LOCUS       sequence-189524-        8256 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SaBE3-Gam in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8256)
  AUTHORS   Komor AC, Zhao KT, Packer MS, Gaudelli NM, Waterbury AL, Koblan LW, 
            Kim YB, Badran AH, Liu DR
  TITLE     Improved base excision repair inhibition and bacteriophage Mu Gam 
            protein yields C:G-to-T:A base editors with higher efficiency and 
            product purity.
  JOURNAL   Sci Adv. 2017 Aug 30;3(8):eaao4774. doi: 10.1126/sciadv.aao4774. 
            eCollection 2017 Aug.
  PUBMED    28875174
REFERENCE   2  (bases 1 to 8256)
  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: "Sci Adv."; 
            date: "2017-08-30"; volume: "3(8):eaao4774. doi"; pages: " 
            10.1126/sciadv.aao4774. eCollection 2017 Aug"
COMMENT     Sequence Label: SaBE3-Gam
FEATURES             Location/Qualifiers
     source          1..8256
                     /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"
     regulatory      3691..3700
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4267..4950
                     /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             8164..8184
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8191..8256
                     /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 gccaccatgg ctaaaccagc aaaacgtatc
     3721 aagagtgccg cagcggctta tgtgccacaa aaccgcgatg cggtgattac cgatattaaa
     3781 cgcatcgggg atttacagcg cgaagcatca cgtctggaaa cggaaatgaa tgatgccatc
     3841 gcggaaatta cggagaaatt tgcggcccgg attgcaccga ttaaaaccga tattgaaacc
     3901 ctttcaaaag gcgttcaggg atggtgtgaa gcgaaccgcg acgaactgac gaacggcggc
     3961 aaagtgaaga cggcgaatct tgtcaccggt gatgtatcgt ggcgggtccg tccaccatca
     4021 gtaagtattc gtggtatgga tgcagtgatg gaaacgctgg agcgtcttgg cctgcaacgc
     4081 tttattcgca cgaagcagga aatcaacaag gaagcgattt tactggaacc gaaagcggtc
     4141 gcaggcgttg ccggaattac agttaaatca ggcattgagg atttttctat tattccattt
     4201 gaacaggaag ccggtattag cggcagcgag actcccggga cctcagagtc cgctacaccc
     4261 gaaagtagct cagagactgg cccagtggct gtggacccca cattgagacg gcggatcgag
     4321 ccccatgagt ttgaggtatt cttcgatccg agagagctcc gcaaggagac ctgcctgctt
     4381 tacgaaatta attggggggg ccggcactcc atttggcgac atacatcaca gaacactaac
     4441 aagcacgtcg aagtcaactt catcgagaag ttcacgacag aaagatattt ctgtccgaac
     4501 acaaggtgca gcattacctg gtttctcagc tggagcccat gcggcgaatg tagtagggcc
     4561 atcactgaat tcctgtcaag gtatccccac gtcactctgt ttatttacat cgcaaggctg
     4621 taccaccacg ctgacccccg caatcgacaa ggcctgcggg atttgatctc ttcaggtgtg
     4681 actatccaaa ttatgactga gcaggagtca ggatactgct ggagaaactt tgtgaattat
     4741 agcccgagta atgaagccca ctggcctagg tatccccatc tgtgggtacg actgtacgtt
     4801 cttgaactgt actgcatcat actgggcctg cctccttgtc tcaacattct gagaaggaag
     4861 cagccacagc tgacattctt taccatcgct cttcagtctt gtcattacca gcgactgccc
     4921 ccacacattc tctgggccac cgggttgaaa agcggcagcg agactcccgg gacctcagag
     4981 tccgccacac ccgaaagtgg gaaacggaac tacatcctgg ggcttgccat tgggataacc
     5041 agcgttggct acggaattat tgattatgag acacgcgatg tgattgacgc cggggttagg
     5101 ctgttcaaag aggccaacgt tgaaaacaac gagggaagac ggagtaagcg cggagcaaga
     5161 agactcaagc gcagacggag acatcggatt cagagggtga aaaagctgct cttcgattac
     5221 aatctcctga ccgatcatag tgagctgagc ggaatcaacc cctacgaggc gcgagtgaaa
     5281 gggctttccc agaagctgtc cgaagaggag ttctccgccg cgttgctgca cctggccaaa
     5341 cggagggggg ttcacaatgt aaacgaagtg gaggaggaca cgggcaatga acttagtacg
     5401 aaagaacaga tcagtaggaa ctctaaggct ctcgaagaga aatacgtcgc tgagttgcag
