LOCUS       sequence-442603-        7907 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Adenine base editor with modified TadA variant for extended window 
            for A-to-G base editing in zebrafish.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7907)
  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 7907)
  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-ex1
FEATURES             Location/Qualifiers
     source          1..7907
                     /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             5723..5731
                     /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             5844..5864
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     3'UTR           5889..6014
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
     primer_bind     complement(6106..6125)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6107..6125)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6132..6149)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6132..6148)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6141..6163)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(6290..6745)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6427..6448)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6639..6658
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6772..6876
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6877..6945,6946..7737)
                     /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(7095..7114)
                     /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 catgagcgat aagaagtata
     1201 gcatcggcct ggctattgga actaactccg tgggttgggc agtgattaca gacgactaca
     1261 aggtccctag caagaaattt aaggtgctgg gtaacaccga caggcacagc atcaagaaaa
     1321 atctgattgg agccctgctg ttcggttctg gagagactgc cgaagcaaca cgcctgaaaa
     1381 gaacagcaag aaggcgctat accagaagga agaatagaat ctgttacctg caggagattt
     1441 tctctaacga aatggctaag gtggacgatt cattctttca taggctggag gaaagtttcc
     1501 tggtcgagga agataagaaa cacgagcgcc atcctatctt tggaaacatt gtggacgagg
     1561 tcgcctatca cgaaaaatac ccaaccatct atcatctgcg caagaaactg gctgactcta
     1621 ctgataaagc cgacctgaga ctgatctatc tggctctggc ccacatgatt aagttcaggg
     1681 gtcattttct gatcgagggc gatctgaacc ccgacaattc cgatgtggac aagctgttca
     1741 tccagctggt ccagatttac aatcagctgt ttgaggaaaa ccctattaat gcttccagag
     1801 tggacgcaaa agctatcctg tcagccaggc tgtccaagtc acgcagactg gagaacctga
     1861 ttgcacagct gcccggagaa aagaggaacg gtctgtttgg aaatctgatc gctctgagtc
     1921 tgggcctgac tcctaacttc aaaagcaatt ttgatctggc tgaggacgcc aaactgcagc
     1981 tgtcaaagga cacatatgac gatgacctgg ataacctgct ggcacagatc ggagatcagt
     2041 acgctgacct gttcctggct gccaaaaatc tgtccgacgc aatcctgctg tcagatattc
     2101 tgagagtgaa cagcgagatt acaaaagctc ctctgagtgc cagcatgatc aagagatatg
     2161 acgagcacca tcaggatctg accctgctga aggctctggt caggcagcag ctgccagaga
     2221 agtacaagga aattttcttt gatcagtcca agaacggcta cgccggttat atcgacggag
     2281 gcgcatcaca ggaggaattc tacaagttta tcaaacctat tctggagaag atggacggaa
     2341 ctgaggaact gctggtgaaa ctgaatagag aggacctgct gaggaagcag cgcacatttg
     2401 ataacggttc catcccacac cagattcatc tgggagagct gcacgctatc ctgaggcgcc
     2461 aggaagactt ctaccccttt ctgaaagata accgcgagaa gatcgaaaaa attctgacct
     2521 tcagaatccc ttactatgtg ggtccactgg ctcgcggaaa cagcagattt gcctggatga
     2581 ctcgcaaatc cgaggaaacc attactcctt ggaacttcga ggaagtggtc gataagggcg
     2641 cctctgcaca gtccttcatc gagagaatga ctaattttga caaaaacctg cccaatgaga
     2701 aagtgctgcc taagcactcc ctgctgtacg agtatttcac tgtctataac gaactgacaa
     2761 aggtgaaata cgtcaccgag ggcatgagaa agccagcctt cctgtcagga gagcagaaga
     2821 aagcaatcgt ggatctgctg tttaaaacca acaggaaagt gactgtcaag cagctgaagg
     2881 aggactactt caagaaaatt gaatgcttcg attccgtgga gatcagcgga gtcgaagaca
     2941 gatttaacgc aagcctgggc gcttaccacg atctgctgaa gatcattaag gataaagact
     3001 tcctggacaa cgaggaaaat gaggatatcc tggaagacat tgtgctgaca ctgaccctgt
     3061 ttgaggacag aggaatgatc gaggaaagac tgaaaaccta tgctcatctg ttcgatgaca
     3121 aggtgatgaa acagctgaag agaaggcgct acactggctg gggtagactg agcaggaagc
     3181 tgatcaacgg cattagggat aaacagtcag gaaagacaat cctggacttt ctgaaaagtg
     3241 atggcttcgc caaccgcaat tttatgcagc tgattcacga tgacagtctg accttcaaag
     3301 aggacatcca gaaggctcag gtgtctggac agggccactc cctgcatgag cagattgcaa
     3361 acctggctgg aagcccagcc atcaagaaag gcattctgca gacagtgaaa atcgtcgatg
     3421 agctggtgaa agtcatgggc cataagcccg aaaacatcgt gattgagatg gctcgcgaaa
     3481 atcagacaac ccagaagggt cagaagaaca gtagagagag gatgaaaaga atcgaggaag
     3541 gcattaagga gctgggtagc cagatcctga aagagcaccc agtggaaaac acacagctgc
     3601 agaatgagaa gctgtatctg tactatctgc agaatggaag agatatgtac gtggaccagg
     3661 agctggatat taacaggctg tctgattacg acgtggatca tatcgtcccc cagagtttca
     3721 tcaaagatga cagcattgac aacaaggtgc tgaccaggtc cgacaaaaac agaggaaaat
