LOCUS       sequence-258758-        7879 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid bearing APOBEC1 coding sequence. In presence of its cofactor
            and overexpressing an artificial reporter with editable CAA codon in
            APOB sequence we can visualize its editing activity.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7879)
  AUTHORS   Chieca M, Montini M, Severi F, Pecori R, Conticello S
  TITLE     Dimerisation of APOBEC1 is dispensable for its RNA editing activity
  JOURNAL   bioRxiv 2021
REFERENCE   2  (bases 1 to 7879)
  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: "bioRxiv 
            2021"
COMMENT     Sequence Label: pBlueScript-humanAPOBEC1
FEATURES             Location/Qualifiers
     source          1..7879
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(33..49)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(34..50)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(79..97)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(118..134)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(118..134)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(131..153)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    142..158
                     /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(166..172,173..190,191..196))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    211..232
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(349..366)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(520..1108)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(600..619)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1279..2070,2071..2139))
                     /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     1902..1921
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2140..2244)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2270..2725)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2407..2428)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2619..2638
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2852..2874
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2866..2883
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2867..2883
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2890..2909
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2890..2908
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     2941..2957
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2941..2957
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     protein_bind    complement(2960..2993)
                     /label=lox71
                     /bound_moiety="Cre recombinase"
                     /note="Left element (LE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3011..3286
                     /label=chicken beta-actin promoter
     intron          3287..4299
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     polyA_signal    5282..5416
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        5434..5709
                     /label=chicken beta-actin promoter
     intron          5710..6722
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     CDS             6873..7472
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(6873..6892)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7369..7389
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    7702..7836
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    complement(7838..7871)
                     /label=lox66
                     /bound_moiety="Cre recombinase"
                     /note="Right element (RE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 cccgggctgc aggaattcga tatcaagctt atcgataccg tcgacctcga gggggggccc
       61 ggtacccagc ttttgttccc tttagtgagg gttaatttcg agcttggcgt aatcatggtc
      121 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
      181 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
      241 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
      301 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
      361 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
      421 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
      481 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
      541 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
      601 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
      661 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
      721 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
      781 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
      841 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
      901 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
      961 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     1021 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     1081 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     1141 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     1201 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     1261 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     1321 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     1381 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     1441 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     1501 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     1561 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     1621 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     1681 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     1741 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     1801 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     1861 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     1921 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     1981 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     2041 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     2101 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     2161 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     2221 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccaccta aattgtaagc
     2281 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     2341 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     2401 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     2461 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     2521 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     2581 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     2641 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     2701 cttaatgcgc cgctacaggg cgcgtcccat tcgccattca ggctgcgcaa ctgttgggaa
