LOCUS       sequence-325015-        6290 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression and localization.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6290)
  AUTHORS   Miserey-Lenkei S, Trajkovic K, D'Ambrosio JM, Patel AJ, Copic A, 
            Mathur P, Schauer K, Goud B, Albanese V, Gautier R, Subra M, Kovacs 
            D, Barelli H, Antonny B
  TITLE     A comprehensive library of fluorescent constructs of SARS-CoV-2 
            proteins and their initial characterisation in different cell types.
  JOURNAL   Biol Cell. 2021 Jul;113(7):311-328. doi: 10.1111/boc.202000158. Epub
            2021 May 10.
  PUBMED    33666950
REFERENCE   2  (bases 1 to 6290)
  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: 
            "10.1111/boc.202000158"; journalName: "Biol Cell"; date: "2021-07"; 
            volume: "113"; issue: "7"; pages: "311-328"
COMMENT     Sequence Label: Nsp14-mCherry
FEATURES             Location/Qualifiers
     source          1..6290
                     /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"
     regulatory      535..544
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    2125..2177
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      2179..2188
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2185..2895
                     /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(2330..2348)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2598..2617
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3018..3139
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3055..3074)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3109..3128
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3146..3601)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3283..3304)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3495..3514
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3628..3732
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3734..4091
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3734..3925
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3757..3777)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3942..4077
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4126..4920
                     /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             4126..4920
                     /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(4180..4199)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4790..4809
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5108..5127)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5152..5199
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5528..6116
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6017..6036
                     /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 gctagccacc
      541 atggctgaaa atgtaacggg cttgtttaaa gactgtagta aagtgatcac aggactccac
      601 cccacacaag cacctaccca cctttccgta gatacgaagt tcaaaacgga aggattgtgt
      661 gtggatatac cagggatacc aaaggatatg acgtaccgaa ggctgatttc catgatgggt
      721 tttaagatga attaccaagt taatggctac cccaacatgt tcatcaccag ggaggaggca
      781 attagacacg taagagcctg gataggcttc gatgttgaag gttgccatgc aacccgggaa
      841 gccgtaggca caaaccttcc gttgcagctt ggcttttcca cgggcgtcaa cctcgttgcc
      901 gtaccgactg gctatgttga cacgccgaac aacaccgatt tctctcgcgt aagtgctaag
      961 cctcctccgg gagatcaatt caagcatctt atacctctca tgtacaaggg actgccgtgg
     1021 aatgtagttc gcattaagat cgtgcagatg ctctccgata ctttgaaaaa tctgagcgac
     1081 agggtggtct tcgttctgtg ggcacatggc ttcgaactta cttctatgaa atactttgta
     1141 aaaatcgggc ccgagcggac atgttgtttg tgcgaccgac gagctacttg ttttagcaca
     1201 gcatctgaca cctacgcatg ctggcaccac agtataggct tcgattacgt ctacaatccc
     1261 tttatgatag acgtacaaca atggggcttt acgggtaact tgcagagtaa tcacgatctt
     1321 tactgccaag ttcatgggaa cgcacacgtg gcctcctgcg atgcgataat gacgaggtgc
     1381 ttggccgtgc acgagtgctt tgttaagcgg gtcgattgga ctatagagta tcccataatc
     1441 ggtgacgaac ttaaaattaa tgctgcatgc aggaaagtgc agcacatggt agtaaaagca
     1501 gcgctcctcg cggataagtt tcctgtgctc cacgatatcg gcaaccctaa ggccattaaa
     1561 tgcgtccccc aagctgatgt agagtggaaa ttctatgacg ctcagccatg tagtgacaaa
     1621 gcatacaaga ttgaagaatt gttttattca tatgcgacac actccgataa gttcactgac
     1681 ggagtttgtc tcttttggaa ctgtaatgtt gataggtatc cggcgaatag catagtgtgt
     1741 cgatttgaca cgcgcgttct ctctaatttg aacctccctg gctgcgacgg ggggtctttg
     1801 tacgtaaaca agcatgcatt tcatacccca gcttttgaca aatccgcttt cgtaaatctg
     1861 aaacaacttc ccttcttcta ttattcagat agcccttgcg aatctcacgg taagcaagtg
     1921 gttagtgaca ttgactatgt accacttaaa agtgcaacgt gcattacacg atgcaatttg
     1981 gggggcgcgg tttgccgaca ccatgccaac gaatacagat tgtacctgga tgcatataac
     2041 atgatgataa gtgcagggtt tagtctttgg gtgtataagc agttcgatac gtataatctc
     2101 tggaacacat ttacccgatt gcagctcgag ctcaagcttc gaattctgca gtcgacggta
     2161 ccgcgggccc gggatcccgc caccatggtg agcaagggcg aggaggataa catggccatc
     2221 atcaaggagt tcatgcgctt caaggtgcac atggagggct ccgtgaacgg ccacgagttc
     2281 gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag
     2341 gtgaccaagg gtggccccct gcccttcgcc tgggacatcc tgtcccctca gttcatgtac
     2401 ggctccaagg cctacgtgaa gcaccccgcc gacatccccg actacttgaa gctgtccttc
     2461 cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg
     2521 acccaggact cctccctgca ggacggcgag ttcatctaca aggtgaagct gcgcggcacc
     2581 aacttcccct ccgacggccc cgtaatgcag aagaagacca tgggctggga ggcctcctcc
     2641 gagcggatgt accccgagga cggcgccctg aagggcgaga tcaagcagag gctgaagctg
     2701 aaggacggcg gccactacga cgctgaggtc aagaccacct acaaggccaa gaagcccgtg
     2761 cagctgcccg gcgcctacaa cgtcaacatc aagttggaca tcacctccca caacgaggac
     2821 tacaccatcg tggaacagta cgaacgcgcc gagggccgcc actccaccgg cggcatggac
     2881 gagctgtaca agtaaagcgg ccgcgactct agatcataat cagccatacc acatttgtag
     2941 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
     3001 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     3061 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     3121 aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg
     3181 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     3241 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     3301 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     3361 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     3421 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     3481 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     3541 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     3601 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     3661 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     3721 aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg
     3781 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3841 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3901 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     3961 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     4021 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     4081 gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     4141 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     4201 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     4261 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg
     4321 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     4381 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     4441 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     4501 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     4561 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     4621 gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc
     4681 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     4741 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     4801 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     4861 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     4921 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     4981 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     5041 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga
     5101 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa
     5161 aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc
     5221 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt
     5281 cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg
     5341 tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag attgatttaa
     5401 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     5461 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     5521 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     5581 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     5641 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     5701 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     5761 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     5821 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     5881 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     5941 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     6001 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     6061 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     6121 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     6181 ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat tagttattaa
     6241 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
//
