LOCUS       sequence-394768-        7392 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of N-terminally fused Nme2-ABE8e-nt.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7392)
  AUTHORS   Bamidele N, Zhang H, Dong X, Cheng H, Gaston N, Feinzig H, Cao H, 
            Kelly K, Watts JK, Xie J, Gao G, Sontheimer EJ
  TITLE     Domain-inlaid Nme2Cas9 adenine base editors with improved activity 
            and targeting scope.
  JOURNAL   Nat Commun. 2024 Feb 17;15(1):1458. doi: 10.1038/s41467-024-45763-5.
  PUBMED    38368418
REFERENCE   2  (bases 1 to 7392)
  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: "2024-02-17"; volume: "15(1):1458. doi"; pages: " 
            10.1038/s41467-024-45763-5"
COMMENT     Sequence Label: pEJS2156_Nme2-ABE8e-nt
FEATURES             Location/Qualifiers
     source          1..7392
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     610..629
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        610..628
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             678..698
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4611..4631
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4640..4657
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4680..4697)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4686..4910
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4981..4997)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4981..4997)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4994..5016)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5005..5021
                     /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(5029..5035,5036..5053,5054..5059))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5074..5095
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5212..5229)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5383..5971)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5463..5482)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6142..6933,6934..7002))
                     /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     6765..6784
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7003..7107)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7186..7205)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        join(7377..7392,1..364)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagagatc
      601 cgcggccgct aatacgactc actataggga gagccgccac catgaaacgg acagccgacg
      661 gaagcgagtt cgagtcacca aagaagaagc ggaaagtcgg cggtagcggc ggaggcagcg
      721 gtggcggcag cggctctgag gtggagtttt cccacgagta ctggatgaga catgccctga
      781 ccctggccaa gagggcacgc gatgagaggg aggtgcctgt gggagccgtg ctggtgctga
      841 acaatagagt gatcggcgag ggctggaaca gagccatcgg cctgcacgac ccaacagccc
      901 atgccgaaat tatggccctg agacagggcg gcctggtcat gcagaactac agactgattg
      961 acgccaccct gtacgtgaca ttcgagcctt gcgtgatgtg cgccggcgcc atgatccact
     1021 ctaggatcgg ccgcgtggtg tttggcgtga ggaacagcaa acggggcgcc gcaggctccc
     1081 tgatgaacgt gctgaactac cccggcatga atcaccgcgt cgaaattacc gagggaatcc
     1141 tggcagatga atgtgccgcc ctgctgtgcg acttctaccg gatgcctaga caggtgttca
     1201 atgctcagaa gaaggcccag agctccatca actccggagg atctagcgga ggctcctctg
     1261 gctctgagac acctggcaca agcgagagcg caacacctga aagcagcggg ggcagcagcg
     1321 gggggtcaat ggccgccttc aagcctaacc caatcaatta catcctggga ctggccatcg
     1381 gaatcgcatc cgtgggatgg gctatggtgg agatcgacga ggaggagaat cctatccggc
     1441 tgatcgatct gggcgtgaga gtgtttgaga gggccgaggt gccaaagacc ggcgattctc
     1501 tggctatggc ccggagactg gcacggagcg tgaggcgcct gacacggaga agggcacaca
     1561 ggctgctgag ggcacgccgg ctgctgaaga gagagggcgt gctgcaggca gcagacttcg
     1621 atgagaatgg cctgatcaag agcctgccaa acaccccctg gcagctgaga gcagccgccc
     1681 tggacaggaa gctgacacca ctggagtggt ctgccgtgct gctgcacctg atcaagcacc
     1741 gcggctacct gagccagcgg aagaacgagg gagagacagc agacaaggag ctgggcgccc
     1801 tgctgaaggg agtggccaac aatgcccacg ccctgcagac cggcgatttc aggacacctg
     1861 ccgagctggc cctgaataag tttgagaagg agtccggcca catcagaaac cagaggggcg
     1921 actatagcca caccttctcc cgcaaggatc tgcaggccga gctgatcctg ctgttcgaga
     1981 agcagaagga gtttggcaat ccacacgtga gcggaggcct gaaggaggga atcgagaccc
     2041 tgctgatgac acagaggcct gccctgtccg gcgacgcagt gcagaagatg ctgggacact
     2101 gcaccttcga gcctgcagag ccaaaggccg ccaagaacac ctacacagcc gagcggttta
     2161 tctggctgac aaagctgaac aatctgagaa tcctggagca gggatccgag aggccactga
     2221 ccgacacaga gagggccacc ctgatggatg agccttaccg gaagtctaag ctgacatatg
     2281 cccaggccag aaagctgctg ggcctggagg acaccgcctt ctttaagggc ctgagatacg
     2341 gcaaggataa tgccgaggcc tccacactga tggagatgaa ggcctatcac gccatctctc
     2401 gcgccctgga gaaggagggc ctgaaggaca agaagtcccc cctgaacctg agctccgagc
     2461 tgcaggatga gatcggcacc gccttctctc tgtttaagac cgacgaggat atcacaggcc
     2521 gcctgaagga cagggtgcag cctgagatcc tggaggccct gctgaagcac atctctttcg
     2581 ataagtttgt gcagatcagc ctgaaggccc tgagaaggat cgtgccactg atggagcagg
     2641 gcaagcggta cgacgaggcc tgcgccgaga tctacggcga tcactatggc aagaagaaca
     2701 cagaggagaa gatctatctg ccccctatcc ctgccgacga gatcagaaat cctgtggtgc
     2761 tgagggccct gtcccaggca agaaaagtga tcaacggagt ggtgcgccgg tacggatctc
     2821 cagcccggat ccacatcgag accgccagag aagtgggcaa gagcttcaag gaccggaagg
