LOCUS       sequence-218644-        5410 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  plasmid for the expression of a rat APOBEC1-EGFP C-terminal fusion 
            protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5410)
  AUTHORS   Severi F, Conticello SG
  TITLE     Flow-cytometric visualization of C>U mRNA editing reveals the 
            dynamics of the process in live cells.
  JOURNAL   RNA Biol. 2015;12(4):389-97. doi: 10.1080/15476286.2015.1026033.
  PUBMED    25806564
REFERENCE   2  (bases 1 to 5410)
  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: "RNA Biol.";
            date: "2015"; volume: "12(4):389-97. doi"; pages: " 
            10.1080/15476286.2015.1026033"
COMMENT     Sequence Label: rat APOBEC1-EGFP
FEATURES             Location/Qualifiers
     source          1..5410
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             254..937
                     /codon_start=1
                     /product="cytidine deaminase (C to U editing enzyme) from 
                     rat"
                     /label=APOBEC-1
                     /note="can use ssDNA as a substrate (Komor et al., 2016)"
                     /translation="MSSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWG
                     GRHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFL
                     SRYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSN
                     EAHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHI
                     LWATGL"
     regulatory      986..995
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(992..994,995..997,998..1711)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(992..994,995..997,998..1708)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1037..1058)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1298..1317)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1645..1666
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1834..1955
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1871..1890)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1925..1944
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1962..2417)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2099..2120)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2311..2330
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2444..2548
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2550..2907
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2550..2741
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2573..2593)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2758..2893
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2942..3736
                     /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             2942..3736
                     /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(2996..3015)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3606..3625
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3924..3943)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3968..4015
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4344..4932
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4833..4852
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        5107..5410
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta gcgctaccgg
      241 actcagatct accatgagtt ccgagacagg ccctgtagct gttgatccca ctctgaggag
      301 aagaattgag ccccacgagt ttgaagtctt ctttgacccc cgggaacttc ggaaagagac
      361 ctgtctgctg tatgagatca actggggagg aaggcacagc atctggcgac acacgagcca
      421 aaacaccaac aaacacgttg aagtcaattt catagaaaaa tttactacag aaagatactt
      481 ttgtccaaac accagatgct ccattacctg gttcctgtcc tggagtccct gtggggagtg
      541 ctccagggcc attacagaat ttttgagccg atacccccat gtaactctgt ttatttatat
      601 agcacggctt tatcaccacg cagatcctcg aaatcggcaa ggactcaggg accttattag
      661 cagcggtgtt actatccaga tcatgacgga gcaagagtct ggctactgct ggaggaattt
      721 tgtcaactac tccccttcga atgaagctca ttggccaagg tacccccatc tgtgggtgag
      781 gctgtacgta ctggaactct actgcatcat tttaggactt ccaccctgtt taaatatttt
      841 aagaagaaaa caacctcaac tcacgttttt cacgattgct cttcaaagct gccattacca
      901 aaggctacca ccccacatcc tgtgggccac agggttgaat agaattctgc agtcgacggt
      961 accgcgggcc cgggatccac cggtcgccac catggtgagc aagggcgagg agctgttcac
     1021 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
     1081 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac
     1141 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca
     1201 gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
     1261 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
     1321 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     1381 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
     1441 cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca
     1501 caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
     1561 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa
     1621 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
     1681 cactctcggc atggacgagc tgtacaagta aagcggccgc gactctagat cataatcagc
     1741 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac
     1801 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt
     1861 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     1921 agttgtggtt tgtccaaact catcaatgta tcttaaggcg taaattgtaa gcgttaatat
     1981 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     2041 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     2101 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     2161 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     2221 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     2281 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     2341 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     2401 gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     2461 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     2521 gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg
     2581 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     2641 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     2701 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     2761 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     2821 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     2881 ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg
     2941 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     3001 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     3061 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     3121 gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     3181 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     3241 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     3301 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     3361 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     3421 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga
     3481 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     3541 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     3601 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     3661 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     3721 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     3781 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     3841 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     3901 gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc
     3961 gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa
     4021 cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc
     4081 caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca
     4141 gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata
     4201 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     4261 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     4321 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     4381 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     4441 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     4501 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     4561 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     4621 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     4681 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     4741 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     4801 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     4861 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     4921 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     4981 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     5041 atgcattagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
     5101 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
     5161 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
     5221 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
     5281 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
     5341 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
     5401 tattaccatg
//
