LOCUS       sequence-442604-        7978 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Adenine base editor with modified TadA variant for A-to-G base 
            editing in zebrafish.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7978)
  AUTHORS   Qin W, Liang F, Lin SJ, Petree C, Huang K, Zhang Y, Li L, Varshney 
            P, Mourrain P, Liu Y, Varshney GK
  TITLE     ABE-ultramax for high-efficiency biallelic adenine base editing in 
            zebrafish.
  JOURNAL   Nat Commun. 2024 Jul 4;15(1):5613. doi: 10.1038/s41467-024-49943-1.
  PUBMED    38965236
REFERENCE   2  (bases 1 to 7978)
  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 
            Commun."; date: "2024-07-4"; volume: "15(1):5613. doi"; pages: " 
            10.1038/s41467-024-49943-1"
COMMENT     Sequence Label: pT3TS-spCas9 ABE-Umax
FEATURES             Location/Qualifiers
     source          1..7978
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    877..898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(913..918,919..936,937..943)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    951..967
                     /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)."
     primer_bind     956..978
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     975..991
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     975..991
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        1012..1030
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     5'UTR           1055..1097
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     CDS             1155..1175
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     RBS             5789..5797
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             5910..5930
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     3'UTR           5955..6080
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
     primer_bind     complement(6177..6196)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6178..6196)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6203..6220)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6203..6219)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6212..6234)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(6361..6816)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6498..6519)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6710..6729
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6843..6947
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6948..7016,7017..7808)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPLFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(7166..7185)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      961 ataacaattt cacacaggaa acagctatga ccatgattac gccaagctcg aaattaaccc
     1021 tcactaaagg gaacaaaagc tggagctaag cttgcttgtt ctttttgcag aagctcagaa
     1081 taaacgctca actttggcag atctaactcg acgccaccat gaaaagaact gcagatggat
     1141 ccgagttcga gagccccaag aaaaagagaa aggtgggaag catgagcgaa gtcgagtttt
     1201 cacacgaata ctggatgaga catgctctca cactcgcaaa gagggctaga gatgagaggg
     1261 aggttcctgt gggcgcagtg ctggtcctga ataacagggt tataggcgag ggatggaacc
     1321 gcgcaattgg actgcatgac ccaaccgcac acgcagagat tatggcactc aggcaaggag
     1381 gactggtcat gcagaactat agactcattg acgccacact gtatagcacc ttcgagcctt
     1441 gtgtgatgtg cgcaggagcc atgatccaca gtaggatcgg tagagtcgtg tttggagtta
     1501 gaaactcaaa gcgcggagct gccggcagtc tgatgaatgt tctgaactac ccaggaatga
     1561 atcatagggt cgagatcacc gaaggcattc tggccgacga atgtgccgct ctgctgtgcg
     1621 atttctacag aatgccaaga cgggttttca acgctcagaa gaaagcccag tcttccatta
     1681 atagcggagg ttccagcgga ggctccagtg gaagcgaaac acctgggaca tcagagtctg
     1741 ccactccaga gtccagtgga ggctcctctg gcggcagtga taagaagtat agcatcggcc
