LOCUS       sequence-410676-        7095 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for bacterial purification of codon optimized ABE8e-Cas9NG.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7095)
  AUTHORS   Martin-Rufino JD, Castano N, Pang M, Grody EI, Joubran S, Caulier A,
            Wahlster L, Li T, Qiu X, Riera-Escandell AM, Newby GA, Al'Khafaji A,
            Chaudhary S, Black S, Weng C, Munson G, Liu DR, Wlodarski MW, Sims 
            K, Oakley JH, Fasano RM, Xavier RJ, Lander ES, Klein DE, Sankaran VG
  TITLE     Massively parallel base editing to map variant effects in human 
            hematopoiesis.
  JOURNAL   Cell. 2023 Apr 27:S0092-8674(23)00332-X. doi: 
            10.1016/j.cell.2023.03.035.
  PUBMED    37137305
REFERENCE   2  (bases 1 to 7095)
  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: "Cell. 2023 
            Apr 27:S0092-8674(23)00332-X. doi: 10.1016/j.cell.2023.03.035."
COMMENT     Sequence Label: ABE8e-Cas9NG
FEATURES             Location/Qualifiers
     source          1..7095
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..546
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     447..466
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     663..680
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(811..838)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(862..1671)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     1585..1604
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(1672..1762)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(1771..1801)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        1866..1984
                     /label=rhaB promoter
                     /note="promoter of the E. coli rhaBAD operon, conferring 
                     tight induction with L-rhamnose and repression with 
                     D-glucose in the presence of RhaR and RhaS (Giacalone et 
                     al., 2006)"
     CDS             2019..2042
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             2076..2096
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6837..6857
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(6878..6896)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      6892..6939
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      7037..7066
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
ORIGIN
        1 ttgagatcgt tttggtctgc gcgtaatctc ttgctctgaa aacgaaaaaa ccgccttgca
       61 gggcggtttt tcgaaggttc tctgagctac caactctttg aaccgaggta actggcttgg
      121 aggagcgcag tcaccaaaac ttgtcctttc agtttagcct taaccggcgc atgacttcaa
      181 gactaactcc tctaaatcaa ttaccagtgg ctgctgccag tggtgctttt gcatgtcttt
      241 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcggac tgaacggggg
      301 gttcgtgcat acagtccagc ttggagcgaa ctgcctaccc ggaactgagt gtcaggcgtg
