LOCUS       sequence-389974-        6007 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of nuclear ZF-DdCBE in mammalian cells (N-terminal DddAC,
            Right).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6007)
  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 6007)
  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: pL8N_RB610-HBB_X1
FEATURES             Location/Qualifiers
     source          1..6007
                     /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             1792..2043
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     rep_origin      2240..2668
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2327..2346)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2537..2558
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2677..2697)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2682..3011
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2862..2997
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2924..2943
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3059..3106
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3125..3523
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    3681..3802
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3718..3737)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3772..3791
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3851..3867)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3851..3867)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3864..3886)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3875..3891
                     /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(3899..3905,3906..3923,3924..3929))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3944..3965
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4082..4099)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4252..4840)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4332..4351)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5011..5802,5803..5871))
                     /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     5634..5653
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5872..5976)
                     /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 tctggcggct cagccattcc agtgaagcgc
     1141 ggcgctaccg gtgaaaccaa agtgtttatc ggtaacagca acagcccgaa gagcccgacc
     1201 aaaggcggtt gcggatccgg cggaggaggt tctggcggtt ctggaggaag tatggctgag
     1261 aggcccttcg cctgcgatat atgtggcagg aaattcgcta gactcaggga tatacaattt
     1321 cacatccgca cccacactgg cgaaaagccg tttgcgtgtg acatctgcgg acgcaagttt
     1381 gcggatcccg gacacttggt tcgacatata aggacacaca ccggtgagaa acccttcgct
     1441 tgcgacattt gcgggagaaa gtttgcaact agcgggaacc tcgtccggca cattagaact
     1501 catacagggg aaaaaccatt tgcatgtgat atctgtggtc ggaaattcgc ccagaaatct
     1561 tctttgatag ctcatatccg aacgcatacg ggagagaagc cttttgcttg tgacatatgt
     1621 ggacgaaaat tcgcacaaag cggggacttg agaaggcata ttcgcaccca tacaggagaa
     1681 aagcccttcg catgcgatat ttgcggtcgt aagtttgctc aagcctctaa cttgattagc
     1741 cataccaaga tacacctgcg gagcggagga tccggaggat ctggaggcag cactaatctg
     1801 agcgacatca ttgagaagga gactgggaaa cagctggtca ttcaggagtc catcctgatg
     1861 ctgcctgagg aggtggagga agtgatcggc aacaagccag agtctgacat cctggtgcac
     1921 accgcctacg acgagtccac agatgagaat gtgatgctgc tgacctctga cgcccccgag
     1981 tataagcctt gggccctggt catccaggat tctaacggcg agaataagat caagatgctg
     2041 tgataagggg tctttgtcct ctgtactgtc tctctccttg cccctaaccc aaaaagcttc
     2101 atttttctgt gtaggctgca caagagcctt gattgaagat atattctttc tgaacagtat
     2161 ttaaggtttc caataaaatg tacacccctc agaacttctg aggcggaaag aaccagctgg
     2221 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     2281 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     2341 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     2401 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     2461 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     2521 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     2581 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     2641 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg
     2701 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     2761 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     2821 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     2881 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     2941 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     3001 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat cagcacgtgt
     3061 tgacaattaa tcatcggcat agtatatcgg catagtataa tacgacaagg tgaggaacta
     3121 aaccatggcc aagcctttgt ctcaagaaga atccaccctc attgaaagag caacggctac
     3181 aatcaacagc atccccatct ctgaagacta cagcgtcgcc agcgcagctc tctctagcga
     3241 cggccgcatc ttcactggtg tcaatgtata tcattttact gggggacctt gtgcagaact
     3301 cgtggtgctg ggcactgctg ctgctgcggc agctggcaac ctgacttgta tcgtcgcgat
     3361 cggaaatgag aacaggggca tcttgagccc ctgcggacgg tgccgacagg tgcttctcga
     3421 tctgcatcct gggatcaaag ccatagtgaa ggacagtgat ggacagccga cggcagttgg
     3481 gattcgtgaa ttgctgccct ctggttatgt gtgggagggc taagcacttc gtggccgagg
     3541 agcaggactg acacgtgcta cgagatttcg attccaccgc cgccttctat gaaaggttgg
     3601 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     3661 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca
     3721 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     3781 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg
     3841 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     3901 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     3961 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     4021 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     4081 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     4141 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     4201 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggct ttttccatag
     4261 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     4321 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     4381 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     4441 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     4501 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     4561 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     4621 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     4681 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     4741 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     4801 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     4861 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     4921 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     4981 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     5041 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     5101 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     5161 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     5221 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     5281 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     5341 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     5401 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     5461 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     5521 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     5581 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     5641 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     5701 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     5761 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     5821 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     5881 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     5941 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     6001 tgacgtc
//
