LOCUS       sequence-335363-        4627 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses rat APOBEC1 L173A/G227A in mammalian cells. Please note 
            that this does not express GFP..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4627)
  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 4627)
  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: pEGFP ratAPOBEC1 L173A/G227A
FEATURES             Location/Qualifiers
     source          1..4627
                     /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             537..1226
                     /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
                     EAHWPRYPHAWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHI
                     LWATALK"
     polyA_signal    1355..1476
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1392..1411)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1446..1465
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1483..1938)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1620..1641)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1832..1851
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1965..2069
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2071..2428
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2071..2262
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2094..2114)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2279..2414
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2463..3257
                     /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             2463..3257
                     /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(2517..2536)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3127..3146
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3445..3464)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3489..3536
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3865..4453
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4354..4373
                     /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 gctagcatga
      541 gttccgagac aggccctgta gctgttgatc ccactctgag gagaagaatt gagccccacg
      601 agtttgaagt cttctttgac ccccgggaac ttcggaaaga gacctgtctg ctgtatgaga
      661 tcaactgggg aggaaggcac agcatctggc gacacacgag ccaaaacacc aacaaacacg
      721 ttgaagtcaa tttcatagaa aaatttacta cagaaagata cttttgtcca aacaccagat
      781 gctccattac ctggttcctg tcctggagtc cctgtgggga gtgctccagg gccattacag
      841 aatttttgag ccgatacccc catgtaactc tgtttattta tatagcacgg ctttatcacc
      901 acgcagatcc tcgaaatcgg caaggactca gggaccttat tagcagcggt gttactatcc
      961 agatcatgac ggagcaagag tctggctact gctggaggaa ttttgtcaac tactcccctt
     1021 cgaatgaagc tcattggcca aggtaccccc atgcgtgggt gaggctgtac gtactggaac
     1081 tctactgcat cattttagga cttccaccct gtttaaatat tttaagaaga aaacaacctc
     1141 aactcacgtt tttcacgatt gctcttcaaa gctgccatta ccaaaggcta ccaccccaca
     1201 tcctgtgggc cacagcgttg aaatgagatc caccggatct agataactga tcataatcag
     1261 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa
     1321 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
     1381 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     1441 tagttgtggt ttgtccaaac tcatcaatgt atcttaacgc gtaaattgta agcgttaata
     1501 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     1561 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     1621 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     1681 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     1741 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     1801 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     1861 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     1921 cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     1981 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     2041 tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt
     2101 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     2161 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     2221 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     2281 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     2341 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     2401 cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc
     2461 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     2521 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     2581 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     2641 tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     2701 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     2761 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     2821 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     2881 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     2941 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg
     3001 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     3061 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     3121 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     3181 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     3241 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     3301 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     3361 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     3421 cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg
     3481 cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa
     3541 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg
     3601 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc
     3661 agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat
     3721 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     3781 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     3841 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     3901 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     3961 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     4021 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     4081 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     4141 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     4201 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     4261 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     4321 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     4381 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     4441 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     4501 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     4561 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
     4621 agttccg
//
