LOCUS       sequence-450069-        7550 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses the N-terminal of AID-N51A-CBE.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7550)
  AUTHORS   Liu Z, Chen S, Davis AE, Lo CH, Wang Q, Li T, Ning K, Zhang Q, Zhao 
            J, Wang S, Sun Y
  TITLE     Efficient Rescue of Retinal Degeneration in Pde6a Mice by Engineered
            Base Editing and Prime Editing.
  JOURNAL   Adv Sci (Weinh). 2024 Sep 19:e2405628. doi: 10.1002/advs.202405628.
  PUBMED    39297417
REFERENCE   2  (bases 1 to 7550)
  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: "Adv Sci 
            (Weinh). 2024 Sep 19:e2405628. doi: 10.1002/advs.202405628."
COMMENT     Sequence Label: AAV-AID-N51A-CBE N
FEATURES             Location/Qualifiers
     source          1..7550
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    5..21
                     /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     10..32
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     29..45
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     29..45
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   72..201
                     /label=AAV2 ITR
     enhancer        224..509
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     intron          789..1016
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     CDS             1730..3445
                     /codon_start=1
                     /product="N-terminal portion of Streptococcus pyogenes Cas9
                     (Zetsche et al., 2015)"
                     /label=Cas9(N)
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIE"
     CDS             3794..3814
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(3879..3899)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    4075..4299
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     misc_RNA        complement(4313..4388)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(4410..4658)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(4468..4487)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(4638..4658)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     repeat_region   4682..4811
                     /label=AAV2 ITR
     primer_bind     complement(4830..4847)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4830..4846)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4839..4861)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4988..5443
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5075..5094)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5285..5306
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5469..5573
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5574..5642,5643..6434)
                     /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     complement(5792..5811)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6605..7193
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7094..7113
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    7481..7502
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7517..7522,7523..7540,7541..7547)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
ORIGIN
        1 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaga
       61 tttaattaag gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
      121 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      181 actccatcac taggggttcc tgcggcctct agatcagggt acccgttaca taacttacgg
      241 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca atagtaacgc
      301 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
      361 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
      421 ggcccgcctg gcattgtgcc cagtacatga ccttatggga ctttcctact tggcagtaca
      481 tctacgtatt agtcatcgct attaccatgg tcgaggtgag ccccacgttc tgcttcactc
      541 tccccatctc ccccccctcc ccacccccaa ttttgtattt atttattttt taattatttt
      601 gtgcagcgat gggggcgggg gggggggggg ggcgcgcgcc gggggggggg gggggggggg
      661 gggggggggg sggggsgrgg sggagaggtg cggcggcagc caatcagagc ggcgcgctcc
      721 gaaagtttcc ttttatggcg aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc
      781 ggcgggcggg agtcgctgcg cgctgccttc gccccgtgcc ccgctccgcc gccgcctcgc
      841 gccgcccgcc ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc
      901 cttctcctcc gggctgtaat tagctgagca agaggtaagg gtttaaggga tggttggttg
      961 gtggggtatt aatgtttaat tacctggagc acctgcctga aatcactttt tttcaggttg
     1021 gaccggtgcc gccaccatga aacggacagc cgacggaagc gagttcgagt cagcaaagaa
     1081 gaagcggaaa gtcgatagcc tgctgatgaa ccgtcgtgaa tttctgtatc agtttaaaaa
     1141 cgtgcgttgg gcgaaaggcc gtcgtgaaac ctatctgtgc tatgtggtga aacgtcgtga
     1201 tagcgcgacc agctttagcc tggattttgg ctatctgcgt gccaaaaacg gctgccatgt
     1261 ggaactgctg tttctgcgtt atattagcga ttgggatctg gatccgggcc gttgctatcg
     1321 tgtgacctgg tttatcagct ggagcccgtg ctatgattgc gcgcgtcatg tggcggattt
     1381 tctgcgtggc aacccgaacc tgagcctgcg tatttttacc gcgcgtctgt atttttgcga
     1441 agccggcagg cgtgaaccgg aaggcctgcg tcgtctgcat cgtgcgggcg tgcagattgc
     1501 gattatgacc tttaaagatt atttttattg ctggaacacc tttgtggaaa accatggacg
     1561 tacctttaaa gcgtgggaag gcctgcatga aaacagcgtg cgtctgagcc gtcagctgcg
     1621 tcgtattctg ctgtctggcg gatctagcgg aggatcctct ggcagcgaga caccaggaac
     1681 aagcgagtca gcaacaccag agagcagtgg cggcagcagc ggcggcagcg acaagaagta
     1741 cagcatcggc ctggccatcg gcaccaactc tgtgggctgg gccgtgatca ccgacgagta
     1801 caaggtgccc agcaagaaat tcaaggtgct gggcaacacc gaccggcaca gcatcaagaa
     1861 gaacctgatc ggagccctgc tgttcgacag cggcgaaaca gccgaggcca cccggctgaa
     1921 gagaaccgcc agaagaagat acaccagacg gaagaaccgg atctgctatc tgcaagagat
     1981 cttcagcaac gagatggcca aggtggacga cagcttcttc cacagactgg aagagtcctt
     2041 cctggtggaa gaggataaga agcacgagcg gcaccccatc ttcggcaaca tcgtggacga
     2101 ggtggcctac cacgagaagt accccaccat ctaccacctg agaaagaaac tggtggacag
     2161 caccgacaag gccgacctgc ggctgatcta tctggccctg gcccacatga tcaagttccg
     2221 gggccacttc ctgatcgagg gcgacctgaa ccccgacaac agcgacgtgg acaagctgtt