      361 gaatgagaca aacgcggcca taacagcgga atgacaccgg taaaccgaaa ggcaggaaca
      421 ggagagcgca cgagggagcc gccaggggga aacgcctggt atctttatag tcctgtcggg
      481 tttcgccacc actgatttga gcgtcagatt tcgtgatgct tgtcaggggg gcggagccta
      541 tggaaaaacg gctttgccgc ggccctctca cttccctgtt aagtatcttc ctggcatctt
      601 ccaggaaatc tccgccccgt tcgtaagcca tttccgctcg ccgcagtcga acgaccgagc
      661 gtagcgagtc agtgagcgag gaagcggaat atatcctgta tcacatattc tgctgacgca
      721 ccggtgcagc cttttttctc ctgccacatg aagcacttca ctgacaccct catcagtgcc
      781 aacatagtaa gccagtatac actccgctag cgcagaaagg cccacccgaa ggtgagccag
      841 gtgattacat ttgggccctc attagaaaaa ctcatcgagc atcaaatgaa attgcaattt
      901 attcatatca ggattatcaa taccatattt ttgaaaaagc cgtttctgta atgaaggaga
      961 aaactcaccg aggcagttcc ataggatggc aagatcctgg tatcggtctg cgattccgac
     1021 tcgtccaaca tcaatacaac ctattaattt cccctcgtca aaaataaggt tatcaagtga
     1081 gaaatcacca tgagtgacga ctgaatccgg tgagaatggc aaaagtttat gcatttcttt
     1141 ccagacttgt tcaacaggcc agccattacg ctcgtcatca aaatcactcg catcaaccaa
     1201 accgttattc attcgtgatt gcgcctgagc gaggcgaaat acgcgatcgc tgttaaaagg
     1261 acaattacaa acaggaatcg agtgcaaccg gcgcaggaac actgccagcg catcaacaat
     1321 attttcacct gaatcaggat attcttctaa tacctggaac gctgtttttc cggggatcgc
     1381 agtggtgagt aaccatgcat catcaggagt acggataaaa tgcttgatgg tcggaagtgg
     1441 cataaattcc gtcagccagt ttagtctgac catctcatct gtaacatcat tggcaacgct
     1501 acctttgcca tgtttcagaa acaactctgg cgcatcgggc ttcccataca agcgatagat
     1561 tgtcgcacct gattgcccga cattatcgcg agcccattta tacccatata aatcagcatc
     1621 catgttggaa tttaatcgcg gcctcgacgt ttcccgttga atatggctca tagctcctga
     1681 aaatctcgat aactcaaaaa atacgcccgg tagtgatctt atttcattat ggtgaaagtt
     1741 ggaacctctt acgtgccgat caagaagacg gtcaaaagcc tccggtcgga ggcttttgac
     1801 tttctgctat ggaggtcagg tatgatttaa atggtcagta ttgagcgata tctagagaat
     1861 tcgtccacca caattcagca aattgtgaac atcatcacgt tcatctttcc ctggttgcca
     1921 atggcccatt ttcctgtcag taacgagaag gtcgcgaatt caggcgcttt ttagactggt
     1981 cgtaatgaaa ttctttttaa gaaggagata tacatatgca ccatcaccat catcatcatc
     2041 acaaacgtac cgcagatggt agcgaatttg aaagcccgaa aaaaaagcgt aaagttagcg
     2101 aagttgaatt tagccacgaa tactggatgc gtcatgcgtt gacgctggcg aagcgtgcgc
     2161 gtgacgaacg cgaggtgccg gtgggtgctg ttctggtgct gaacaatcgt gtaatcggtg
     2221 aaggttggaa tcgcgcgatc ggtctgcatg atccaaccgc gcacgcagaa atcatggcgc
     2281 tgcgtcaggg tggcctggtg atgcaaaatt accgtctgat cgatgcgacg ctgtatgtga
     2341 ccttcgaacc gtgtgtcatg tgtgccggtg ccatgatcca cagccgcatt ggccgtgtgg
     2401 tctttggcgt tcgtaacagc aaacgtggcg cagcaggctc tctgatgaac gtcctgaatt
     2461 acccgggtat gaaccatcgt gttgagatta cggagggcat cttagcggat gaatgtgctg
     2521 cgctgctgtg cgatttctat cgcatgccgc gtcaagtctt taacgcacag aagaaggcac
