LOCUS       sequence-460120-        5148 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of apt-CBE-SL3 protein 
            (MCP_16aa-linker_evoAPOBEC1) with P2A-EGFP marker .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5148)
  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 5148)
  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: apt-CBE-SL3
FEATURES             Location/Qualifiers
     source          1..5148
                     /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             1813..2061
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             2092..2340
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             2383..2403
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2412..2429
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2452..2469)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2458..2682
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2753..2769)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2753..2769)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2766..2788)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2777..2793
                     /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(2801..2807,2808..2825,2826..2831))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2846..2867
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2984..3001)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3155..3743)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3235..3254)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3914..4705,4706..4774))
                     /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     4537..4556
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4775..4879)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4958..4977)
                     /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 tttagcagta 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 atccagtggc
     1081 ggcagcagcg gcggcagcag ttcaaagact gggcctgtcg ccgtcgatcc aaccctgcgc
     1141 cgccggattg aacctcacga gtttgaagtg ttctttgacc cccgggagct gagaaaggag
     1201 acatgcctgc tgtacgagat caactgggga ggcaggcact ccatctggag gcacacctct
     1261 cagaacacaa ataagcacgt ggaggtgaac ttcatcgaga agtttaccac agagcggtac
     1321 ttctgcccca ataccagatg tagcatcaca tggtttctga gctggtcccc ttgcggagag
     1381 tgtagcaggg ccatcaccga gttcctgtcc agatatccaa atgtgacact gtttatctac
     1441 atcgccaggc tgtatcacct ggcaaaccca aggaataggc agggcctgcg cgatctgatc
     1501 agctccggcg tgaccatcca gatcatgaca gagcaggagt ccggctactg ctggcacaac
     1561 ttcgtgaatt attctcctag caacgagtcc cactggccta ggtacccaca cctgtgggtg
     1621 cgcctgtacg tgctggagct gtattgcatc atcctgggcc tgcccccttg tctgaatatc
     1681 ctgcggagaa agcagagcca gctgacctcc tttacaatcg ccctgcagtc ttgtcactat
     1741 cagaggctgc caccccacat cctgtgggcc acaggcctga agtctggcgg atctggaggc
     1801 tccggcggca gcactaatct gagcgacatc attgaaaagg agactgggaa acagctggtc
     1861 attcaggagt ccatcctgat gctgcctgag gaggtggagg aagtgatcgg caacaagcca
     1921 gagtctgaca tcctggtgca caccgcctac gacgagtcca cagatgagaa tgtgatgctg
     1981 ctgacctctg acgcccccga gtataagcct tgggccctgg tcatccagga ttctaacggc
     2041 gagaataaga tcaagatgct gagcggagga tccggaggat ctggaggcag caccaacctg
     2101 tctgacatca tcgagaagga gacaggcaag cagctggtca tccaggagag catcctgatg
     2161 ctgcccgaag aagtcgaaga agtgatcgga aacaagcctg agagcgatat cctggtccat
     2221 accgcctacg acgagagtac cgacgaaaat gtgatgctgc tgacatccga cgccccagag
     2281 tataagccct gggctctggt catccaggat tccaacggag agaacaaaat caaaatgctg
     2341 tctggcggct caaaaagaac cgccgacggc agcgaattcg agcccaagaa gaaaaggaaa
     2401 gtctaaccgg tcatcatcac catcaccatt gagtttaaac ccgctgatca gcctcgactg
     2461 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     2521 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     2581 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     2641 aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa
     2701 ccagctgggg ctcgataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     2761 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     2821 taaagtgtaa agcctaggat gcctaatgag tgagctaact cacattaatt gcgttgcgct
     2881 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     2941 gcgcgggaag aagcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     3001 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     3061 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     3121 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     3181 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     3241 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     3301 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     3361 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     3421 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     3481 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     3541 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     3601 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     3661 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     3721 cgcgcagaaa aaaaggatct caagaaaatc ctttgatctt ttctacgggg tctgacactc
     3781 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     3841 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     3901 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     3961 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     4021 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     4081 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     4141 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     4201 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     4261 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     4321 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     4381 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     4441 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     4501 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     4561 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     4621 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     4681 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaaag
     4741 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     4801 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     4861 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg
     4921 gagatcgatc tcccgatccc ctagggtcga ctctcagtac aatctgctct gatgccgcat
     4981 agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca
     5041 aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttaggg
     5101 ttaggcgttt tgcgctgctt cgcgatgtac gggccagata tacgcgtt
//
