LOCUS       sequence-334014-        7095 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For bacterial purification of ABE8e-NRCH protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7095)
  TITLE     ABE8e-NRCH for bacterial and human expression
  JOURNAL   Unpublished
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: 
            "Unpublished"
COMMENT     Sequence Label: pRha-ABE8e-NRCH
FEATURES             Location/Qualifiers
     source          1..7095
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2724..2744
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(2765..2783)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2779..2826
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2924..2953
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      2983..3528
                     /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     3429..3448
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3645..3662
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3793..3820)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3844..4653)
                     /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="MSNIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     4567..4586
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(4654..4744)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(4753..4783)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        4848..4966
                     /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             5001..5024
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             5058..5078
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 ggaactttga ggaagtcgtc gacaaaggtg cgagcgcgca aagctttatt gagcgcatga
       61 ccaactttga caagaatctg ccgaacgaga aagtcttgcc aaagcacagc ctgctgtacg
      121 agtatttcac ggtgtacaat gaactgacca aagttaagta tgtcaccgag ggtatgcgta
      181 aaccagcgtt cctgtccggt gaacagaaga aagcaattgt ggatctgttg tttaagacga
      241 accgtaaagt cacggtgaag cagttgaaag aggattactt caaaaagatc gaatgctttg
      301 acagcgtgga aattagcggt gttgaggacc gtttcaacgc cagcttgggc acctaccatg
      361 atctgctgaa aatcattaag gacaaggact tcctggataa cgaagaaaac gaggatattc
      421 tggaggatat tgtcctgacg ctgaccctct tcgaagatcg cgaaatgatt gaagagagac
      481 tgaaaactta cgcccacttg tttgacgata aagtcatgaa acaactgaag cgcctgcgtt
      541 acaccggttg gggtcgtctg agccgtaagc tgatcaacgg tatccgtgat aagcaatctg
      601 gcaaaacgat cctggacttt ctgaaaagcg atggtttcgc gaaccgcaac ttcatgcagc
      661 ttattcacga cgattctctg acctttaagg aggacatcca aaaagcccag gttagcggtc
      721 agggcgattc cctgcatgag catattgcta acttagcggg tagcccggca atcaaaaagg
      781 gcattctgca aaccgttaag gttgtggacg agttggttaa ggtcatgggc ggccataagc
      841 cggagaatat cgttattgag atggcgcgcg agaatcagac cacgcagaaa ggccaaaaga
      901 atagccgtga acgtatgaaa cgtattgaag agggtatcaa agagctgggc agccaaatct
      961 tgaaggaaca tccggtggag aacacgcagc tgcagaatga aaagctgtac ctgtactatc
     1021 tgcaaaacgg tcgtgatatg tacgttgacc aagagctgga catcaaccgc ctgagcgatt
     1081 atgatgttga tcacattgtg ccgcaatctt ttctgaagga cgacagcatc gacaacaaag
     1141 ttttgacccg tagcgacaag aatcgcggta aaagcgataa tgttccgagc gaagaggtcg
     1201 ttaagaagat gaaaaactat tggcgtcagt tgctgaatgc aaaactgatt acccaacgca
     1261 aattcgacaa tttgactaaa gcggagcgtg gtggtctgag cgagctggac aaggcgggtt
     1321 tcattaagcg tcaactggtt gaaactcgtc agatcacgaa gcacgtggcg cagattctgg
     1381 acagccgcat gaatactaag tacgatgaaa atgataaact cattcgtgag gtcaaggtca
     1441 ttaccctgaa gtctaagctc gtcagcgatt ttcgtaaaga cttccagttc tataaggtgc
