LOCUS       sequence-334867-        7707 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the base editor ABE7.10-SpCas9-NG in bacterial cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7707)
  AUTHORS   Krishnamurthy S, Traore S, Cooney AL, Brommel CM, Kulhankova K, Sinn
            PL, Newby GA, Liu DR, McCray PB
  TITLE     Functional correction of CFTR mutations in human airway epithelial 
            cells using adenine base editors.
  JOURNAL   Nucleic Acids Res. 2021 Oct 11;49(18):10558-10572. doi: 
            10.1093/nar/gkab788.
  PUBMED    34520545
REFERENCE   2  (bases 1 to 7707)
  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"; doi: "10"; journalName: 
            "Nucleic Acids Res"; date: "2021-10-11- 11"; volume: "49"; issue: 
            "18"; pages: "10558-10572"
COMMENT     Sequence Label: pRha-ABE7.10-SpCas9-NG
FEATURES             Location/Qualifiers
     source          1..7707
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(35..125)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(134..164)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        229..347
                     /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             382..405
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             439..459
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5809..5829
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(5853..5871)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5867..5914
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      6012..6041
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      6071..6616
                     /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     6517..6536
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6733..6750
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(6881..6908)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     7655..7674
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 gcggcctcga cgtttcccgt tgaatattgc tcatagctcc tgaaaatctc gataactcaa
       61 aaaatacgcc cggtagtgat cttatttcat tatggtgaaa gttggaacct cttacgtgcc
      121 gatcaagaag acggtcaaaa gcctccggtc ggaggctttt gactttctgc tatggaggtc
      181 aggtatgatt taaatggtca gtattgagcg atatctagag aattcgtcca ccacaattca
      241 gcaaattgtg aacatcatca cgttcatctt tccctggttg ccaatggccc attttcctgt
      301 cagtaacgag aaggtcgcga attcaggcgc tttttagact ggtcgtaatg aaattctttt
      361 taagaaggag atatacatat gcaccatcac catcatcatc atcacaaacg taccgcagat
      421 ggtagcgaat ttgaaagccc gaaaaaaaag cgtaaagttg gttcaggcgg ctcgggttcc
      481 gaagtggaat tttcccacga gtactggatg agacatgcgc tgaccctggc gaaacgtgcg
      541 tgggatgaac gtgaggttcc ggtaggcgca gtcttggtgc ataacaatcg cgtgattggt
      601 gagggctgga atcgtccgat cggtcgccac gatccgacgg cacacgcaga gattatggcc
      661 ctgcgccagg gtggtttggt tatgcagaat taccgcttga ttgatgccac cctgtacgtc
      721 accctggagc cgtgtgtgat gtgcgctggt gcaatgatcc actcccgtat tggccgcgtt