     1801 tggctattgg aactaactcc gtgggttggg cagtgattac agacgactac aaggtcccta
     1861 gcaagaaatt taaggtgctg ggtaacaccg acaggcacag catcaagaaa aatctgattg
     1921 gagccctgct gttcggttct ggagagactg ccgaagcaac acgcctgaaa agaacagcaa
     1981 gaaggcgcta taccagaagg aagaatagaa tctgttacct gcaggagatt ttctctaacg
     2041 aaatggctaa ggtggacgat tcattctttc ataggctgga ggaaagtttc ctggtcgagg
     2101 aagataagaa acacgagcgc catcctatct ttggaaacat tgtggacgag gtcgcctatc
     2161 acgaaaaata cccaaccatc tatcatctgc gcaagaaact ggctgactct actgataaag
     2221 ccgacctgag actgatctat ctggctctgg cccacatgat taagttcagg ggtcattttc
     2281 tgatcgaggg cgatctgaac cccgacaatt ccgatgtgga caagctgttc atccagctgg
     2341 tccagattta caatcagctg tttgaggaaa accctattaa tgcttccaga gtggacgcaa
     2401 aagctatcct gtcagccagg ctgtccaagt cacgcagact ggagaacctg attgcacagc
     2461 tgcccggaga aaagaggaac ggtctgtttg gaaatctgat cgctctgagt ctgggcctga
     2521 ctcctaactt caaaagcaat tttgatctgg ctgaggacgc caaactgcag ctgtcaaagg
     2581 acacatatga cgatgacctg gataacctgc tggcacagat cggagatcag tacgctgacc
     2641 tgttcctggc tgccaaaaat ctgtccgacg caatcctgct gtcagatatt ctgagagtga
     2701 acagcgagat tacaaaagct cctctgagtg ccagcatgat caagagatat gacgagcacc
     2761 atcaggatct gaccctgctg aaggctctgg tcaggcagca gctgccagag aagtacaagg
     2821 aaattttctt tgatcagtcc aagaacggct acgccggtta tatcgacgga ggcgcatcac
     2881 aggaggaatt ctacaagttt atcaaaccta ttctggagaa gatggacgga actgaggaac
     2941 tgctggtgaa actgaataga gaggacctgc tgaggaagca gcgcacattt gataacggtt
     3001 ccatcccaca ccagattcat ctgggagagc tgcacgctat cctgaggcgc caggaagact
     3061 tctacccctt tctgaaagat aaccgcgaga agatcgaaaa aattctgacc ttcagaatcc
     3121 cttactatgt gggtccactg gctcgcggaa acagcagatt tgcctggatg actcgcaaat
     3181 ccgaggaaac cattactcct tggaacttcg aggaagtggt cgataagggc gcctctgcac
     3241 agtccttcat cgagagaatg actaattttg acaaaaacct gcccaatgag aaagtgctgc
     3301 ctaagcactc cctgctgtac gagtatttca ctgtctataa cgaactgaca aaggtgaaat
     3361 acgtcaccga gggcatgaga aagccagcct tcctgtcagg agagcagaag aaagcaatcg
     3421 tggatctgct gtttaaaacc aacaggaaag tgactgtcaa gcagctgaag gaggactact
     3481 tcaagaaaat tgaatgcttc gattccgtgg agatcagcgg agtcgaagac agatttaacg
     3541 caagcctggg cgcttaccac gatctgctga agatcattaa ggataaagac ttcctggaca
     3601 acgaggaaaa tgaggatatc ctggaagaca ttgtgctgac actgaccctg tttgaggaca
     3661 gaggaatgat cgaggaaaga ctgaaaacct atgctcatct gttcgatgac aaggtgatga
     3721 aacagctgaa gagaaggcgc tacactggct ggggtagact gagcaggaag ctgatcaacg
     3781 gcattaggga taaacagtca ggaaagacaa tcctggactt tctgaaaagt gatggcttcg
     3841 ccaaccgcaa ttttatgcag ctgattcacg atgacagtct gaccttcaaa gaggacatcc
     3901 agaaggctca ggtgtctgga cagggccact ccctgcatga gcagattgca aacctggctg
     3961 gaagcccagc catcaagaaa ggcattctgc agacagtgaa aatcgtcgat gagctggtga
     4021 aagtcatggg ccataagccc gaaaacatcg tgattgagat ggctcgcgaa aatcagacaa
     4081 cccagaaggg tcagaagaac agtagagaga ggatgaaaag aatcgaggaa ggcattaagg
     4141 agctgggtag ccagatcctg aaagagcacc cagtggaaaa cacacagctg cagaatgaga
     4201 agctgtatct gtactatctg cagaatggaa gagatatgta cgtggaccag gagctggata
     4261 ttaacaggct gtctgattac gacgtggatc atatcgtccc ccagagtttc atcaaagatg
     4321 acagcattga caacaaggtg ctgaccaggt ccgacaaaaa cagaggaaaa tcagataatg
     4381 tccctagtga ggaagtggtc aagaaaatga agaactactg gagacagctg ctgaatgcca
     4441 aactgatcac tcagaggaag tttgataacc tgacaaaagc agagcgcggt ggactgtcag
     4501 aactggacaa agctggattc atcaagaggc agctggtgga aacacgccag atcactaaac
     4561 acgtcgcaca gattctggat agtcgcatga acacaaagta cgatgagaat gacaaactga
     4621 tcagagaagt gaaggtcatt accctgaaga gtaaactggt cagcgacttt aggaaagatt
     4681 tccagtttta taaggtccgc gagattaaca attatcacca tgcccatgac gcatacctga