     2581 aatctagcat taactctggc ggttcgtctg gtggtagcag cggcagcgaa acgccgggta
     2641 ccagcgagag cgcaaccccg gaaagctccg gtggcagctc tggcggctct gacaaaaagt
     2701 attcgattgg cctggcgatc ggcacgaaca gcgtcggttg ggctgttatt accgacgagt
     2761 ataaagtgcc gagcaagaag tttaaagtct tgggtaacac cgatcgccat agcatcaaaa
     2821 agaatctgat tggtgccctg ctgtttgata gcggcgaaac ggcagaagcg acccgtctga
     2881 aacgtaccgc acgtcgtcgc tacactcgtc gtaaaaaccg catttgctat ttgcaagaga
     2941 tcttcagcaa tgaaatggcc aaagttgacg acagcttctt tcaccgtctg gaggagagct
     3001 ttctggttga agaggataag aaacacgagc gtcacccgat ctttggtaat atcgttgacg
     3061 aggtcgcgta tcacgagaag tacccgacga tctaccactt gcgtaagaaa ctggtggaca
     3121 gcaccgataa agccgatctg cgtctgattt acttggcgtt ggcgcacatg atcaagttcc
     3181 gcggtcattt cctgatcgag ggtgacctga acccagataa ttccgacgtc gacaagctgt
     3241 tcatccagct ggttcagacc tacaaccaat tgtttgaaga aaacccgatt aatgcatctg
     3301 gtgtcgacgc gaaggcgatt ctgagcgcgc gtctgagcaa gtctcgtcgt ctggagaatc
     3361 tgatcgcgca attgccgggt gagaagaaaa atggtttgtt tggcaacttg atcgcgctga
     3421 gcctgggtct gaccccgaac tttaagagca atttcgacct ggcagaagat gcgaaactgc
     3481 agctgagcaa agacacctat gatgacgacc tggataatct cctggcacaa attggtgacc
     3541 agtatgcgga cctgttcctg gctgcgaaaa atctgtcgga cgccattctg ctgtcggaca
     3601 ttctgcgcgt taacactgag atcacgaaag caccgctgtc tgcgagcatg attaaacgtt
     3661 atgacgagca ccatcaagac ctgaccctgc tgaaggccct ggtccgtcag caactgccgg
     3721 agaagtataa agaaattttt ttcgatcaga gcaagaacgg ttacgcaggc tatatcgatg
     3781 gcggtgcgag ccaagaagag ttttacaaat tcattaaacc gattttggag aagatggacg
     3841 gtaccgagga gttgctggtg aaactgaatc gcgaagatct gctgcgcaaa caacgtacgt
     3901 tcgataacgg tagcatccca caccagattc acctgggtga gctgcacgcg attctgcgtc
     3961 gtcaggaaga tttctaccca ttcctgaaag ataatcgtga aaagattgag aaaatcttga
     4021 cgttccgcat tccgtactac gtcggtccgc tggcacgtgg caacagccgt ttcgcttgga
     4081 tgacgcgcaa gagcgaggaa actatcacgc cgtggaactt tgaggaagtc gtcgacaaag
     4141 gtgcgagcgc gcaaagcttt attgagcgca tgaccaactt tgacaagaat ctgccgaacg
     4201 agaaagtctt gccaaagcac agcctgctgt acgagtattt cacggtgtac aatgaactga
     4261 ccaaagttaa gtatgtcacc gagggtatgc gtaaaccagc gttcctgtcc ggtgaacaga
     4321 agaaagcaat tgtggatctg ttgtttaaga cgaaccgtaa agtcacggtg aagcagttga
     4381 aagaggatta cttcaaaaag atcgaatgct ttgacagcgt ggaaattagc ggtgttgagg
     4441 accgtttcaa cgccagcttg ggcacctacc atgatctgct gaaaatcatt aaggacaagg
     4501 acttcctgga taacgaagaa aacgaggata ttctggagga tattgtcctg acgctgaccc
     4561 tcttcgaaga tcgcgaaatg attgaagaga gactgaaaac ttacgcccac ttgtttgacg
     4621 ataaagtcat gaaacaactg aagcgccgtc gttacaccgg ttggggtcgt ctgagccgta
     4681 agctgatcaa cggtatccgt gataagcaat ctggcaaaac gatcctggac tttctgaaaa
     4741 gcgatggttt cgcgaaccgc aacttcatgc agcttattca cgacgattct ctgaccttta
     4801 aggaggacat ccaaaaagcc caggttagcg gtcagggcga ttccctgcat gagcatattg
