LOCUS       sequence-460115-        6285 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of MOBE2 protein 
            (apt-CBE-3'end_PT2A_apt-ABE8.20).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6285)
  AUTHORS   Cowan QT, Gu S, Gu W, Ranzau BL, Simonson TS, Komor AC
  TITLE     Development of multiplexed orthogonal base editor (MOBE) systems.
  JOURNAL   Nat Biotechnol. 2024 May 21. doi: 10.1038/s41587-024-02240-0.
  PUBMED    38773305
REFERENCE   2  (bases 1 to 6285)
  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. 2024 May 21. doi: 10.1038/s41587-024-02240-0."
COMMENT     Sequence Label: MOBE2
FEATURES             Location/Qualifiers
     source          1..6285
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      652..661
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             661..1047
                     /codon_start=1
                     /product="bacteriophage MS2 coat protein"
                     /label=MS2-N55K
                     /note="binds with high affinity to a specific stem-loop 
                     structure in the viral RNA (Lim et al., 1994)"
                     /translation="ASNFTQFVLVDNGGTGDVTVAPSNFANGVAEWISSNSRSQAYKVT
                     CSVRQSSAQKRKYTIKVEVPKVATQTVGGVELPVAAWRSYLNMELTIPIFATNSDCELI
                     VKAMQGLLKDGNPIPSAIAANSGIY"
     CDS             1927..2175
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             2206..2454
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             2497..2517
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2527..2583
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             2593..2646
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             2683..2703
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2707..3204
                     /codon_start=1
                     /product="DNA adenosine deaminase evolved from the E. coli 
                     tRNA adenosine deaminase TadA (Gaudelli et al., 2017)"
                     /label=TadA*(7.10)
                     /translation="SEVEFSHEYWMRHALTLAKRARDEREVPVGAVLVLNNRVIGEGWN
                     RAIGLHDPTAHAEIMALRQGGLVMQNYRLYDATLYSTFEPCVMCAGAMIHSRIGRVVFG
                     VRNAKTGAAGSLMDVLHHPGMNHRVEITEGILADECAALLCRFFRMPRRVFNAQKKAQS
                     STD"
     CDS             3520..3540
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3549..3566
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3589..3606)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3595..3819
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3890..3906)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3890..3906)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3903..3925)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3914..3930
                     /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(3938..3944,3945..3962,3963..3968))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3983..4004
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4121..4138)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4292..4880)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4372..4391)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5051..5842,5843..5911))
                     /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     5674..5693
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5912..6016)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6095..6114)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc
      601 agatccgcta gagatccgcg gccgctaata cgactcacta tagggagagc cgccaccatg
      661 gcaagtaact tcactcaatt tgtgcttgta gacaacggag gtaccggaga tgtaacagta
      721 gcacccagta atttcgcaaa cggcgtggcg gaatggatta gttcaaattc aagatctcag
      781 gcctataagg tcacttgctc cgtcaggcag tctagtgccc aaaagcgcaa atacacgatt
      841 aaagtagagg taccgaaggt cgctacgcaa actgttgggg gagttgaact tccagtggct
      901 gcttggcgct catacctgaa catggaactg actatcccca tattcgctac taactccgat
      961 tgtgaactca tagtaaaagc tatgcaggga ttgctcaagg atggcaaccc cattccaagt
     1021 gccatagcag cgaactccgg gatatattct tctggaggat ctagcggagg atcctctggc
     1081 agcgagacac caggaacaag cgagtcagca acaccagaga gcagtggcgg cagcagcggc
     1141 ggcagcagtt caaagactgg gcctgtcgcc gtcgatccaa ccctgcgccg ccggattgaa
     1201 cctcacgagt ttgaagtgtt ctttgacccc cgggagctga gaaaggagac atgcctgctg
     1261 tacgagatca actggggagg caggcactcc atctggaggc acacctctca gaacacaaat
     1321 aagcacgtgg aggtgaactt catcgagaag tttaccacag agcggtactt ctgccccaat
     1381 accagatgta gcatcacatg gtttctgagc tggtcccctt gcggagagtg tagcagggcc
     1441 atcaccgagt tcctgtccag atatccaaat gtgacactgt ttatctacat cgccaggctg
     1501 tatcacctgg caaacccaag gaataggcag ggcctgcgcg atctgatcag ctccggcgtg
     1561 accatccaga tcatgacaga gcaggagtcc ggctactgct ggcacaactt cgtgaattat
     1621 tctcctagca acgagtccca ctggcctagg tacccacacc tgtgggtgcg cctgtacgtg
     1681 ctggagctgt attgcatcat cctgggcctg cccccttgtc tgaatatcct gcggagaaag
     1741 cagagccagc tgacctcctt tacaatcgcc ctgcagtctt gtcactatca gaggctgcca
     1801 ccccacatcc tgtgggccac aggcctgaag tctggcggat ctagcggagg atcctctggc