     2881 agatcgagaa gagacaggag gagaatcgca aggatcggga gaaggccgcc gccaagttta
     2941 gggagtactt ccctaacttt gtgggcgagc caaagtctaa ggacatcctg aagctgcgcc
     3001 tgtacgagca gcagcacggc aagtgtctgt atagcggcaa ggagatcaat ctggtgcggc
     3061 tgaacgagaa gggctatgtg gagatcgatc acgccctgcc tttctccaga acctgggacg
     3121 attcttttaa caataaggtg ctggtgctgg gcagcgagaa ccagaataag ggcaatcaga
     3181 caccatacga gtatttcaat ggcaaggaca actccaggga gtggcaggag ttcaaggccc
     3241 gcgtggagac ctctagattt cccaggagca agaagcagcg gatcctgctg cagaagttcg
     3301 acgaggatgg ctttaaggag tgcaacctga atgacaccag atacgtgaac cggttcctgt
     3361 gccagtttgt ggccgatcac atcctgctga ccggcaaggg caagagaagg gtgttcgcct
     3421 ctaatggcca gatcacaaac ctgctgaggg gattttgggg actgaggaag gtgcgggcag
     3481 agaatgacag acaccacgca ctggatgcag tggtggtggc atgcagcacc gtggcaatgc
     3541 agcagaagat cacaagattc gtgaggtata aggagatgaa cgcctttgac ggcaagacca
     3601 tcgataagga gacaggcaag gtgctgcacc agaagaccca cttcccccag ccttgggagt
     3661 tctttgccca ggaagtgatg atccgggtgt tcggcaagcc agacggcaag cctgagtttg
     3721 aggaggccga taccccagag aagctgagga cactgctggc agagaagctg tctagcaggc
     3781 cagaggcagt gcacgagtac gtgaccccac tgttcgtgtc cagggcaccc aatcggaaga
     3841 tgtctggcgc ccacaaggac acactgagaa gcgccaagag gtttgtgaag cacaacgaga
     3901 agatctccgt gaagagagtg tggctgaccg agatcaagct ggccgatctg gagaacatgg
     3961 tgaattacaa gaacggcagg gagatcgagc tgtatgaggc cctgaaggca aggctggagg
     4021 cctacggagg aaatgccaag caggccttcg acccaaagga taaccccttt tataagaagg
     4081 gaggacagct ggtgaaggcc gtgcgggtgg agaagaccca ggagagcggc gtgctgctga
     4141 ataagaagaa cgcctacaca atcgccgaca atggcgatat ggtgagagtg gacgtgttct
     4201 gtaaggtgga taagaagggc aagaatcagt actttatcgt gcctatctat gcctggcagg
     4261 tggccgagaa catcctgcca gacatcgatt gcaagggcta cagaatcgac gatagctata
     4321 cattctgttt ttccctgcac aagtatgacc tgatcgcctt ccagaaggat gagaagtcca
     4381 aggtggagtt tgcctactat atcaattgcg actcctctaa cggcaggttc tacctggcct
     4441 ggcacgataa gggcagcaag gagcagcagt ttcgcatctc cacccagaat ctggtgctga
     4501 tccagaagta tcaggtgaac gagctgggca aggagatcag gccatgtcgg ctgaagaagc
     4561 gcccacccgt gcgggaggat aaaagaaccg ccgacggcag cgaattcgag cccaagaaga
     4621 agaggaaagt ctaaccggtc atcatcacca tcaccattga gtttaaaccc gctgatcagc
     4681 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     4741 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     4801 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     4861 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc
     4921 ggaaagaacc agctggggct cgataccgtc gacctctagc tagagcttgg cgtaatcatg
     4981 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     5041 cggaagcata aagtgtaaag cctagggtgc ctaatgagtg agctaactca cattaattgc
     5101 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     5161 cggccaacgc gcggggagag gcggtttgcg tattgggcgc acttccgctt cctcgctcac
     5221 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     5281 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     5341 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     5401 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     5461 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     5521 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     5581 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     5641 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     5701 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     5761 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     5821 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     5881 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     5941 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     6001 tgacactcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     6061 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     6121 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     6181 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     6241 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgggacccac gctcaccggc
     6301 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     6361 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     6421 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     6481 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     6541 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     6601 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     6661 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     6721 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     6781 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     6841 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     6901 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     6961 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     7021 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     7081 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga
     7141 cggatcggga gatcgatctc ccgatcccct agggtcgact ctcagtacaa tctgctctga
     7201 tgccgcatag ttaaaccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
     7261 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
     7321 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
     7381 ttgattattg ac
//