      781 gtgtttggtg cccgcgacgc taaaaccggt gcagcgggct ctttgatgga cgtgttgcac
      841 caccctggta tgaatcatcg tgtggagatt accgaaggca tcctggcgga tgagtgcgcg
      901 gcactgctga gcgacttttt ccgtatgcgc cgtcaggaga tcaaagctca aaagaaggcc
      961 cagagcagca ccgactcggg tggttctagc ggtggcagca gcggtagcga aacccctggc
     1021 acctccgagt ccgctacccc ggagtctagc ggcggttctt ccggcggttc cagcgaagtt
     1081 gaatttagcc acgaatactg gatgcgtcat gcgttgacgc tggcgaagcg tgcgcgtgac
     1141 gaacgcgagg tgccggtggg tgctgttctg gtgctgaaca atcgtgtaat cggtgaaggt
     1201 tggaatcgcg cgatcggtct gcatgatcca accgcgcacg cagaaatcat ggcgctgcgt
     1261 cagggtggcc tggtgatgca aaattaccgt ctgatcgatg cgacgctgta tgtgaccttc
     1321 gaaccgtgtg tcatgtgtgc cggtgccatg atccacagcc gcattggccg tgtggtcttt
     1381 ggcgttcgta acgcgaaaac cggcgcagca ggctctctga tggatgtcct gcattacccg
     1441 ggtatgaacc atcgtgttga gattacggag ggcatcttag cggatgaatg tgctgcgctg
     1501 ctgtgctatt tcttccgcat gccgcgtcaa gtctttaacg cacagaagaa ggcacaatct
     1561 agcacggact ctggcggttc gtctggtggt agcagcggca gcgaaacgcc gggtaccagc
     1621 gagagcgcaa ccccggaaag ctccggtggc agctctggcg gctctgacaa aaagtattcg
     1681 attggcctgg cgatcggcac gaacagcgtc ggttgggctg ttattaccga cgagtataaa
     1741 gtgccgagca agaagtttaa agtcttgggt aacaccgatc gccatagcat caaaaagaat
     1801 ctgattggtg ccctgctgtt tgatagcggc gaaacggcag aagcgacccg tctgaaacgt
     1861 accgcacgtc gtcgctacac tcgtcgtaaa aaccgcattt gctatttgca agagatcttc
     1921 agcaatgaaa tggccaaagt tgacgacagc ttctttcacc gtctggagga gagctttctg
     1981 gttgaagagg ataagaaaca cgagcgtcac ccgatctttg gtaatatcgt tgacgaggtc
     2041 gcgtatcacg agaagtaccc gacgatctac cacttgcgta agaaactggt ggacagcacc
     2101 gataaagccg atctgcgtct gatttacttg gcgttggcgc acatgatcaa gttccgcggt
     2161 catttcctga tcgagggtga cctgaaccca gataattccg acgtcgacaa gctgttcatc
     2221 cagctggttc agacctacaa ccaattgttt gaagaaaacc cgattaatgc atctggtgtc
     2281 gacgcgaagg cgattctgag cgcgcgtctg agcaagtctc gtcgtctgga gaatctgatc
     2341 gcgcaattgc cgggtgagaa gaaaaatggt ttgtttggca acttgatcgc gctgagcctg
     2401 ggtctgaccc cgaactttaa gagcaatttc gacctggcag aagatgcgaa actgcagctg
     2461 agcaaagaca cctatgatga cgacctggat aatctcctgg cacaaattgg tgaccagtat
     2521 gcggacctgt tcctggctgc gaaaaatctg tcggacgcca ttctgctgtc ggacattctg
     2581 cgcgttaaca ctgagatcac gaaagcaccg ctgtctgcga gcatgattaa acgttatgac
     2641 gagcaccatc aagacctgac cctgctgaag gccctggtcc gtcagcaact gccggagaag
     2701 tataaagaaa tttttttcga tcagagcaag aacggttacg caggctatat cgatggcggt
     2761 gcgagccaag aagagtttta caaattcatt aaaccgattt tggagaagat ggacggtacc
     2821 gaggagttgc tggtgaaact gaatcgcgaa gatctgctgc gcaaacaacg tacgttcgat
     2881 aacggtagca tcccacacca gattcacctg ggtgagctgc acgcgattct gcgtcgtcag
     2941 gaagatttct acccattcct gaaagataat cgtgaaaaga ttgagaaaat cttgacgttc
     3001 cgcattccgt actacgtcgg tccgctggca cgtggcaaca gccgtttcgc ttggatgacg
