LOCUS       sequence-375357-        5724 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of nuclear v8 ZF-DdCBE in mammalian cells (C-terminal 
            DddAC, Right).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5724)
  AUTHORS   Willis JCW, Silva-Pinheiro P, Widdup L, Minczuk M, Liu DR
  TITLE     Compact zinc finger base editors that edit mitochondrial or nuclear 
            DNA in vitro and in vivo.
  JOURNAL   Nat Commun. 2022 Nov 23;13(1):7204. doi: 10.1038/s41467-022-34784-7.
  PUBMED    36418298
REFERENCE   2  (bases 1 to 5724)
  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: "2022-11-23"; volume: "13(1):7204. doi"; pages: " 
            10.1038/s41467-022-34784-7"
COMMENT     Sequence Label: pL8_3xG34-DCAF8L2_X1
FEATURES             Location/Qualifiers
     source          1..5724
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             940..960
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             970..996
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             1039..1059
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1069..1095
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             1510..1761
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     rep_origin      1958..2386
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2045..2064)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2255..2276
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2395..2415)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2400..2729
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2580..2715
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2642..2661
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        2777..2824
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     polyA_signal    3397..3518
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3434..3453)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3488..3507
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3567..3583)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3567..3583)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3580..3602)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3591..3607
                     /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(3615..3621,3622..3639,3640..3645))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3660..3681
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3798..3815)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3969..4557)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4049..4068)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4728..5519,5520..5588))
                     /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     5351..5370
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5589..5693)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 accatgaaac ggacagccga cggaagcgag ttcgagtcac caaagaagaa gcggaaagtc
      961 agcggatctc ctgcagctaa gagagtgaag ctggactctg gttcaaagag aactgctgat
     1021 ggttctgaat ttgaatctcc taaaaaaaaa agaaaggttt caggcagccc agctgcaaaa
     1081 agggttaaat tggatagtgg gtccgccgct atggctgaga ggcccttcgc ctgcgatata
     1141 tgtggcagga aattcgctga ttgccgagat cttgctcggc acatccgcac ccacactggc
     1201 gaaaagccgt ttgcgtgtga catctgcgga cgcaagtttg cgcgctctga taacctggtc
     1261 agacatataa ggacacacac cggtgagaaa cccttcgctt gcgacatttg cgggagaaag
     1321 tttgcacggt cagatgagct ggtacgccat accaagatac acctgcgggg atccggcgga
     1381 ggaggttctg gcggttctgg aggaagtgcc attccagtga agcgcggcgc taccggtgaa
     1441 accaaagtgt ttatcggtaa cagcaacagc ccgaagagcc cgaccaaagg cggttgctct
     1501 ggcggctcaa ctaatctgag cgacatcatt gagaaggaga ctgggaaaca gctggtcatt
     1561 caggagtcca tcctgatgct gcctgaggag gtggaggaag tgatcggcaa caagccagag
     1621 tctgacatcc tggtgcacac cgcctacgac gagtccacag atgagaatgt gatgctgctg
     1681 acctctgacg cccccgagta taagccttgg gccctggtca tccaggattc taacggcgag
     1741 aataagatca agatgctgtg ataaggggtc tttgtcctct gtactgtctc tctccttgcc
     1801 cctaacccaa aaagcttcat ttttctgtgt aggctgcaca agagccttga ttgaagatat
     1861 attctttctg aacagtattt aaggtttcca ataaaatgta cacccctcag aacttctgag
     1921 gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta
     1981 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     2041 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     2101 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     2161 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     2221 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     2281 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     2341 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
     2401 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     2461 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     2521 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     2581 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     2641 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
     2701 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg
     2761 gatctgatca gcacgtgttg acaattaatc atcggcatag tatatcggca tagtataata
     2821 cgacaaggtg aggaactaaa ccatggccaa gcctttgtct caagaagaat ccaccctcat
     2881 tgaaagagca acggctacaa tcaacagcat ccccatctct gaagactaca gcgtcgccag
     2941 cgcagctctc tctagcgacg gccgcatctt cactggtgtc aatgtatatc attttactgg
     3001 gggaccttgt gcagaactcg tggtgctggg cactgctgct gctgcggcag ctggcaacct
     3061 gacttgtatc gtcgcgatcg gaaatgagaa caggggcatc ttgagcccct gcggacggtg
     3121 ccgacaggtg cttctcgatc tgcatcctgg gatcaaagcc atagtgaagg acagtgatgg
     3181 acagccgacg gcagttggga ttcgtgaatt gctgccctcg gtatgtgtgg gagggctaag
     3241 cacttcgtgg ccgaggagca ggactgacac gtgctacgag atttcgattc caccgccgcc
     3301 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     3361 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat
     3421 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     3481 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc
     3541 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     3601 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     3661 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     3721 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     3781 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     3841 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     3901 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     3961 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     4021 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     4081 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     4141 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     4201 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     4261 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     4321 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     4381 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     4441 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     4501 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     4561 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     4621 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     4681 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     4741 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     4801 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     4861 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     4921 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     4981 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     5041 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     5101 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     5161 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     5221 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     5281 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     5341 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     5401 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     5461 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     5521 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     5581 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     5641 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     5701 ccccgaaaag tgccacctga cgtc
//