     4741 acgccgtggt cggtaccgca ctgatcaaga aatacccaaa actggagagc gaattcgtgt
     4801 acggagacta taaggtgtac gatgtcagaa aaatgatcgc caagtccgag caggaaattg
     4861 gaaaagctac tgccaagtat ttcttttact caaacatcat gaatttcttt aagacagaga
     4921 tcaccctggc caatggagaa atccgcaaaa ggcccctgat tgagacaaac ggagagacag
     4981 gcgaaatcgt gtgggacaaa ggcagagatt ttgcaaccgt gaggaaggtc ctgagcatgc
     5041 ctcaagtgaa tatcgtcaag aaaactgagg tgcagacagg cggtttctca aaagaaagta
     5101 ttctgccaaa acgcaactct gataagctga tcgctagaaa gaaagactgg gaccctaaga
     5161 agtatggagg ctttgactct cccactgtgg catactccgt cctggtggtc gctaaggtgg
     5221 agaagggcaa aagcaagaaa ctgaaatctg tcaaggagct gctgggtatc acaattatgg
     5281 agagaagctc tttcgagaag aacccaatcg attttctgga ggccaaaggt tataaggaag
     5341 tgaagaaaga cctgatcatt aaactgccca agtacagtct gtttgagctg gaaaacggca
     5401 ggaaacgcat gctggcaagc gctggagagc tgcagaaagg caatgaactg gccctgcctt
     5461 ctaagtacgt gaacttcctg tatctggcaa gccactacga gaagctgaaa ggatctccag
     5521 aggataacga acagaaacag ctgtttgtgg agcagcacaa gcattatctg gacgagatca
     5581 ttgaacagat tagcgagttc tctaaaagag tgatcctggc cgacgcaaat ctggataagg
     5641 tcctgtctgc ttacaacaaa cacagagata agcccatcag ggagcaggcc gaaaatatca
     5701 ttcatctgtt cactctgaca aacctgggcg cacctgcagc tttcaagtac ttcgacacta
     5761 caatcgatag aaagaggtac acctccacta aggaggtgct ggacgctaca ctgatccatc
     5821 agagtattac cggcctgtac gaaacaagga ttgacctgtc tcagctgggt ggcgacaaaa
     5881 gaactgcaga tggatccgag ttcgagagcc ccaagaaaaa gagaaaggtg tgaccgcgga
     5941 tctggttacc actaaaccag cctcaagaac acccgaatgg agtctctaag ctacataata
     6001 ccaacttaca ctttacaaaa tgttgtcccc caaaatgtag ccattcgtat ctgctcctaa
     6061 taaaaagaaa gtttcttcac attctaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     6121 aaaaamcccc ccccccccct gcagggccaa gtcggcctct agaggtaccc aattcgccct
     6181 atagtgagtc gtattacaat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc
     6241 ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata
     6301 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc
     6361 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     6421 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
     6481 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     6541 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
     6601 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     6661 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     6721 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     6781 cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg
     6841 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ctgctcatga
     6901 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     6961 atttccgtgt cgcccttatt cccctttttg cggcattttg ccttcctgtt tttgctcacc
     7021 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     7081 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     7141 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     7201 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     7261 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     7321 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     7381 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     7441 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     7501 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     7561 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     7621 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     7681 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     7741 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     7801 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     7861 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     7921 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttc
//