     2761 gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca
     2821 aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc
     2881 agtgaattgt aatacgactc actatagggc gaattggagc tccaccgcgg tggcggccgc
     2941 tctagaacta gtggatccct accgttcgta tagcatacat tatacgaagt tattgatcag
     3001 ggctagtggc tcgaggtgag ccccacgttc tgcttcactc tccccatctc ccccccctcc
     3061 ccacccccaa ttttgtattt atttattttt taattatttt gtgcagcgat gggggcgggg
     3121 gggggggggg cgcgcgccag gcggggcggg gcggggcgag gggcggggcg gggcgaggcg
     3181 gagaggtgcg gcggcagcca atcagagcgg cgcgctccga aagtttcctt ttatggcgag
     3241 gcggcggcgg cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag tcgctgcgtt
     3301 gccttcgccc cgtgccccgc tccgcgccgc ctcgcgccgc ccgccccggc tctgactgac
     3361 cgcgttactc ccacaggtga gcgggcggga cggcccttct cctccgggct gtaattagcg
     3421 cttggtttaa tgacggctcg tttcttttct gtggctgcgt gaaagcctta aagggctccg
     3481 ggagggccct ttgtgcgggg gggagcggct cggggggtgc gtgcgtgtgt gtgtgcgtgg
     3541 ggagcgccgc gtgcggcccg cgctgcccgg cggctgtgag cgctgcgggc gcggcgcggg
     3601 gctttgtgcg ctccgcgtgt gcgcgagggg agcgcggccg ggggcggtgc cccgcggtgc
     3661 gggggggctg cgaggggaac aaaggctgcg tgcggggtgt gtgcgtgggg gggtgagcag
     3721 ggggtgtggg cgcggcggtc gggctgtaac ccccccctgc acccccctcc ccgagttgct
     3781 gagcacggcc cggcttcggg tgcggggctc cgtgcggggc gtggcgcggg gctcgccgtg
     3841 ccgggcgggg ggtggcggca ggtgggggtg ccgggcgggg cggggccgcc tcgggccggg
     3901 gagggctcgg gggaggggcg cggcggcccc cggagcgccg gcggctgtcg aggcgcggcg
     3961 agccgcagcc attgcctttt atggtaatcg tgcgagaggg cgcagggact tcctttgtcc
     4021 caaatctgtg cggagccgaa atctgggagg cgccgccgca ccccctctag cgggcgcggg
     4081 gcgaagcggt gcggcgccgg caggaaggaa atgggcgggg agggccttcg tgcgtcgccg
     4141 cgccgccgtc cccttctccc tctccagcct cggggctgtc cgcgggggga cggctgcctt
     4201 cgggggggac ggggcagggc ggggttcggc ttctggcgtg tgaccggcgg ctctagagcc
     4261 tctgctaacc atgttcatgc cttcttcttt ttcctacagc tcctgggcaa cgtgctggtt
     4321 attgtgctgt ctcatcattt tggcaaagaa ttaattcaag cttgggccac cggctagcat
     4381 gacttctgag aaaggtcctt caaccggtga ccccactctg aggagaagaa tcgaaccctg
     4441 ggagtttgac gtcttctatg accccagaga acttcgtaaa gaggcctgtc tgctctacga
     4501 aatcaagtgg ggcatgagcc ggaaaatctg gcgaagctca ggcaaaaaca ccaccaatca
     4561 cgtggaagtt aattttataa aaaaatttac gtcagaaaga gattttcacc catccatcag
     4621 ctgctccatc acctggttct tgtcctggag tccctgctgg gaatgctccc aggctattag
     4681 agagtttctg agtcggcacc ctggtgtgac tctagtgatc tacgtagctc ggcttttttg
     4741 gcacatggat caacaaaatc ggcaaggtct cagggacctt gttaacagtg gagtaactat
     4801 tcagattatg agagcatcag agtattatca ctgctggagg aattttgtca actacccacc
     4861 tggggatgaa gctcactggc cacaataccc acctctgtgg atgatgttgt acgcactgga
     4921 gctgcactgc ataattctaa gtcttccacc ctgtttaaag atttcaagaa gatggcaaaa
     4981 tcatcttaca tttttcagac ttcatcttca aaactgccat taccaaacga ttccgccaca
     5041 catcctttta gctacagggc tgatacatcc ttctgtggct tggagatgag atcttgtaca
     5101 agtccggact cagatcctag cccggggcgg ccccgccgac cccgcacccg cccccgaggc
     5161 ccaccgactc tagaggatca taatcagcca taccacattt gtagaggttt tacttgcttt
     5221 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
     5281 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     5341 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     5401 ttatcatgtc tggatctgga ctagcccaga tcctcgaggt gagccccacg ttctgcttca
     5461 ctctccccat ctcccccccc tccccacccc caattttgta tttatttatt ttttaattat
     5521 tttgtgcagc gatgggggcg gggggggggg gggcgcgcgc caggcggggc ggggcggggc
     5581 gaggggcggg gcggggcgag gcggagaggt gcggcggcag ccaatcagag cggcgcgctc
     5641 cgaaagtttc cttttatggc gaggcggcgg cggcggcggc cctataaaaa gcgaagcgcg
     5701 cggcgggcgg gagtcgctgc gttgccttcg ccccgtgccc cgctccgcgc cgcctcgcgc
     5761 cgcccgcccc ggctctgact gaccgcgtta ctcccacagg tgagcgggcg ggacggccct
     5821 tctcctccgg gctgtaatta gcgcttggtt taatgacggc tcgtttcttt tctgtggctg
     5881 cgtgaaagcc ttaaagggct ccgggagggc cctttgtgcg ggggggagcg gctcgggggg
     5941 tgcgtgcgtg tgtgtgtgcg tggggagcgc cgcgtgcggc ccgcgctgcc cggcggctgt
     6001 gagcgctgcg ggcgcggcgc ggggctttgt gcgctccgcg tgtgcgcgag gggagcgcgg
     6061 ccgggggcgg tgccccgcgg tgcggggggg ctgcgagggg aacaaaggct gcgtgcgggg
     6121 tgtgtgcgtg ggggggtgag cagggggtgt gggcgcggcg gtcgggctgt aacccccccc
     6181 tgcacccccc tccccgagtt gctgagcacg gcccggcttc gggtgcgggg ctccgtgcgg
     6241 ggcgtggcgc ggggctcgcc gtgccgggcg gggggtggcg gcaggtgggg gtgccgggcg
     6301 gggcggggcc gcctcgggcc ggggagggct cgggggaggg gcgcggcggc ccccggagcg
     6361 ccggcggctg tcgaggcgcg gcgagccgca gccattgcct tttatggtaa tcgtgcgaga
     6421 gggcgcaggg acttcctttg tcccaaatct gtgcggagcc gaaatctggg aggcgccgcc
     6481 gcaccccctc tagcgggcgc ggggcgaagc ggtgcggcgc cggcaggaag gaaatgggcg
     6541 gggagggcct tcgtgcgtcg ccgcgccgcc gtccccttct ccctctccag cctcggggct
     6601 gtccgcgggg ggacggctgc cttcgggggg gacggggcag ggcggggttc ggcttctggc
     6661 gtgtgaccgg cggctctaga gcctctgcta accatgttca tgccttcttc tttttcctac
     6721 agctcctggg caacgtgctg gttattgtgc tgtctcatca ttttggcaaa gaattaattc
     6781 aagcttgcat gcctgcaggt cggccgccac gaccggtgcc gccaccatcc cctgacccac
     6841 gcccctgacc cctcacaagg agacgacctt ccatgaccga gtacaagccc acggtgcgcc
     6901 tcgccacccg cgacgacgtc ccccgggccg tacgcaccct cgccgccgcg ttcgccgact
     6961 accccgccac gcgccacacc gtcgacccgg accgccacat cgagcgggtc accgagctgc
     7021 aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg
     7081 gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg
     7141 agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg
     7201 aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg
     7261 tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg
     7321 cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct
     7381 tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca
     7441 cctggtgcat gacccgcaag cccggtgcct gacgcccgcc ccacgacccg cagcgcccga
     7501 ccgaaaggag cgcacgaccc catggctccg accgaagccg acccgggcgg ccccgccgac
     7561 cccgcacccg cccccgaggc ccaccgactc tagaggatca taatcagcca taccacattt
     7621 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa
     7681 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
     7741 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     7801 tccaaactca tcaatgtatc ttatcatgtc tggatctata acttcgtata gcatacatta
     7861 tacgaacggt aggggatcc
//