     1501 gcgaaatcaa caattaccac cacgcccatg atgcatatct gaacgccgtg gtgggtacgg
     1561 ctctgatcaa gaaatatccg aaactggagt ccgagtttgt ctatggtgac tacaaagtgt
     1621 atgacgtccg caaaatgatc gccaaatcgg aacaagagat tggcaaggca accgcgaagt
     1681 actttttcta ttccaacatt atgaactttt tcaagaccga gatcaccctg gcaaacggtg
     1741 agatccgtaa acgtccgctg atcgaaacta acggtgaaac cggtgaaatc gtgtgggaca
     1801 aaggtcgtga ctttgcaacc gtgcgtaaag tgctgtcgat gccacaggtc aacatcgtta
     1861 agaaaaccga agttcaaact ggtggcttta gcaaggagag cattctgcct aagggtaaca
     1921 gcgacaagct gattgcacgt aagaaggact gggacccgaa gaagtacggc ggctttaact
     1981 ccccgacggt tgcgtatagc gttctcgttg tggcgaaagt cgagaaaggc aagagcaaaa
     2041 agctgaaaag cgttaaagaa ttgctgggta tcacgattat ggagcgtagc agcttcgaaa
     2101 agaatcctat tgatttcttg gaagcaaaag gttacaagga ggtgaagaaa gacttgatca
     2161 tcaaactgcc gaagtattcc ctgttcgagc tggaaaacgg ccgcaaacgt atgctggcca
     2221 gcgctggtgt gctgcaaaag ggcaacgaac tggctctgcc gagcaaatat gtgaatttcc
     2281 tgtacctggc cagccactac gagaaactga agggctcccc ggaagataac gagcagaagc
     2341 agctgttcgt tgagcagcac aagcattacc tggacgagat tatcgagcag atcagcgagt
     2401 ttagcaaacg cgtgattctg gcggatgcga atttggataa agttctgtcg gcctataata
     2461 aacaccgcga caaaccgatt cgcgagcagg cggagaacat tatccacttg ttcaccctga
     2521 ccaatctggg tgctccggct gctttcaaat actttgatac cacgattaac cgcaaacagt
     2581 ataacaccac caaagaggta ctggacgcga ccctgatccg ccagagcatc accggcttgt
     2641 acgaaacgcg catcgatctg tctcaactgg gcggtgacag cggtggttcg aaacgcacag
     2701 ccgatggatc agaatttgaa tctccgaaaa agaaacgcaa ggtgtaaggt tagagcggcc
     2761 gccaccgctg agcaataact agcataaccc cttggggcct ctaaacgggt cttgaggggt
     2821 tttttgctga aaggaggaac tatatccggg taacgaattc aagcttgata tcattcagga
     2881 cgagcctcag actccagcgt aactggactg caatcaactc actggctcac cttcacgggt
     2941 gggcctttct tcggtagaag tcttcttaat aagatgatct tcttgagatc gttttggtct
     3001 gcgcgtaatc tcttgctctg aaaacgaaaa aaccgccttg cagggcggtt tttcgaaggt
     3061 tctctgagct accaactctt tgaaccgagg taactggctt ggaggagcgc agtcaccaaa
     3121 acttgtcctt tcagtttagc cttaaccggc gcatgacttc aagactaact cctctaaatc
     3181 aattaccagt ggctgctgcc agtggtgctt ttgcatgtct ttccgggttg gactcaagac
     3241 gatagttacc ggataaggcg cagcggtcgg actgaacggg gggttcgtgc atacagtcca
     3301 gcttggagcg aactgcctac ccggaactga gtgtcaggcg tggaatgaga caaacgcggc
     3361 cataacagcg gaatgacacc ggtaaaccga aaggcaggaa caggagagcg cacgagggag
     3421 ccgccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca ccactgattt
     3481 gagcgtcaga tttcgtgatg cttgtcaggg gggcggagcc tatggaaaaa cggctttgcc
     3541 gcggccctct cacttccctg ttaagtatct tcctggcatc ttccaggaaa tctccgcccc
     3601 gttcgtaagc catttccgct cgccgcagtc gaacgaccga gcgtagcgag tcagtgagcg
     3661 aggaagcgga atatatcctg tatcacatat tctgctgacg caccggtgca gccttttttc
     3721 tcctgccaca tgaagcactt cactgacacc ctcatcagtg ccaacatagt aagccagtat
     3781 acactccgct agcgcagaaa ggcccacccg aaggtgagcc aggtgattac atttgggccc
     3841 tcattagaaa aactcatcga gcatcaaatg aaattgcaat ttattcatat caggattatc
     3901 aataccatat ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt
     3961 ccataggatg gcaagatcct ggtatcggtc tgcgattccg actcgtccaa catcaataca
     4021 acctattaat ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac
     4081 gactgaatcc ggtgagaatg gcaaaagttt atgcatttct ttccagactt gttcaacagg
     4141 ccagccatta cgctcgtcat caaaatcact cgcatcaacc aaaccgttat tcattcgtga
     4201 ttgcgcctga gcgaggcgaa atacgcgatc gctgttaaaa ggacaattac aaacaggaat
     4261 cgagtgcaac cggcgcagga acactgccag cgcatcaaca atattttcac ctgaatcagg
