LOCUS       sequence-373432-        6885 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Hypercompact adenine base editors.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6885)
  AUTHORS   Kim DY, Chung Y, Lee Y, Jeong D, Park KH, Chin HJ, Lee JM, Park S, 
            Ko S, Ko JH, Kim YS
  TITLE     Hypercompact adenine base editors based on transposase B guided by 
            engineered RNA.
  JOURNAL   Nat Chem Biol. 2022 Sep;18(9):1005-1013. doi: 
            10.1038/s41589-022-01077-5. Epub 2022 Aug 1.
  PUBMED    35915259
REFERENCE   2  (bases 1 to 6885)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/s41589-022-01077-5"; journalName: "Nat Chem Biol"; date: 
            "2022-09"; volume: "18"; issue: "9"; pages: "1005-1013"
COMMENT     Sequence Label: TaRGET-ABE-C3.1 ver4.1
FEATURES             Location/Qualifiers
     source          1..6885
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        118..497
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        498..701
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     652..672
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      743..752
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             755..775
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2567..3064
                     /codon_start=1
                     /product="tRNA adenosine deaminase from E. coli"
                     /label=TadA
                     /translation="SEVEFSHEYWMRHALTLAKRAWDEREVPVGAVLVHNNRVIGEGWN
                     RPIGRHDPTAHAEIMALRQGGLVMQNYRLIDATLYVTLEPCVMCAGAMIHSRIGRVVFG
                     ARDAKTGAAGSLMDVLHHPGMNHRVEITEGILADECAALLSDFFRMRRQEIKAQKKAQS
                     STD"
     CDS             3659..3706
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             3719..3745
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             3719..3745
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(3768..3785)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3774..3998
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4069..4085)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4069..4085)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4082..4104)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4093..4109
                     /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(4117..4123,4124..4141,4142..4147))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4162..4183
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4300..4317)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4471..5059)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4551..4570)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5230..6021,6022..6090))
                     /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     5853..5872
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6091..6195)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6274..6293)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        6464..6712
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     6464..6484
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     6635..6654
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
ORIGIN
        1 cggatcccga agacgatttt attttttgtt ttagagctag aaatagtaag ttaaaataag
       61 gctagtccgt ttttagcgcg tgcgccaatt ctgcagacaa atggctctag aggtaccgac
      121 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      181 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      241 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      301 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      361 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      421 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      481 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      541 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      601 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      661 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgtcaga
      721 tccgctagag atccgcggcc gcgccaccat ggccccaaag aagaagcgga aggtcggtat
      781 ccacggagtc ccagcagcca tgggggagaa aagttcccgc cgccgacgga atggaaaaag
      841 cggtgcgtgg actgctgcta taacaagctg tgttgggggt aagatggcca agaacacaat
      901 tacaaagaca ctgaagctga ggatcgtgag accatacaac agcgctgagg tcgagaagat
      961 tgtggctgat gaaaagaaca acagggaaaa gatcgccctc gagaagaaca aggataaggt
     1021 gaaggaggcc tgctctaagc acctgaaagt ggccgcctac tgcaccacac aggtggagag
     1081 gaacgcctgt ctgttttgta aagctcggaa gctggatgat aagttttacc agaagctgcg
     1141 gggccagttc cccgatgccg tcttttggca ggagattagc gagatcttca gacagctgca
     1201 gaagcaggcc gccgagatct acaaccagag cctgatcgag ctctactacg agatcttcat
     1261 caagggcaag ggcattgcca acgcctcctc cgtggagcac tacctgagcg acgtgtgcta
     1321 cacaagagcc gccgagctct ttaagaacgc cgctatcgct tccgggctga ggagcaagat
     1381 taagagtaac ttccggctca aggagctgaa gaacatgaag agcggcctgc ccactacaaa
     1441 gagcgacaac ttcccaattc cactggtgaa gcagaagggg ggccagtaca cagggttcga
     1501 gatttccaac cacaacagcg actttattat taagatcccc tttggcaggt ggcaggtcaa
     1561 gaaggagatt gacaagtaca ggccctggga gaagtttgat ttcgagcagg tgcagaagag
     1621 ccccaagcct atttccctgc tgctgtccac acagcggcgg aagaggaaca aggggtggtc
     1681 taaggatgag gggaccgagg ccgagattaa gaaagtgatg aacggcgact accagacaag
     1741 ctacatcgag gtcaagcggg gcagtaagat tggcgagaag agcgcctgga tgctgaacct
     1801 gagcattgac gtgccaaaga ttgataaggg cgtggatccc agcatcatcg gagggatcgc
     1861 tgtgggggtc aagagccccc tcgtgtgcgc catcaacaac gccttcagca ggtacagcat
     1921 ctccgataac gacctgttcc actttaacaa gaagatgttc gcccggcgga ggattttgct
     1981 caagaagaac cggcacaagc gggccggaca cggggccaag aacaagctca agcccatcac
