LOCUS       sequence-213790-        7232 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of human catalitic dead Exonuclease 1 (EXO1) 
            mCherry-tagged in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7232)
  AUTHORS   Sertic S, Mollica A, Campus I, Roma S, Tumini E, Aguilera A, 
            Muzi-Falconi M
  TITLE     Coordinated Activity of Y Family TLS Polymerases and EXO1 Protects 
            Non-S Phase Cells from UV-Induced Cytotoxic Lesions.
  JOURNAL   Mol Cell. 2018 Apr 5;70(1):34-47.e4. doi: 
            10.1016/j.molcel.2018.02.017. Epub 2018 Mar 15.
  PUBMED    29551515
REFERENCE   2  (bases 1 to 7232)
  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"; doi: 
            "10.1016/j.molcel.2018.02.017"; journalName: "Mol Cell"; date: 
            "2018-04-5- 5"; volume: "70"; issue: "1"; pages: "34-47.e4"
COMMENT     Sequence Label: pmCherry-N1-EXO1b-D173A
FEATURES             Location/Qualifiers
     source          1..7232
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        284..487
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     438..458
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3100..3109
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3106..3816
                     /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(3251..3269)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3519..3538
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3939..4060
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3976..3995)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4030..4049
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4067..4522)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4204..4225)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4416..4435
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4549..4653
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4655..5012
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4655..4846
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4678..4698)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4863..4998
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5047..5841
                     /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             5047..5841
                     /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(5101..5120)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5711..5730
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6029..6048)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6073..6120
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6449..7037
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6938..6957
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        join(7212..7232,1..283)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
       61 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
      121 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
      181 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
      241 tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca
      301 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
      361 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
      421 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
      481 cagagctggt ttagtgaacc gtcagatccg ctagcgctac cggactcaga tctcgagctc
      541 aagcttgctt cgatggggat acagggattg ctacaattta tcaaagaagc ttcagaaccc
      601 atccatgtga ggaagtataa agggcaggta gtagctgtgg atacatattg ctggcttcac
      661 aaaggagcta ttgcttgtgc tgaaaaacta gccaaaggtg aacctactga taggtatgta
      721 ggattttgta tgaaatttgt aaatatgtta ctatctcatg ggatcaagcc tattctcgta
      781 tttgatggat gtactttacc ttctaaaaag gaagtagaga gatctagaag agaaagacga
      841 caagccaatc ttcttaaggg aaagcaactt cttcgtgagg ggaaagtctc ggaagctcga
      901 gagtgtttca cccggtctat caatatcaca catgccatgg cccacaaagt aattaaagct
      961 gcccggtctc agggggtaga ttgcctcgtg gctccctatg aagctgatgc gcagttggcc
     1021 tatcttaaca aagcgggaat tgtgcaagcc ataattacag aggactcggc tctcctagct
     1081 tttggctgta aaaaggtaat tttaaagatg gaccagtttg gaaatggact tgaaattgat
     1141 caagctcggc taggaatgtg cagacagctt ggggatgtat tcacggaaga gaagtttcgt
     1201 tacatgtgta ttctttcagg ttgtgactac ctgtcatcac tgcgtgggat tggattagca
     1261 aaggcatgca aagtcctaag actagccaat aatccagata tagtaaaggt tatcaagaaa
     1321 attggacatt atctcaagat gaatatcacg gtaccagagg attacatcaa cgggtttatt
     1381 cgggccaaca ataccttcct ctatcagcta gtttttgatc ccatcaaaag gaaacttatt
     1441 cctctgaacg cctatgaaga tgatgttgat cctgaaacac taagctacgc tgggcaatat
     1501 gttgatgatt ccatagctct tcaaatagca cttggaaata aagatataaa tacttttgaa
     1561 cagatcgatg actacaatcc agacactgct atgcctgccc attcaagaag tcgtagttgg
     1621 gatgacaaaa catgtcaaaa gtcagctaat gttagcagca tttggcatag gaattactct
     1681 cccagaccag agtcgggtac tgtttcagat gccccacaat tgaaggaaaa tccaagtact
     1741 gtgggagtgg aacgagtgat tagtactaaa gggttaaatc tcccaaggaa atcatccatt
     1801 gtgaaaagac caagaagtgc agagctgtca gaagatgacc tgttgagtca gtattctctt
     1861 tcatttacga agaagaccaa gaaaaatagc tctgaaggca ataaatcatt gagcttttct
     1921 gaagtgtttg tgcctgacct ggtaaatgga cctactaaca aaaagagtgt aagcactcca
     1981 cctaggacga gaaataaatt tgcaacattt ttacaaagga aaaatgaaga aagtggtgca
     2041 gttgtggttc cagggaccag aagcaggttt ttttgcagtt cagattctac tgactgtgta
     2101 tcaaacaaag tgagcatcca gcctctggat gaaactgctg tcacagataa agagaacaat
     2161 ctgcatgaat cagagtatgg agaccaagaa ggcaagagac tggttgacac agatgtagca
     2221 cgtaattcaa gtgatgacat tccgaataat catattccag gtgatcatat tccagacaag