     3781 cagataatgt ccctagtgag gaagtggtca agaaaatgaa gaactactgg agacagctgc
     3841 tgaatgccaa actgatcact cagaggaagt ttgataacct gacaaaagca gagcgcggtg
     3901 gactgtcaga actggacaaa gctggattca tcaagaggca gctggtggaa acacgccaga
     3961 tcactaaaca cgtcgcacag attctggata gtcgcatgaa cacaaagtac gatgagaatg
     4021 acaaactgat cagagaagtg aaggtcatta ccctgaagag taaactggtc agcgacttta
     4081 ggaaagattt ccagttttat aaggtccgcg agattaacaa ttatcaccat gcccatgacg
     4141 catacctgaa cgccgtggtc ggtaccgcac tgatcaagaa atacccaaaa ctggagagcg
     4201 aattcgtgta cggagactat aaggtgtacg atgtcagaaa aatgatcgcc aagtccgagc
     4261 aggaaattgg aaaagctact gccaagtatt tcttttactc aaacatcatg aatttcttta
     4321 agacagagat caccctggcc aatggagaaa tccgcaaaag gcccctgatt gagacaaacg
     4381 gagagacagg cgaaatcgtg tgggacaaag gcagagattt tgcaaccgtg aggaaggtcc
     4441 tgagcatgcc tcaagtgaat atcgtcaaga aaactgaggt gcagacaggc ggtttctcaa
     4501 aagaaagtat tctgccaaaa cgcaactctg ataagctgat cgctagaaag aaagactggg
     4561 accctaagaa gtatggaggc tttgactctc ccactgtggc atactccgtc ctggtggtcg
     4621 ctaaggtgga gaagggcaaa agcaagaaac tgaaatctgt caaggagctg ctgggtatca
     4681 caattatgga gagaagctct ttcgagaaga acccaatcga ttttctggag gccaaaggtt
     4741 ataaggaagt gaagaaagac ctgatcatta aactgcccaa gtacagtctg tttgagctgg
     4801 aaaacggcag gaaacgcatg ctggcaagcg ctggagagct gcagaaaggc aatgaactgg
     4861 ccctgccttc taagtacgtg aacttcctgt atctggcaag ccactacgag aagctgaaag
     4921 gatctggaag ctccggttct agtagcgaag tcgagttttc acacgaatac tggatgagac
     4981 atgctctcac actcgcaaag agggctagag atgagaggga ggttcctgtg ggcgcagtgc
     5041 tggtcctgaa taacagggtt ataggcgagg gatggaaccg cgcaattgga ctgcatgacc
     5101 caaccgcaca cgcagagatt atggcactca ggcaaggagg actggtcatg cagaactata
     5161 gactcattga cgccacactg tatagcacct tcgagccttg tgtgatgtgc gcaggagcca
     5221 tgatccacag taggatcggt agagtcgtgt ttggagttag aaactcaaag cgcggagctg
     5281 ccggcagtct gatgaatgtt ctgaactacc caggaatgaa tcatagggtc gagatcaccg
     5341 aaggcattct ggccgacgaa tgtgccgctc tgctgtgcga tttctacaga atgccaagac
     5401 gggttttcaa cgctcagaag aaagcccagt cttccattaa tgcccgaacg ccagaggata
     5461 acgaacagaa acagctgttt gtggagcagc acaagcatta tctggacgag atcattgaac
     5521 agattagcga gttctctaaa agagtgatcc tggccgacgc aaatctggat aaggtcctgt
     5581 ctgcttacaa caaacacaga gataagccca tcagggagca ggccgaaaat atcattcatc
     5641 tgttcactct gacaaacctg ggcgcacctg cagctttcaa gtacttcgac actacaatcg
     5701 atagaaagag gtacacctcc actaaggagg tgctggacgc tacactgatc catcagagta
     5761 ttaccggcct gtacgaaaca aggattgacc tgtctcagct gggtggcgac aaaagaactg
     5821 cagatggatc cgagttcgag agccccaaga aaaagagaaa ggtgtgaccg cggatctggt
     5881 taccactaaa ccagcctcaa gaacacccga atggagtctc taagctacat aataccaact
     5941 tacactttac aaaatgttgt cccccaaaat gtagccattc gtatctgctc ctaataaaaa
     6001 gaaagtttct tcacawwmwa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaammcc
     6061 cccccccctg cagggccaag tcggcctcta gaggtaccca attcgcccta tagtgagtcg
     6121 tattacaatt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc
     6181 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc
     6241 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggca aattgtaagc
     6301 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     6361 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     6421 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     6481 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     6541 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     6601 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     6661 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     6721 cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     6781 cctatttgtt tatttttcta aatacattca aatatgtatc tgctcatgag acaataaccc
     6841 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     6901 gcccttattc ccctttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     6961 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     7021 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     7081 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     7141 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     7201 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     7261 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     7321 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     7381 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     7441 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     7501 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     7561 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     7621 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     7681 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     7741 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     7801 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     7861 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttc
//
