LOCUS       sequence-164328-        7689 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SaBE3 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7689)
  AUTHORS   Kim YB, Komor AC, Levy JM, Packer MS, Zhao KT, Liu DR
  TITLE     Increasing the genome-targeting scope and precision of base editing 
            with engineered Cas9-cytidine deaminase fusions.
  JOURNAL   Nat Biotechnol. 2017 Feb 13. doi: 10.1038/nbt.3803.
  PUBMED    28191901
REFERENCE   2  (bases 1 to 7689)
  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. 2017 Feb 13. doi: 10.1038/nbt.3803."
COMMENT     Sequence Label: pJL-SaBE3
FEATURES             Location/Qualifiers
     source          1..7689
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             13..261
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             274..294
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             303..320
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(343..360)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    349..573
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(644..660)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(644..660)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(657..679)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    668..684
                     /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(692..698,699..716,717..722))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    737..758
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(875..892)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1046..1634)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1126..1145)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1805..2596,2597..2665))
                     /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     2428..2447
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2666..2770)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2849..2868)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3040..3419
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3420..3623
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3574..3594
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     3665..3684
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3665..3683
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3697..4383
                     /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             7597..7617
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7624..7689
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
ORIGIN
        1 tctggtggtt ctactaatct gtcagatatt attgaaaagg agaccggtaa gcaactggtt
       61 atccaggaat ccatcctcat gctcccagag gaggtggaag aagtcattgg gaacaagccg
      121 gaaagcgata tactcgtgca caccgcctac gacgagagca ccgacgagaa tgtcatgctt
      181 ctgactagcg acgcccctga atacaagcct tgggctctgg tcatacagga tagcaacggt
      241 gagaacaaga ttaagatgct ctctggtggt tctcccaaga agaagaggaa agtctaaccg
      301 gtcatcatca ccatcaccat tgagtttaaa cccgctgatc agcctcgact gtgccttcta
      361 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
      421 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
      481 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
      541 gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg
      601 gctcgatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
      661 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
      721 aagcctaggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
      781 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
      841 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
      901 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
      961 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     1021 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     1081 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     1141 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     1201 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     1261 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     1321 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     1381 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     1441 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     1501 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     1561 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     1621 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     1681 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     1741 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     1801 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     1861 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     1921 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     1981 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     2041 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     2101 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     2161 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     2221 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     2281 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     2341 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     2401 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     2461 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     2521 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     2581 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     2641 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     2701 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     2761 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatcgat
     2821 ctcccgatcc cctagggtcg actctcagta caatctgctc tgatgccgca tagttaagcc
     2881 agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag
     2941 ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg gttaggcgtt