     3061 cgcaagagcg aggaaactat cacgccgtgg aactttgagg aagtcgtcga caaaggtgcg
     3121 agcgcgcaaa gctttattga gcgcatgacc aactttgaca agaatctgcc gaacgagaaa
     3181 gtcttgccaa agcacagcct gctgtacgag tatttcacgg tgtacaatga actgaccaaa
     3241 gttaagtatg tcaccgaggg tatgcgtaaa ccagcgttcc tgtccggtga acagaagaaa
     3301 gcaattgtgg atctgttgtt taagacgaac cgtaaagtca cggtgaagca gttgaaagag
     3361 gattacttca aaaagatcga atgctttgac agcgtggaaa ttagcggtgt tgaggaccgt
     3421 ttcaacgcca gcttgggcac ctaccatgat ctgctgaaaa tcattaagga caaggacttc
     3481 ctggataacg aagaaaacga ggatattctg gaggatattg tcctgacgct gaccctcttc
     3541 gaagatcgcg aaatgattga agagagactg aaaacttacg cccacttgtt tgacgataaa
     3601 gtcatgaaac aactgaagcg ccgtcgttac accggttggg gtcgtctgag ccgtaagctg
     3661 atcaacggta tccgtgataa gcaatctggc aaaacgatcc tggactttct gaaaagcgat
     3721 ggtttcgcga accgcaactt catgcagctt attcacgacg attctctgac ctttaaggag
     3781 gacatccaaa aagcccaggt tagcggtcag ggcgattccc tgcatgagca tattgctaac
     3841 ttagcgggta gcccggcaat caaaaagggc attctgcaaa ccgttaaggt tgtggacgag
     3901 ttggttaagg tcatgggccg tcataagccg gagaatatcg ttattgagat ggcgcgcgag
     3961 aatcagacca cgcagaaagg ccaaaagaat agccgtgaac gtatgaaacg tattgaagag
     4021 ggtatcaaag agctgggcag ccaaatcttg aaggaacatc cggtggagaa cacgcagctg
     4081 cagaatgaaa agctgtacct gtactatctg caaaacggtc gtgatatgta cgttgaccaa
     4141 gagctggaca tcaaccgcct gagcgattat gatgttgatc acattgtgcc gcaatctttt
     4201 ctgaaggacg acagcatcga caacaaagtt ttgacccgta gcgacaagaa tcgcggtaaa
     4261 agcgataatg ttccgagcga agaggtcgtt aagaagatga aaaactattg gcgtcagttg
     4321 ctgaatgcaa aactgattac ccaacgcaaa ttcgacaatt tgactaaagc ggagcgtggt
     4381 ggtctgagcg agctggacaa ggcgggtttc attaagcgtc aactggttga aactcgtcag
     4441 atcacgaagc acgtggcgca gattctggac agccgcatga atactaagta cgatgaaaat
     4501 gataaactca ttcgtgaggt caaggtcatt accctgaagt ctaagctcgt cagcgatttt
     4561 cgtaaagact tccagttcta taaggtgcgc gaaatcaaca attaccacca cgcccatgat
     4621 gcatatctga acgccgtggt gggtacggct ctgatcaaga aatatccgaa actggagtcc
     4681 gagtttgtct atggtgacta caaagtgtat gacgtccgca aaatgatcgc caaatcggaa
     4741 caagagattg gcaaggcaac cgcgaagtac tttttctatt ccaacattat gaactttttc
     4801 aagaccgaga tcaccctggc aaacggtgag atccgtaaac gtccgctgat cgaaactaac
     4861 ggtgaaaccg gtgaaatcgt gtgggacaaa ggtcgtgact ttgcaaccgt gcgtaaagtg
     4921 ctgtcgatgc cacaggtcaa catcgttaag aaaaccgaag ttcaaactgg tggctttagc
     4981 aaggagagca ttcgtcctaa gcgcaacagc gacaagctga ttgcacgtaa gaaggactgg
     5041 gacccgaaga agtacggcgg ctttgtgtcc ccgacggttg cgtatagcgt tctcgttgtg
     5101 gcgaaagtcg agaaaggcaa gagcaaaaag ctgaaaagcg ttaaagaatt gctgggtatc
     5161 acgattatgg agcgtagcag cttcgaaaag aatcctattg atttcttgga agcaaaaggt
     5221 tacaaggagg tgaagaaaga cttgatcatc aaactgccga agtattccct gttcgagctg
     5281 gaaaacggcc gcaaacgtat gctggccagc gctcgctttc tgcaaaaggg caacgaactg
     5341 gctctgccga gcaaatatgt gaatttcctg tacctggcca gccactacga gaaactgaag