     5461 cttgagagac tgaaaaaaga cggcgaagta cgcggatcta ttaataggtt caagacttca
     5521 gattacgtaa aggaagccaa gcagctcctg aaagtacaga aagcgtacca tcagctcgat
     5581 cagagcttca tcgataccta catagatttg ctggagacac ggaggacata ctacgagggc
     5641 ccaggggaag gatctccttt tgggtggaag gacatcaagg aatggtacga gatgcttatg
     5701 ggacattgta catattttcc ggaggagctc aggagcgtca agtacgccta caatgccgac
     5761 ctgtacaatg ccctcaatga cctcaataac ctcgtgatta ccagggacga gaacgagaag
     5821 ctggagtact atgaaaagtt ccagattatc gagaatgtgt ttaagcagaa gaagaagccg
     5881 acacttaagc agattgcaaa ggaaatcctc gtgaatgagg aagatatcaa gggatacaga
     5941 gtgacaagta caggcaagcc cgagttcaca aatctgaagg tgtaccacga tattaaggac
     6001 ataaccgcac gaaaggagat aatcgaaaac gctgagctcc tcgatcagat cgcaaaaatt
     6061 cttaccatct accagtctag tgaggacatt caggaggaac tgactaatct gaacagtgag
     6121 ctcacccaag aggaaattga gcagatttca aacctgaaag gctacaccgg gacgcacaat
     6181 ctgagcctca aagcaatcaa cctcattctg gatgaacttt ggcacacaaa tgacaaccaa
     6241 attgccatat tcaaccgcct gaaactggtg ccaaaaaaag tggatctgtc acagcaaaag
     6301 gaaatcccta caaccttggt tgacgatttt attctgtccc ccgttgtcaa gcggagcttc
     6361 atccagtcaa tcaaggtgat caatgccatc attaaaaaat acggattgcc aaacgatata
     6421 attatcgagc ttgcacgaga gaagaactca aaggacgccc agaagatgat taacgaaatg
     6481 cagaagcgca accgccagac aaacgaacgc atagaggaaa ttataagaac aaccggcaaa
     6541 gagaatgcca agtatctgat cgagaaaatc aagctgcacg acatgcaaga aggcaagtgc
     6601 ctgtactctc tggaagctat cccactcgaa gatctgctga ataatccatt caattacgag
     6661 gtggaccaca tcatccctag atccgtaagc tttgacaatt ccttcaataa caaagttctg
     6721 gttaaacagg aggaaaattc taaaaaaggg aaccggaccc cgttccagta cctgagctcc
     6781 agtgacagca agattagcta cgagactttt aagaaacata ttctgaatct ggccaaaggc
     6841 aaaggcagga tcagcaagac caagaaggag tacctcctcg aagaacgcga cattaacaga
     6901 tttagtgtgc agaaagattt catcaaccga aaccttgtcg atactcggta cgccacgaga
     6961 ggcctgatga atctcctcag gagctacttc cgcgtcaata atctggacgt taaagtcaag
     7021 agcataaatg ggggattcac cagctttctg aggagaaagt ggaagtttaa gaaggaacga
     7081 aacaaaggat acaagcacca tgctgaggat gctttgatca tcgctaacgc ggactttatc
     7141 tttaaggaat ggaaaaagct ggataaggca aagaaagtga tggaaaacca gatgttcgag
     7201 gagaagcagg cagagtcaat gcctgagatc gagacagagc aggaatacaa ggaaattttc
     7261 atcacccctc atcagattaa acacataaag gacttcaaag actataaata ctctcatagg
     7321 gtggacaaaa aacccaatcg cgagctcatt aatgacaccc tgtactcaac acggaaggat
     7381 gataaaggta ataccttgat tgtgaataat cttaatggat tgtatgacaa agataacgac
     7441 aagctcaaga agctgatcaa caagtctcca gagaagctcc ttatgtatca ccacgaccca
     7501 cagacttatc agaaattgaa actgatcatg gagcaatacg gggatgagaa gaacccactc
     7561 tacaaatatt atgaggaaac aggtaattac ctgaccaagt actccaagaa ggataacgga
     7621 ccagtgatca aaaagataaa gtactatggc aacaaactta atgcgcattt ggacataact
     7681 gacgattacc ccaattctcg aaacaaggtt gtgaagctct ccctgaagcc ttatagattt
     7741 gacgtgtacc tggataatgg ggtttataaa ttcgtcaccg tgaaaaatct ggacgtgatc
     7801 aaaaaggaga actattatga agtaaactca aagtgctatg aggaggcgaa gaagctgaag
     7861 aagatctcca atcaggccga gttcatcgct tccttctata ataacgatct catcaagatc
     7921 aatggagagc tttatcgcgt cattggtgtg aacaatgact tgctgaacag gatcgaagtc
     7981 aatatgatag acattaccta ccgggagtat ctcgaaaaca tgaatgataa acggccgcct
     8041 cgcatcatca agacaatcgc atctaaaact cagtcaataa aaaagtactc taccgatatc
     8101 ctggggaatc tctatgaagt gaagtcaaag aagcacccac aaatcattaa aaaaggtgga
     8161 tcccccaaga agaagaggaa agtctcgagc gactacaaag accatgacgg tgattataaa
     8221 gatcatgaca tcgattacaa ggatgacgat gacaag
//