     1861 agcgagacac caggaacaag cgagtcagca acaccagaga gcagtggcgg cagcagcggc
     1921 ggcagcacta atctgagcga catcattgag aaggagactg ggaaacagct ggtcattcag
     1981 gagtccatcc tgatgctgcc tgaggaggtg gaggaagtga tcggcaacaa gccagagtct
     2041 gacatcctgg tgcacaccgc ctacgacgag tccacagatg agaatgtgat gctgctgacc
     2101 tctgacgccc ccgagtataa gccttgggcc ctggtcatcc aggattctaa cggcgagaat
     2161 aagatcaaga tgctgagcgg aggatccgga ggatctggag gcagcaccaa cctgtctgac
     2221 atcatcgaga aggagacagg caagcagctg gtcatccagg agagcatcct gatgctgccc
     2281 gaagaagtcg aagaagtgat cggaaacaag cctgagagcg atatcctggt ccataccgcc
     2341 tacgacgaga gtaccgacga aaatgtgatg ctgctgacat ccgacgcccc agagtataag
     2401 ccctgggctc tggtcatcca ggattccaac ggagagaaca aaatcaaaat gctgtctggc
     2461 ggctcaaaaa gaaccgccga cggcagcgaa ttcgagccca agaagaagag gaaagtcgga
     2521 agcggagcta ctaacttcag cctgctgaag caggctggag acgtggagga gaaccctgga
     2581 cctggcagtg gtgagggcag aggaagtctg ctaacatgcg gtgacgtcga ggagaatcct
     2641 ggcccaatga aacggacagc cgacggaagc gagttcgagt caccaaagaa gaagcggaaa
     2701 gtcagctctg aggtggagtt ttcccatgag tactggatga gacacgcatt gactctcgca
     2761 aagagggctc gagatgaacg cgaggtgccc gtgggggcag tactcgtgct caacaatcgc
     2821 gtaatcggcg aaggttggaa tagggcaatc ggactccacg accccactgc acatgcggaa
     2881 atcatggccc ttcgacaggg agggcttgtg atgcagaatt atcgacttta tgatgcgacg
     2941 ctgtactcga cgtttgaacc ttgcgtaatg tgcgcgggag ctatgattca ctcccgcatt
     3001 ggacgagttg tattcggtgt tcgcaacgcc aagacgggtg ccgcaggttc actgatggac
     3061 gtgctgcatc atccaggcat gaaccaccgg gtagaaatca cagaaggcat attggcggac
     3121 gaatgtgcgg cgctgttgtg tcgttttttt cgcatgccca ggcgggtctt taacgcccag
     3181 aaaaaagcac aatcctctac tgactctggt ggttcttctg gtggttctag cggcagcgag
     3241 actcccggga cctcagagtc cgccacaccc gaaagttctg gtggttcttc tggtggttct
     3301 aaaagcatca gatgcaagaa ctgcaacaag ctgctgttca aggccgacag cttcgaccac
     3361 atcgagatcc ggtgtcctag atgtaaaaga cacatcatca tgctgaacgc ctgcgagcac
     3421 cccaccgaga agcactgcgg caagcgggaa aagatcaccc acagcgacga gacagtgcgg
     3481 tacggttcaa aaagaactgc cgacggctct gagtttgaac ccaagaagaa aaggaaagtc
     3541 taaccggtca tcatcaccat caccattgag tttaaacccg ctgatcagcc tcgactgtgc
     3601 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     3661 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     3721 ggtgtcattc tattctgggg ggtggggtga ggcaggacag caagggggag gattgggaag
     3781 acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca
     3841 gctggggctc gataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     3901 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     3961 agtgtaaagc ctaggatgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     4021 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     4081 cgggaaaagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     4141 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     4201 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     4261 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     4321 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     4381 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     4441 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     4501 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     4561 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     4621 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     4681 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     4741 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     4801 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     4861 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacactcagt
     4921 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     4981 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     5041 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     5101 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     5161 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     5221 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     5281 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     5341 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     5401 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     5461 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     5521 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     5581 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     5641 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     5701 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     5761 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     5821 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     5881 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     5941 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     6001 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag
     6061 atcgatctcc cgatccccta gggtcgactc tcagtacaat ctgctctgat gccgcatagt
     6121 taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa
     6181 tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagggtta
     6241 ggcgttttgc gctgcttcgc gatgtacggg ccagatatac gcgtt
//