     3001 ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta ttgactagtt
     3061 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
     3121 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
     3181 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
     3241 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
     3301 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
     3361 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
     3421 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
     3481 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
     3541 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
     3601 gggaggtcta tataagcaga gctggtttag tgaaccgtca gatccgctag agatccgcgg
     3661 ccgctaatac gactcactat agggagagcc gccaccatga gctcagagac tggcccagtg
     3721 gctgtggacc ccacattgag acggcggatc gagccccatg agtttgaggt attcttcgat
     3781 ccgagagagc tccgcaagga gacctgcctg ctttacgaaa ttaattgggg gggccggcac
     3841 tccatttggc gacatacatc acagaacact aacaagcacg tcgaagtcaa cttcatcgag
     3901 aagttcacga cagaaagata tttctgtccg aacacaaggt gcagcattac ctggtttctc
     3961 agctggagcc catgcggcga atgtagtagg gccatcactg aattcctgtc aaggtatccc
     4021 cacgtcactc tgtttattta catcgcaagg ctgtaccacc acgctgaccc ccgcaatcga
     4081 caaggcctgc gggatttgat ctcttcaggt gtgactatcc aaattatgac tgagcaggag
     4141 tcaggatact gctggagaaa ctttgtgaat tatagcccga gtaatgaagc ccactggcct
     4201 aggtatcccc atctgtgggt acgactgtac gttcttgaac tgtactgcat catactgggc
     4261 ctgcctcctt gtctcaacat tctgagaagg aagcagccac agctgacatt ctttaccatc
     4321 gctcttcagt cttgtcatta ccagcgactg cccccacaca ttctctgggc caccgggttg
     4381 aaaagcggca gcgagactcc cgggacctca gagtccgcca cacccgaaag tgggaaacgg
     4441 aactacatcc tggggcttgc cattgggata accagcgttg gctacggaat tattgattat
     4501 gagacacgcg atgtgattga cgccggggtt aggctgttca aagaggccaa cgttgaaaac
     4561 aacgagggaa gacggagtaa gcgcggagca agaagactca agcgcagacg gagacatcgg
     4621 attcagaggg tgaaaaagct gctcttcgat tacaatctcc tgaccgatca tagtgagctg
     4681 agcggaatca acccctacga ggcgcgagtg aaagggcttt cccagaagct gtccgaagag
     4741 gagttctccg ccgcgttgct gcacctggcc aaacggaggg gggttcacaa tgtaaacgaa
     4801 gtggaggagg acacgggcaa tgaacttagt acgaaagaac agatcagtag gaactctaag
     4861 gctctcgaag agaaatacgt cgctgagttg cagcttgaga gactgaaaaa agacggcgaa
     4921 gtacgcggat ctattaatag gttcaagact tcagattacg taaaggaagc caagcagctc
     4981 ctgaaagtac agaaagcgta ccatcagctc gatcagagct tcatcgatac ctacatagat
     5041 ttgctggaga cacggaggac atactacgag ggcccagggg aaggatctcc ttttgggtgg
     5101 aaggacatca aggaatggta cgagatgctt atgggacatt gtacatattt tccggaggag
     5161 ctcaggagcg tcaagtacgc ctacaatgcc gacctgtaca atgccctcaa tgacctcaat
     5221 aacctcgtga ttaccaggga cgagaacgag aagctggagt actatgaaaa gttccagatt
     5281 atcgagaatg tgtttaagca gaagaagaag ccgacactta agcagattgc aaaggaaatc
     5341 ctcgtgaatg aggaagatat caagggatac agagtgacaa gtacaggcaa gcccgagttc
     5401 acaaatctga aggtgtacca cgatattaag gacataaccg cacgaaagga gataatcgaa
     5461 aacgctgagc tcctcgatca gatcgcaaaa attcttacca tctaccagtc tagtgaggac
     5521 attcaggagg aactgactaa tctgaacagt gagctcaccc aagaggaaat tgagcagatt
     5581 tcaaacctga aaggctacac cgggacgcac aatctgagcc tcaaagcaat caacctcatt
     5641 ctggatgaac tttggcacac aaatgacaac caaattgcca tattcaaccg cctgaaactg
     5701 gtgccaaaaa aagtggatct gtcacagcaa aaggaaatcc ctacaacctt ggttgacgat
     5761 tttattctgt cccccgttgt caagcggagc ttcatccagt caatcaaggt gatcaatgcc
     5821 atcattaaaa aatacggatt gccaaacgat ataattatcg agcttgcacg agagaagaac
     5881 tcaaaggacg cccagaagat gattaacgaa atgcagaagc gcaaccgcca gacaaacgaa
     5941 cgcatagagg aaattataag aacaaccggc aaagagaatg ccaagtatct gatcgagaaa
     6001 atcaagctgc acgacatgca agaaggcaag tgcctgtact ctctggaagc tatcccactc
     6061 gaagatctgc tgaataatcc attcaattac gaggtggacc acatcatccc tagatccgta
     6121 agctttgaca attccttcaa taacaaagtt ctggttaaac aggaggaaaa ttctaaaaaa
     6181 gggaaccgga ccccgttcca gtacctgagc tccagtgaca gcaagattag ctacgagact
     6241 tttaagaaac atattctgaa tctggccaaa ggcaaaggca ggatcagcaa gaccaagaag
     6301 gagtacctcc tcgaagaacg cgacattaac agatttagtg tgcagaaaga tttcatcaac
     6361 cgaaaccttg tcgatactcg gtacgccacg agaggcctga tgaatctcct caggagctac
     6421 ttccgcgtca ataatctgga cgttaaagtc aagagcataa atgggggatt caccagcttt
     6481 ctgaggagaa agtggaagtt taagaaggaa cgaaacaaag gatacaagca ccatgctgag
     6541 gatgctttga tcatcgctaa cgcggacttt atctttaagg aatggaaaaa gctggataag
     6601 gcaaagaaag tgatggaaaa ccagatgttc gaggagaagc aggcagagtc aatgcctgag
     6661 atcgagacag agcaggaata caaggaaatt ttcatcaccc ctcatcagat taaacacata
     6721 aaggacttca aagactataa atactctcat agggtggaca aaaaacccaa tcgcgagctc
     6781 attaatgaca ccctgtactc aacacggaag gatgataaag gtaatacctt gattgtgaat
     6841 aatcttaatg gattgtatga caaagataac gacaagctca agaagctgat caacaagtct
     6901 ccagagaagc tccttatgta tcaccacgac ccacagactt atcagaaatt gaaactgatc
     6961 atggagcaat acggggatga gaagaaccca ctctacaaat attatgagga aacaggtaat
     7021 tacctgacca agtactccaa gaaggataac ggaccagtga tcaaaaagat aaagtactat
     7081 ggcaacaaac ttaatgcgca tttggacata actgacgatt accccaattc tcgaaacaag
     7141 gttgtgaagc tctccctgaa gccttataga tttgacgtgt acctggataa tggggtttat
     7201 aaattcgtca ccgtgaaaaa tctggacgtg atcaaaaagg agaactatta tgaagtaaac
     7261 tcaaagtgct atgaggaggc gaagaagctg aagaagatct ccaatcaggc cgagttcatc
     7321 gcttccttct ataataacga tctcatcaag atcaatggag agctttatcg cgtcattggt
     7381 gtgaacaatg acttgctgaa caggatcgaa gtcaatatga tagacattac ctaccgggag
     7441 tatctcgaaa acatgaatga taaacggccg cctcgcatca tcaagacaat cgcatctaaa
     7501 actcagtcaa taaaaaagta ctctaccgat atcctgggga atctctatga agtgaagtca
     7561 aagaagcacc cacaaatcat taaaaaaggt ggatccccca agaagaagag gaaagtctcg
     7621 agcgactaca aagaccatga cggtgattat aaagatcatg acatcgatta caaggatgac
     7681 gatgacaag
//