     2041 tatcctgacc gagaagagcg agaggttcag gaagaagctc atcgagagat gggcctgcga
     2101 gatcgccgat ttctttatta agaacaaggt cggaacagtg cagatggaga acctcgagag
     2161 catgaagagg aaggaggatt cctacttcaa cattcggctg agggggttct ggccctacgc
     2221 tgagatgcag aacaagattg agtttaagct gaagcagtac gggattgaga tccggaaggt
     2281 ggcccccaac aacaccagca agacctgcag caagtgcggg cacctcaaca actacttcaa
     2341 cttcgagtac cggaagaaga acaagttccc acacttcaag tgcgagaagt gcaactttaa
     2401 ggagaacgcc gattacaacg ccgccctgaa catcagcaac cctaagctga agagcactaa
     2461 ggaggagccc tctggaggat ctagcggagg atcctctggc agcgagacac caggaacaag
     2521 cgagtcagca acaccagaga gcagtggcgg cagcagcggc ggcagctccg aagtcgagtt
     2581 ttcccatgag tactggatga gacacgcatt gactctcgca aagagggctt gggatgaacg
     2641 cgaggtgccc gtgggggcag tactcgtgca taacaatcgc gtaatcggcg aaggttggaa
     2701 taggccgatc ggacgccacg accccactgc acatgcggaa atcatggccc ttcgacaggg
     2761 agggcttgtg atgcagaatt atcgacttat cgatgcgacg ctgtacgtca cgcttgaacc
     2821 ttgcgtaatg tgcgcgggag ctatgattca ctcccgcatt ggacgagttg tattcggtgc
     2881 ccgcgacgcc aagacgggtg ccgcaggttc actgatggac gtgctgcatc acccaggcat
     2941 gaaccaccgg gtagaaatca cagaaggcat attggcggac gaatgtgcgg cgctgttgtc
     3001 cgactttttt cgcatgcgga ggcaggagat caaggcccag aaaaaagcac aatcctctac
     3061 tgactctggt ggttcttctg gtggttctag cggcagcgag actcccggga cctcagagtc
     3121 cgccacaccc gaaagttcag gtggatcttc aggtggatct tctgaggtgg agttttccca
     3181 cgagtactgg atgagacatg ccctgaccct ggccaagagg gcacgggatg agagggaggt
     3241 gcctgtggga gccgtgctgg tgctgaacaa tagagtgatc ggcgagggct ggaacagagc
     3301 catcggcctg cacgacccaa cagcccatgc cgaaattatg gccctgagac agggcggcct
     3361 ggtcatgcag aactacagac tgattgacgc caccctgtac gtgacattcg agccttgcgt
     3421 gatgtgcgcc ggcgccatga tccactctag gatcggccgc gtggtgtttg gatggagaca
     3481 gtctaaaaga ggcgccgcag gctccctgat gaacgtgctg aactaccccg gcatgaatca
     3541 ccgcgtcgaa attaccgagg gaatcctggc agatgaatgt gccgccctgc tgtgcgattt
     3601 ctatcggatg cctagacagg tgttcaatgc tcagaagaag gcccagagct ccatcaacaa
     3661 aaggccggcg gccacgaaaa aggccggcca ggcaaaaaag aaaaagtctg gtggttctta
     3721 cccctacgac gtgcccgact acgcctaagt ttaaacccgc tgatcagcct cgactgtgcc
     3781 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     3841 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     3901 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     3961 gaatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     4021 ctggggctcg ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     4081 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     4141 gtgtaaagcc tagggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     4201 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     4261 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     4321 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     4381 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     4441 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     4501 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     4561 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     4621 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     4681 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     4741 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     4801 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     4861 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     4921 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     4981 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     5041 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     5101 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     5161 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     5221 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     5281 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     5341 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     5401 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     5461 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     5521 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     5581 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     5641 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     5701 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     5761 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     5821 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     5881 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     5941 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     6001 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     6061 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     6121 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     6181 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga
     6241 tcgatctccc gatcccctag ggtcgactct cagtacaatc tgctctgatg ccgcatagtt
     6301 aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat
     6361 ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagggttag
     6421 gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgtgagggcc tatttcccat
     6481 gattccttca tatttgcata tacgatacaa ggctgttaga gagataattg gaattaattt
     6541 gactgtaaac acaaagatat tagtacaaaa tacgtgacgt agaaagtaat aatttcttgg
     6601 gtagtttgca gttttaaaat tatgttttaa aatggactat catatgctta ccgtaacttg
     6661 aaagtatttc gatttcttgg ctttatatat cttgtggaaa ggacgaaaca ccgaccgctt
     6721 cacttagagt gaaggtgggc tgcttgcatc agcctaatgt cgagaagtgc tttcttcgga
     6781 aagtaaccct cgaaacaaag aaaggaatgc aacgagtctt cgacgcacct gacgtctaag
     6841 aaaccattat tatcatgaca ttaacctata aaaataggcg tatca
//