     4321 atattcttct aatacctgga acgctgtttt tccggggatc gcagtggtga gtaaccatgc
     4381 atcatcagga gtacggataa aatgcttgat ggtcggaagt ggcataaatt ccgtcagcca
     4441 gtttagtctg accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag
     4501 aaacaactct ggcgcatcgg gcttcccata caagcgatag attgtcgcac ctgattgccc
     4561 gacattatcg cgagcccatt tatacccata taaatcagca tccatgttgg aatttaatcg
     4621 cggcctcgac gtttcccgtt gaatattgct catagctcct gaaaatctcg ataactcaaa
     4681 aaatacgccc ggtagtgatc ttatttcatt atggtgaaag ttggaacctc ttacgtgccg
     4741 atcaagaaga cggtcaaaag cctccggtcg gaggcttttg actttctgct atggaggtca
     4801 ggtatgattt aaatggtcag tattgagcga tatctagaga attcgtccac cacaattcag
     4861 caaattgtga acatcatcac gttcatcttt ccctggttgc caatggccca ttttcctgtc
     4921 agtaacgaga aggtcgcgaa ttcaggcgct ttttagactg gtcgtaatga aattcttttt
     4981 aagaaggaga tatacatatg caccatcacc atcatcatca tcacaaacgt accgcagatg
     5041 gtagcgaatt tgaaagcccg aaaaaaaagc gtaaagttag cgaagttgaa tttagccacg
     5101 aatactggat gcgtcatgcg ttgacgctgg cgaagcgtgc gcgtgacgaa cgcgaggtgc
     5161 cggtgggtgc tgttctggtg ctgaacaatc gtgtaatcgg tgaaggttgg aatcgcgcga
     5221 tcggtctgca tgatccaacc gcgcacgcag aaatcatggc gctgcgtcag ggtggcctgg
     5281 tgatgcaaaa ttaccgtctg atcgatgcga cgctgtatgt gaccttcgaa ccgtgtgtca
     5341 tgtgtgccgg tgccatgatc cacagccgca ttggccgtgt ggtctttggc gttcgtaaca
     5401 gcaaacgtgg cgcagcaggc tctctgatga acgtcctgaa ttacccgggt atgaaccatc
     5461 gtgttgagat tacggagggc atcttagcgg atgaatgtgc tgcgctgctg tgcgatttct
     5521 atcgcatgcc gcgtcaagtc tttaacgcac agaagaaggc acaatctagc attaactctg
     5581 gcggttcgtc tggtggtagc agcggcagcg aaacgccggg taccagcgag agcgcaaccc
     5641 cggaaagctc cggtggcagc tctggcggct ctgacaaaaa gtattcgatt ggcctgacca
     5701 tcggcacgaa cagcgtcggt tgggctgtta ttaccgacga gtataaagtg ccgagcaaga
     5761 agtttaaagt cttgggtaac accgatcgcc atagcatcaa aaagaatctg attggtgccc
     5821 tgctgtttga tagcggcgaa acggcagaag cgacccgtct gaaacgtacc gcacgtcgtc
     5881 gctacactcg tcgtaaaaac cgcatttgct atttgcaaga gatcttcagc aatgaaatgg
     5941 ccaaagttga cgacagcttc tttcaccgtc tggaggagag ctttctggtt gaagaggata
     6001 agaaacacga gcgtcacccg atctttggta atatcgttga cgaggtcgcg tatcacgaga
     6061 agtacccgac gatctaccac ttgcgtaaga aactggtgga cagcaccgat aaagccgatc
     6121 tgcgtctgat ttacttggcg ttggcgcaca tgatcaagtt ccgcggtcat ttcctgatcg
     6181 agggtgacct gaacccagat aattccgacg tcgacaagct gttcatccag ctggttcaga
     6241 cctacaacca attgtttgaa gaaaacccga ttaatgcatc tggtgtcgac gcgaaggcga
     6301 ttctgagcgc gcgtctgagc aagtctcgtc gtctggagaa tctgatcgcg caattgccgg
     6361 gtgagaagaa aaatggtttg tttggcaact tgatcgcgct gagcctgggt ctgaccccga
     6421 actttaagag caatttcgac ctggcagaag atgcgaaact gcagctgagc aaagacacct
     6481 atgatgacga cctggataat ctcctggcac aaattggtga ccagtatgcg gacctgttcc
     6541 tggctgcgaa aaatctgtcg gacgccattc tgctgtcgga cattctgcgc gttaacactg
     6601 agatcacgaa agcaccgctg tctgcgagca tggtgaaacg ttatgacgag caccatcaag
     6661 acctgaccct gctgaaggcc ctggtccgtc agcaactgcc ggagaagtat aaagaaattt
     6721 ttttcgatca gagcaagaac ggttacgcag gctatatcga tggcggtgcg agccaagaag
     6781 agttttacaa attcattaaa ccgattttgg agaagatgga cggtaccgag gagttgctgg
     6841 tgaaactgaa tcgcgaagat ctgctgcgca aacaacgtac gttcgataac ggtattatcc
     6901 cacaccagat tcacctgggt gagctgcacg cgattctgcg tcgtcagggc gatttctacc
     6961 cattcctgaa agataatcgt gaaaagattg agaaaatctt gacgttccgc attccgtact
     7021 acgtcggtcc gctggcacgt ggcaacagcc gtttcgcttg gatgacgcgc aagagcgagg
     7081 aaactatcac gccgt
//