     2281 catccagctg gtgcagacct acaaccagct gttcgaggaa aaccccatca acgccagcgg
     2341 cgtggacgcc aaggccatcc tgtctgccag actgagcaag agcagacggc tggaaaatct
     2401 gatcgcccag ctgcccggcg agaagaagaa tggcctgttc ggaaacctga ttgccctgag
     2461 cctgggcctg acccccaact tcaagagcaa cttcgacctg gccgaggatg ccaaactgca
     2521 gctgagcaag gacacctacg acgacgacct ggacaacctg ctggcccaga tcggcgacca
     2581 gtacgccgac ctgtttctgg ccgccaagaa cctgtccgac gccatcctgc tgagcgacat
     2641 cctgagagtg aacaccgaga tcaccaaggc ccccctgagc gcctctatga tcaagagata
     2701 cgacgagcac caccaggacc tgaccctgct gaaagctctc gtgcggcagc agctgcctga
     2761 gaagtacaaa gagattttct tcgaccagag caagaacggc tacgccggct acattgacgg
     2821 cggagccagc caggaagagt tctacaagtt catcaagccc atcctggaaa agatggacgg
     2881 caccgaggaa ctgctcgtga agctgaacag agaggacctg ctgcggaagc agcggacctt
     2941 cgacaacggc agcatccccc accagatcca cctgggagag ctgcacgcca ttctgcggcg
     3001 gcaggaagat ttttacccat tcctgaagga caaccgggaa aagatcgaga agatcctgac
     3061 cttccgcatc ccctactacg tgggccctct ggccagggga aacagcagat tcgcctggat
     3121 gaccagaaag agcgaggaaa ccatcacccc ctggaacttc gaggaagtgg tggacaaggg
     3181 cgcttccgcc cagagcttca tcgagcggat gaccaacttc gataagaacc tgcccaacga
     3241 gaaggtgctg cccaagcaca gcctgctgta cgagtacttc accgtgtata acgagctgac
     3301 caaagtgaaa tacgtgaccg agggaatgag aaagcccgcc ttcctgagcg gcgagcagaa
     3361 aaaggccatc gtggacctgc tgttcaagac caaccggaaa gtgaccgtga agcagctgaa
     3421 agaggactac ttcaagaaaa tcgagtgcct gtcctacgag acagagatcc tgacagtgga
     3481 gtatggcctg ctgccaatcg gcaagatcgt ggagaagagg atcgagtgta ccgtgtactc
     3541 tgtggataac aatggcaaca tctatacaca gcccgtggca cagtggcacg ataggggaga
     3601 gcaggaggtg ttcgagtatt gcctggagga cggcagcctg atcagggcaa ccaaggacca
     3661 caagttcatg acagtggatg gccagatgct gcccatcgac gagattttcg agcgggagct
     3721 ggacctgatg agagtggata acctgcctaa tagcggaggc agtaaaagaa cagcagacgg
     3781 gagtgagttt gagcccaaga aaaagagaaa ggtgtaagat ctgataatca acctctggat
     3841 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     3901 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     3961 tcctccttgt ataaatcctg gttagttctt gccacggcgg aactcatcgc cgcctgcctt
     4021 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gcgactgtgc
     4081 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     4141 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     4201 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
     4261 acaatagcag gcatgctggg gatgcggtgg gctctatggc tcgagaaaaa aagcaccgac
     4321 tcggtgccac tttttcaagt tgataacgga ctagccttat tttaacttgc tatttctagc
     4381 tctaaaaccg agacgcatta atcgtctcgg gtgtttcgtc ctttccacaa gatatataaa
     4441 gccaagaaat cgaaatactt tcaagttacg gtaagcatat gatagtccat tttaaaacat
     4501 aattttaaaa ctgcaaacta cccaagaaat tattactttc tacgtcacgt attttgtact
     4561 aatatctttg tgtttacagt caaattaatt ctaattatct ctctaacagc cttgtatcgt
     4621 atatgcaaat atgaaggaat catgggaaat aggccctctt cctgcccgac cttgcggccg
     4681 caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag
     4741 gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag
     4801 cgagcgcgca gccttaatta acctaattca ctggccgtcg ttttacaacg tcgtgactgg
     4861 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     4921 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     4981 gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     5041 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     5101 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     5161 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     5221 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     5281 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     5341 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     5401 aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg
     5461 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     5521 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     5581 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     5641 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     5701 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     5761 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     5821 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     5881 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     5941 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     6001 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     6061 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     6121 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     6181 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     6241 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     6301 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     6361 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     6421 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     6481 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     6541 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     6601 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     6661 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     6721 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     6781 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     6841 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     6901 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     6961 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     7021 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     7081 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     7141 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     7201 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     7261 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     7321 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     7381 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
     7441 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
     7501 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
//