     4861 ctaacttagc gggtagcccg gcaatcaaaa agggcattct gcaaaccgtt aaggttgtgg
     4921 acgagttggt taaggtcatg ggccgtcata agccggagaa tatcgttatt gagatggcgc
     4981 gcgagaatca gaccacgcag aaaggccaaa agaatagccg tgaacgtatg aaacgtattg
     5041 aagagggtat caaagagctg ggcagccaaa tcttgaagga acatccggtg gagaacacgc
     5101 agctgcagaa tgaaaagctg tacctgtact atctgcaaaa cggtcgtgat atgtacgttg
     5161 accaagagct ggacatcaac cgcctgagcg attatgatgt tgatcacatt gtgccgcaat
     5221 cttttctgaa ggacgacagc atcgacaaca aagttttgac ccgtagcgac aagaatcgcg
     5281 gtaaaagcga taatgttccg agcgaagagg tcgttaagaa gatgaaaaac tattggcgtc
     5341 agttgctgaa tgcaaaactg attacccaac gcaaattcga caatttgact aaagcggagc
     5401 gtggtggtct gagcgagctg gacaaggcgg gtttcattaa gcgtcaactg gttgaaactc
     5461 gtcagatcac gaagcacgtg gcgcagattc tggacagccg catgaatact aagtacgatg
     5521 aaaatgataa actcattcgt gaggtcaagg tcattaccct gaagtctaag ctcgtcagcg
     5581 attttcgtaa agacttccag ttctataagg tgcgcgaaat caacaattac caccacgccc
     5641 atgatgcata tctgaacgcc gtggtgggta cggctctgat caagaaatat ccgaaactgg
     5701 agtccgagtt tgtctatggt gactacaaag tgtatgacgt ccgcaaaatg atcgccaaat
     5761 cggaacaaga gattggcaag gcaaccgcga agtacttttt ctattccaac attatgaact
     5821 ttttcaagac cgagatcacc ctggcaaacg gtgagatccg taaacgtccg ctgatcgaaa
     5881 ctaacggtga aaccggtgaa atcgtgtggg acaaaggtcg tgactttgca accgtgcgta
     5941 aagtgctgtc gatgccacag gtcaacatcg ttaagaaaac cgaagttcaa actggtggct
     6001 ttagcaagga gagcattcgt cctaagcgca acagcgacaa gctgattgca cgtaagaagg
     6061 actgggaccc gaagaagtac ggcggctttg tgtccccgac ggttgcgtat agcgttctcg
     6121 ttgtggcgaa agtcgagaaa ggcaagagca aaaagctgaa aagcgttaaa gaattgctgg
     6181 gtatcacgat tatggagcgt agcagcttcg aaaagaatcc tattgatttc ttggaagcaa
     6241 aaggttacaa ggaggtgaag aaagacttga tcatcaaact gccgaagtat tccctgttcg
     6301 agctggaaaa cggccgcaaa cgtatgctgg ccagcgctcg ctttctgcaa aagggcaacg
     6361 aactggctct gccgagcaaa tatgtgaatt tcctgtacct ggccagccac tacgagaaac
     6421 tgaagggctc cccggaagat aacgagcaga agcagctgtt cgttgagcag cacaagcatt
     6481 acctggacga gattatcgag cagatcagcg agtttagcaa acgcgtgatt ctggcggatg
     6541 cgaatttgga taaagttctg tcggcctata ataaacaccg cgacaaaccg attcgcgagc
     6601 aggcggagaa cattatccac ttgttcaccc tgaccaatct gggtgctccg cgtgctttca
     6661 aatactttga taccacgatt gatcgcaaag tgtatcgttc caccaaagag gtactggacg
     6721 cgaccctgat ccaccagagc atcaccggct tgtacgaaac gcgcatcgat ctgtctcaac
     6781 tgggcggtga cagcggtggt tcgaaacgta ccgcggatgg ctctgaattt gaacctccga
     6841 aaaagaaacg caaggtgtaa ggttagagcg gccgccaccg ctgagcaata actagcataa
     6901 ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc
     6961 gggtaacgaa ttcaagcttg atatcattca ggacgagcct cagactccag cgtaactgga
     7021 ctgcaatcaa ctcactggct caccttcacg ggtgggcctt tcttcggtag aagtcttctt
     7081 aataagatga tcttc
//
