagt acgcctataa cgccgatctg tacaacgccc tgaatgacct gaacaacctg
     2041 gtcatcacca gggatgagaa cgagaagctg gagtactatg agaagttcca gatcatcgag
     2101 aacgtgttca agcagaagaa gaagcctaca ctgaagcaga tcgccaagga gatcctggtg
     2161 aacgaggagg acatcaaggg ctaccgcgtg acctccacag gcaagccaga gttcaccaat
     2221 ctgaaggtgt atcacgatat caaggacatc acagcccgga aggagatcat cgagaacgcc
     2281 gagctgctgg atcagatcgc caagatcctg accatctatc agagctccga ggacatccag
     2341 gaggagctga ccaacctgaa tagcgagctg acacaggagg agatcgagca gatcagcaat
     2401 ctgaagggct acaccggcac acacaacctg agcctgaagg ccatcaatct gatcctggat
     2461 gagctgtggc acacaaacga caatcagatc gccatcttta accggctgaa gctggtgcca
     2521 aagaaggtgg acctgtccca gcagaaggag atcccaacca cactggtgga cgatttcatc
     2581 ctgtctcccg tggtgaagcg gagcttcatc cagagcatca aagtgatcaa cgccatcatc
     2641 aagaagtacg gcctgcccaa tgatatcatc atcgagctgg ccagggagaa gaactccaag
     2701 gacgcccaga agatgatcaa tgagatgcag aagaggaacc gccagaccaa tgagcggatc
     2761 gaggagatca tcagaaccac aggcaaggag aacgccaagt acctgatcga gaagatcaag
     2821 ctgcacgata tgcaggaggg caagtgcctg tcctacgaga cagagatcct gacagtggag
     2881 tatggcctgc tgccaatcgg caagatcgtg gagaagagga tcgagtgtac cgtgtactct
     2941 gtggataaca atggcaacat ctatacacag cccgtggcac agtggcacga taggggagag
     3001 caggaggtgt tcgagtattg cctggaggac ggcagcctga tcagggcaac caaggaccac
     3061 aagttcatga cagtggatgg ccagatgctg cccatcgacg agattttcga gcgggagctg
     3121 gacctgatga gagtggataa cctgcctaat agcggaggca gtaaaagaac agcagacggg
     3181 agtgagtttg agcccaagaa aaagagaaag gtgtaaccgg tcatcatcac catcaccatt
     3241 gagtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
     3301 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     3361 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     3421 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
     3481 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctcgataccg tcgacctcta
     3541 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     3601 caattccaca caacatacga gccggaagca taaagtgtaa agcctagggt gcctaatgag
     3661 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     3721 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     3781 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     3841 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     3901 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     3961 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     4021 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     4081 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     4141 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     4201 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     4261 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     4321 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     4381 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     4441 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     4501 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     4561 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     4621 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     4681 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     4741 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     4801 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     4861 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     4921 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     4981 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     5041 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     5101 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     5161 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     5221 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     5281 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     5341 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     5401 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     5461 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     5521 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     5581 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     5641 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     5701 gaaaagtgcc acctgacgtc gacggatcgg gagatcgatc tcccgatccc ctagggtcga
     5761 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg
     5821 tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga
     5881 ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac
     5941 gggccagata tacgcgttga cattgattat tgactagtta ttaatagtaa tcaattacgg
     6001 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
     6061 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
     6121 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
     6181 cccacttggc agtacatcaa gtgtatc
//