     5401 ggctccccgg aagataacga gcagaagcag ctgttcgttg agcagcacaa gcattacctg
     5461 gacgagatta tcgagcagat cagcgagttt agcaaacgcg tgattctggc ggatgcgaat
     5521 ttggataaag ttctgtcggc ctataataaa caccgcgaca aaccgattcg cgagcaggcg
     5581 gagaacatta tccacttgtt caccctgacc aatctgggtg ctccgcgtgc tttcaaatac
     5641 tttgatacca cgattgatcg caaagtgtat cgttccacca aagaggtact ggacgcgacc
     5701 ctgatccacc agagcatcac cggcttgtac gaaacgcgca tcgatctgtc tcaactgggc
     5761 ggtgacagcg gtggttcgaa acgtaccgcg gatggctctg aatttgaacc gaagaagaaa
     5821 cgcaaagtct aataaggtta gagcggccgc caccgctgag caataactag cataacccct
     5881 tggggcctct aaacgggtct tgaggggttt tttgctgaaa ggaggaacta tatccgggta
     5941 acgaattcaa gcttgatatc attcaggacg agcctcagac tccagcgtaa ctggactgca
     6001 atcaactcac tggctcacct tcacgggtgg gcctttcttc ggtagaagtc ttcttaataa
     6061 gatgatcttc ttgagatcgt tttggtctgc gcgtaatctc ttgctctgaa aacgaaaaaa
     6121 ccgccttgca gggcggtttt tcgaaggttc tctgagctac caactctttg aaccgaggta
     6181 actggcttgg aggagcgcag tcaccaaaac ttgtcctttc agtttagcct taaccggcgc
     6241 atgacttcaa gactaactcc tctaaatcaa ttaccagtgg ctgctgccag tggtgctttt
     6301 gcatgtcttt ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcggac
     6361 tgaacggggg gttcgtgcat acagtccagc ttggagcgaa ctgcctaccc ggaactgagt
     6421 gtcaggcgtg gaatgagaca aacgcggcca taacagcgga atgacaccgg taaaccgaaa
     6481 ggcaggaaca ggagagcgca cgagggagcc gccaggggga aacgcctggt atctttatag
     6541 tcctgtcggg tttcgccacc actgatttga gcgtcagatt tcgtgatgct tgtcaggggg
     6601 gcggagccta tggaaaaacg gctttgccgc ggccctctca cttccctgtt aagtatcttc
     6661 ctggcatctt ccaggaaatc tccgccccgt tcgtaagcca tttccgctcg ccgcagtcga
     6721 acgaccgagc gtagcgagtc agtgagcgag gaagcggaat atatcctgta tcacatattc
     6781 tgctgacgca ccggtgcagc cttttttctc ctgccacatg aagcacttca ctgacaccct
     6841 catcagtgcc aacatagtaa gccagtatac actccgctag cgcagaaagg cccacccgaa
     6901 ggtgagccag gtgattacat ttgggccctc attagaaaaa ctcatcgagc atcaaatgaa
     6961 attgcaattt attcatatca ggattatcaa taccatattt ttgaaaaagc cgtttctgta
     7021 atgaaggaga aaactcaccg aggcagttcc ataggatggc aagatcctgg tatcggtctg
     7081 cgattccgac tcgtccaaca tcaatacaac ctattaattt cccctcgtca aaaataaggt
     7141 tatcaagtga gaaatcacca tgagtgacga ctgaatccgg tgagaatggc aaaagtttat
     7201 gcatttcttt ccagacttgt tcaacaggcc agccattacg ctcgtcatca aaatcactcg
     7261 catcaaccaa accgttattc attcgtgatt gcgcctgagc gaggcgaaat acgcgatcgc
     7321 tgttaaaagg acaattacaa acaggaatcg agtgcaaccg gcgcaggaac actgccagcg
     7381 catcaacaat attttcacct gaatcaggat attcttctaa tacctggaac gctgtttttc
     7441 cggggatcgc agtggtgagt aaccatgcat catcaggagt acggataaaa tgcttgatgg
     7501 tcggaagtgg cataaattcc gtcagccagt ttagtctgac catctcatct gtaacatcat
     7561 tggcaacgct acctttgcca tgtttcagaa acaactctgg cgcatcgggc ttcccataca
     7621 agcgatagat tgtcgcacct gattgcccga cattatcgcg agcccattta tacccatata
     7681 aatcagcatc catgttggaa tttaatc
//
