LOCUS       sequence-254328-        5814 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid bearing the editing target of APOBEC1 fused between the 
            mCherry and the Tag-BFP one. In presence of APOBEC1, the CAA codon 
            in ApoB is edited to a Stop codon (UAA), leading to loss of Tag-BFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5814)
  AUTHORS   Chieca M, Torrini S, Conticello SG
  TITLE     Live-Cell Quantification of APOBEC1-Mediated RNA Editing: A 
            Comparison of RNA Editing Assays.
  JOURNAL   Methods Mol Biol. 2021;2181:69-81. doi: 10.1007/978-1-0716-0787-9_5.
  PUBMED    32729075
REFERENCE   2  (bases 1 to 5814)
  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: "Methods Mol
            Biol."; date: "2021"; volume: "2181:69-81. doi"; pages: " 
            10.1007/978-1-0716-0787-9_5"
COMMENT     Sequence Label: pEGFP-N1-mCherry-ApoB-TagBFP
FEATURES             Location/Qualifiers
     source          1..5814
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             546..1253
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(691..709)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     959..978
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1740..2441
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     polyA_signal    2563..2684
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2600..2619)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2654..2673
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2691..3146)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2828..2849)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3040..3059
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3173..3277
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        3279..3470
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3302..3322)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             3650..4444
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3650..4444
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3704..3723)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4314..4333
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4632..4651)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4676..4723
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5052..5640
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5541..5560
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcggat
      541 tcaccatggt gagcaagggc gaggaggata acatggccat catcaaggag ttcatgcgct
      601 tcaaggtgca catggagggc tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg
      661 agggccgccc ctacgagggc acccagaccg ccaagctgaa ggtgaccaag ggtggccccc
      721 tgcccttcgc ctgggacatc ctgtcccctc agttcatgta cggctccaag gcctacgtga
      781 agcaccccgc cgacatcccc gactacttga agctgtcctt ccccgagggc ttcaagtggg
      841 agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt gacccaggac tcctccctgc
      901 aggacggcga gttcatctac aaggtgaagc tgcgcggcac caacttcccc tccgacggcc
      961 ccgtaatgca gaagaagacc atgggctggg aggcctcctc cgagcggatg taccccgagg
     1021 acggcgccct gaagggcgag atcaagcaga ggctgaagct gaaggacggc ggccactacg
     1081 acgctgaggt caagaccacc tacaaggcca agaagcccgt gcagctgccc ggcgcctaca
     1141 acgtcaacat caagttggac atcacctccc acaacgagga ctacaccatc gtggaacagt
     1201 acgaacgcgc cgagggccgc cactccaccg gcggcatgga cgagctgtac aagaagctta
     1261 ccatggccaa ggagaaactg actgctctca caaaaaagta tagaattaca gaaaatgata
     1321 tacaaattgc attagatgat gccaaaatca actttaatga aaaactatct caactgcaga
     1381 catatatgat acaatttgat cagtatatta aagatagtta tgatttacat gatttgaaaa
     1441 tagctattgc taatattatt gatgaaatca ttgaaaaatt aaaaagtctt gatgagcact
     1501 atcatatccg tgtaaattta gtaaaaacaa tccatgatct acatttgttt attgaaaata
     1561 ttgattttaa caaaagtgga agtagtactg catcctggat tcaaaatgtg gatactaagt
     1621 accaaatcag aatccagata caagaaaaac tgcagcagct taagagacac atacagaata
     1681 tagacatcca gcacctagct ggaattctgc agtcgacggt accgcgggcc cgggatccaa
     1741 tgagcgagct gattaaggag aacatgcaca tgaagctgta catggagggc accgtggaca
     1801 accatcactt caagtgcaca tccgagggcg aaggcaagcc ctacgagggc acccagacca
     1861 tgagaatcaa ggtggtcgag ggcggccctc tccccttcgc cttcgacatc ctggctacta
     1921 gcttcctcta cggcagcaag accttcatca accacaccca gggcatcccc gacttcttca
     1981 agcagtcctt ccctgagggc ttcacatggg agagagtcac cacatacgaa gacgggggcg
     2041 tgctgaccgc tacccaggac accagcctcc aggacggctg cctcatctac aacgtcaaga
     2101 tcagaggggt gaacttcaca tccaacggcc ctgtgatgca gaagaaaaca ctcggctggg
     2161 aggccttcac cgagacgctg taccccgctg acggcggcct ggaaggcaga aacgacatgg
     2221 ccctgaagct cgtgggcggg agccatctga tcgcaaacat caagaccaca tatagatcca
     2281 agaaacccgc taagaacctc aagatgcctg gcgtctacta tgtggactac agactggaaa
     2341 gaatcaagga ggccaacaac gagacctacg tcgagcagca cgaggtggca gtggccagat
     2401 actgcgacct ccctagcaaa ctggggcaca aacttaatta agcggccgcg actctagatc
     2461 ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc
     2521 cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct
     2581 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     2641 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaaggcgt aaattgtaag
     2701 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
     2761 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag
     2821 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
     2881 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt
     2941 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
     3001 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
     3061 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
     3121 gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac
     3181 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     3241 ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa gaaccagctg
     3301 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3361 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     3421 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     3481 ccgcccatcc cgcccctaac tccgccccat ggctgactaa ttttttttat ttatgcagag
     3541 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     3601 ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca
     3661 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     3721 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     3781 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc
     3841 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     3901 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     3961 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     4021 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     4081 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     4141 gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct
     4201 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     4261 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     4321 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     4381 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     4441 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     4501 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     4561 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg
     4621 gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca
     4681 ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg
     4741 tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc
     4801 gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc
     4861 aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat
     4921 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     4981 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     5041 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     5101 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     5161 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     5221 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     5281 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     5341 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     5401 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     5461 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     5521 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     5581 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     5641 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     5701 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat gcattagtta
     5761 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccg
//