     2281 gcaacagtgt ttacagatga agagtcctac tcttttaaga gcagcaaatt tacaaggacc
     2341 atttcaccac ccactttggg aacactaaga agttgtttta gttggtctgg aggtcttgga
     2401 gatttttcaa gaacgccgag cccctctcca agcacagcat tgcagcagtt ccgaagaaag
     2461 agcgattccc ccacctcttt gcctgagaat aatatgtctg atgtgtcgca gttaaagagc
     2521 gaggagtcca gtgacgatga gtctcatccc ttacgagaag gggcatgttc ttcacagtcc
     2581 caggaaagtg gagaattctc actgcagagt tcaaatgcat caaagctttc tcagtgctct
     2641 agtaaggact ctgattcaga ggaatctgat tgcaatatta agttacttga cagtcaaagt
     2701 gaccagacct ccaagctatg tttatctcat ttctcaaaaa aagacacacc tctaaggaac
     2761 aaggttcctg ggctatataa gtccagttct gcagactctc tttctacaac caagatcaaa
     2821 cctctaggac ctgccagagc cagtgggctg agcaagaagc cggcaagcat ccagaagaga
     2881 aagcatcata atgccgagaa caagccgggg ttacagatca aactcaatga gctctggaaa
     2941 aactttggat ttaaaaaaga ttctgaaaag cttcctcctt gtaagaaacc cctgtcccca
     3001 gtcagagata acatccaact aactccagaa gcggaagagg atatatttaa caaacctgaa
     3061 tgtggccgtg ttcaaagagc aatattccag cgggatcccg ccaccatggt gagcaagggc
     3121 gaggaggata acatggccat catcaaggag ttcatgcgct tcaaggtgca catggagggc
     3181 tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc
     3241 acccagaccg ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc
     3301 ctgtcccctc agttcatgta cggctccaag gcctacgtga agcaccccgc cgacatcccc
     3361 gactacttga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag
     3421 gacggcggcg tggtgaccgt gacccaggac tcctccctgc aggacggcga gttcatctac
     3481 aaggtgaagc tgcgcggcac caacttcccc tccgacggcc ccgtaatgca gaagaagact
     3541 atgggctggg aggcctcctc cgagcggatg taccccgagg acggcgccct gaagggcgag
     3601 atcaagcaga ggctgaagct gaaggacggc ggccactacg acgctgaggt caagaccacc
     3661 tacaaggcca agaagcccgt gcagctgccc ggcgcctaca acgtcaacat caagttggac
     3721 atcacctccc acaacgagga ctacaccatc gtggaacagt acgaacgcgc cgagggccgc
     3781 cactccaccg gcggcatgga cgagctgtac aagtaaagcg gccgcgactc tagatcataa
     3841 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
     3901 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
     3961 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     4021 attctagttg tggtttgtcc aaactcatca atgtatctta aggcgtaaat tgtaagcgtt
     4081 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     4141 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     4201 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     4261 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     4321 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     4381 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     4441 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     4501 aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     4561 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     4621 taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga
     4681 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     4741 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     4801 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     4861 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     4921 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     4981 gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg
     5041 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     5101 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     5161 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     5221 gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     5281 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     5341 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     5401 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     5461 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     5521 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg
     5581 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     5641 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     5701 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     5761 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     5821 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     5881 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     5941 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     6001 tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa
     6061 cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc
     6121 ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt
     6181 ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag
     6241 gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat
     6301 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     6361 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     6421 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     6481 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     6541 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     6601 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     6661 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     6721 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     6781 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     6841 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     6901 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     6961 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     7021 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     7081 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     7141 ccgccatgca ttagttatta atagtaatca attacggggt cattagttca tagcccatat
     7201 atggagttcc gcgttacata acttacggta aa
//
