LOCUS       sequence-375374-        6014 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of nuclear v8 ZF-DdCBE in mammalian cells (C-terminal 
            DddAN, Left).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6014)
  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 6014)
  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: pL5_3xG22-DCAF8L2_X1
FEATURES             Location/Qualifiers
     source          1..6014
                     /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             1744..1995
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     rep_origin      2246..2674
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2333..2352)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2543..2564
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2683..2703)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2688..3017
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2868..3003
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2930..2949
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3065..3112
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3131..3529
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    3687..3808
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3724..3743)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3778..3797
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3857..3873)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3857..3873)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3870..3892)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3881..3897
                     /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(3905..3911,3912..3929,3930..3935))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3950..3971
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4088..4105)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4259..4847)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4339..4358)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5018..5809,5810..5878))
                     /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     5641..5660
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5879..5983)
                     /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 aattcgctcg ctctgataac ctggtcagac acatccgcac ccacactggc
     1201 gaaaagccgt ttgcgtgtga catctgcgga cgcaagtttg cgcgggaaga caaccttcat
     1261 acgcatataa ggacacacac cggtgagaaa cccttcgctt gcgacatttg cgggagaaag
     1321 tttgcacgga gcgacaacct cgtcaggcat accaagatac acctgcgggg atccggcgga
     1381 ggaggttctg gcggttctgg aggaagtggc agctacgccc tgggtccgta tcagattagc
     1441 gccccgcagc tgccagctta caatggtcag accgtgggta ccttctacta tgtgaacgac
     1501 gcgggcggtc tggagagcaa ggtgtttagc agcggcggtc caaccccgta cccaaactat
     1561 gccaatgccg gtcatgtgga gggtcagagc gccctgttca tgcgtgataa cggcatcagc
     1621 gagggtctgg tgttccacaa caacccggaa ggcacctgcg gtttttgcgt gaacatgatc
     1681 gagaccctgc tgccggaaaa cgcgaaaatg accgtggtgc cgccgaaagg ttctggcggc
     1741 tcaactaatc tgagcgacat cattgagaag gagactggga aacagctggt cattcaggag
     1801 tccatcctga tgctgcctga ggaggtggag gaagtgatcg gcaacaagcc agagtctgac
     1861 atcctggtgc acaccgccta cgacgagtcc acagatgaga atgtgatgct gctgacctct
     1921 gacgcccccg agtataagcc ttgggccctg gtcatccagg attctaacgg cgagaataag
     1981 atcaagatgc tgtgataaac cacgatcgtt atgctgatca taccctaatg atcccagcaa
     2041 gataatgtcc tgtcttctaa gatgtgcatc aagcctggta catactgaaa accctataag
     2101 gtcctggata atttttgttt gattattcat tgaagaaaca tttattttcc aattgtgtga
     2161 agtttttgac tgttaataaa agaatctgtc aaccatcaaa cttctgaggc ggaaagaacc
     2221 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     2281 gtggtggtca cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     2341 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     2401 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     2461 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     2521 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     2581 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     2641 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
     2701 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     2761 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     2821 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     2881 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     2941 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     3001 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcagc
     3061 acgtgttgac aattaatcat cggcatagta tatcggcata gtataatacg gcaaggtgag
     3121 gaactaaacc atggccaagc ctttgtctca agaagaatcc accctcattg aaagagcaac
     3181 ggctacaatc aacagcatcc ccatctctga agactacagc gtcgccagcg cagctctctc
     3241 tagcgacggc cgcatcttca ctggtgtcaa tgtatatcat tttactgggg gaccttgtgc
     3301 agaactcgtg gtgctgggca ctgctgctgc tgcggcagct ggcaacctga cttgtatcgt
     3361 cgcgatcgga aatgagaaca ggggcatctt gagcccctgc ggacggtgcc gacaggtgct
     3421 tctcgatctg catcctggga tcaaagccat agtgaaggac agtgatggac agccgacggc
     3481 agttgggatt cgtgaattgc tgccctctgg ttatgtgtgg gagggctaag cacttcgtgg
     3541 ccgaggagca ggactgacac gtgctacgag atttcgattc caccgccgcc ttctatgaaa
     3601 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     3661 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
     3721 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     3781 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
     3841 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     3901 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     3961 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     4021 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     4081 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     4141 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     4201 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     4261 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     4321 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     4381 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     4441 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     4501 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     4561 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     4621 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     4681 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     4741 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     4801 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     4861 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     4921 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     4981 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     5041 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     5101 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     5161 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     5221 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     5281 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     5341 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     5401 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     5461 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     5521 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     5581 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     5641 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     5701 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     5761 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     5821 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     5881 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     5941 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     6001 tgccacctga cgtc
//
