LOCUS       sequence-375358-        5780 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of nuclear v8 ZF-DdCBE in mammalian cells (C-terminal 
            DddAC, Left).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5780)
  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 5780)
  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: pL7_3xG35-EMILIN2_X1
FEATURES             Location/Qualifiers
     source          1..5780
                     /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      2012..2440
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2099..2118)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2309..2330
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2449..2469)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2454..2783
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2634..2769
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2696..2715
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        2831..2878
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2897..3295
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    3453..3574
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3490..3509)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3544..3563
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3623..3639)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3623..3639)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3636..3658)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3647..3663
                     /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(3671..3677,3678..3695,3696..3701))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3716..3737
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3854..3871)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4025..4613)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4105..4124)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4784..5575,5576..5644))
                     /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     5407..5426
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5645..5749)
                     /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 cccaggggct ttggttcggc acatccgcac ccacactggc
     1201 gaaaagccgt ttgcgtgtga catctgcgga cgcaagtttg cgcgctctga taacctggtc
     1261 agacatataa ggacacacac cggtgagaaa cccttcgctt gcgacatttg cgggagaaag
     1321 tttgcaacaa gcggctctct cgttagacat 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 ataaaccacg atcgttatgc tgatcatacc ctaatgatcc
     1801 cagcaagata atgtcctgtc ttctaagatg tgcatcaagc ctggtacata ctgaaaaccc
     1861 tataaggtcc tggataattt ttgtttgatt attcattgaa gaaacattta ttttccaatt
     1921 gtgtgaagtt tttgactgtt aataaaagaa tctgtcaacc atcaaacttc tgaggcggaa
     1981 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
     2041 gcgggtgtgg tggtcacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     2101 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     2161 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     2221 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     2281 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     2341 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     2401 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
     2461 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     2521 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     2581 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     2641 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     2701 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     2761 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg
     2821 atcagcacgt gttgacaatt aatcatcggc atagtatatc ggcatagtat aatacggcaa
     2881 ggtgaggaac taaaccatgg ccaagccttt gtctcaagaa gaatccaccc tcattgaaag
     2941 agcaacggct acaatcaaca gcatccccat ctctgaagac tacagcgtcg ccagcgcagc
     3001 tctctctagc gacggccgca tcttcactgg tgtcaatgta tatcatttta ctgggggacc
     3061 ttgtgcagaa ctcgtggtgc tgggcactgc tgctgctgcg gcagctggca acctgacttg
     3121 tatcgtcgcg atcggaaatg agaacagggg catcttgagc ccctgcggac ggtgccgaca
     3181 ggtgcttctc gatctgcatc ctgggatcaa agccatagtg aaggacagtg atggacagcc
     3241 gacggcagtt gggattcgtg aattgctgcc ctctggttat gtgtgggagg gctaagcact
     3301 tcgtggccga ggagcaggac tgacacgtgc tacgagattt cgattccacc gccgccttct
     3361 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     3421 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     3481 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     3541 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     3601 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     3661 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     3721 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     3781 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     3841 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     3901 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     3961 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     4021 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     4081 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     4141 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     4201 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     4261 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     4321 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     4381 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     4441 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     4501 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     4561 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     4621 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     4681 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     4741 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     4801 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     4861 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     4921 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     4981 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     5041 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     5101 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     5161 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     5221 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     5281 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     5341 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     5401 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     5461 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     5521 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     5581 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     5641 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     5701 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     5761 gaaaagtgcc acctgacgtc
//
