LOCUS       sequence-325012-        6209 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 6209)
  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 6209)
  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: Nsp4-mCherry
FEATURES             Location/Qualifiers
     source          1..6209
                     /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"
     misc_feature    1740..1792
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      1794..1803
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1800..2510
                     /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(1945..1963)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2213..2232
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2633..2754
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2670..2689)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2724..2743
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2761..3216)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2898..2919)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3110..3129
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3243..3347
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3349..3706
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3349..3540
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3372..3392)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3557..3692
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3741..4535
                     /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             3741..4535
                     /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(3795..3814)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4405..4424
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4723..4742)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4767..4814
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5143..5731
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5632..5651
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        5906..6209
                     /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 gccaccatga
      241 agattgtaaa caattggctt aaacaactta ttaaagtaac cctggtattc ttgtttgtcg
      301 ccgcaatctt ttacttgata acaccggtcc acgtgatgtc taaacacaca gactttagta
      361 gtgagataat cggatataaa gctatcgatg gcggcgttac ccgcgacatc gcttcaaccg
      421 acacatgttt cgctaataag cacgccgatt ttgacacttg gtttagtcaa agaggaggaa
      481 gctacacaaa cgacaaggct tgtccgctta tcgcggctgt aataacaagg gaagttgggt
      541 ttgtcgtgcc tggtttgcca ggtactatac tcagaaccac aaacggtgac tttctgcact
      601 tcttgccaag ggtttttagc gcggtaggca atatctgcta tacgccaagc aagctgattg
      661 aatacaccga ctttgcgacg agcgcatgtg tcctcgctgc agagtgtacg atatttaagg
      721 atgcgagcgg aaagcccgta ccttactgct atgatactaa tgtccttgag ggatcagttg
      781 cgtatgagag tcttaggccg gatacacgat atgtgctcat ggatggttcc ataatccagt
      841 tcccaaatac ctacctcgaa gggtccgttc gggtcgtcac cacatttgat agcgaatatt
      901 gcagacacgg aacatgcgaa cggagtgaag caggggtctg cgtctccaca agtgggcggt
      961 gggtcctcaa taacgattat taccggagcc tgcccggcgt gttttgcggg gtcgatgcgg
     1021 ttaacttgtt gacgaatatg tttacgcctc tcatacaacc cattggcgca ctggatatat
     1081 ctgcaagtat agtcgctgga ggcattgtgg caatcgtggt tacgtgtttg gcgtattatt
     1141 ttatgaggtt ccgccgggct ttcggggaat attcccacgt agtcgccttc aacacactct
     1201 tgtttctcat gtctttcact gtactgtgtt tgactcctgt ctactctttc ttgccgggtg
     1261 tatactcagt tatttacttg tacctcacgt tttatttgac aaacgatgtt tcatttctcg
     1321 cgcatatcca atggatggtg atgtttacgc cattggtgcc tttctggatc acgatcgctt
     1381 atatcatttg catcagcacc aaacacttct attggttctt tagtaactac ttgaagcgca
     1441 gggttgtttt taacggtgtc agcttctcaa cgtttgaaga ggcagcactg tgtactttcc
     1501 ttctgaataa ggaaatgtat ctcaagctgc ggtccgatgt cctgcttccg cttacccagt
     1561 ataatcgcta tctcgccctg tataataagt acaagtactt ctcaggcgcg atggatacaa
     1621 ccagttatag agaggctgcc tgctgccatc ttgccaaagc tttgaatgac ttcagcaata
     1681 gtggctcaga cgttctctac caaccaccgc aaacttccat aacctcagcc gtactccagc
     1741 tcgagctcaa gcttcgaatt ctgcagtcga cggtaccgcg ggcccgggat cccgccacca
     1801 tggtgagcaa gggcgaggag gataacatgg ccatcatcaa ggagttcatg cgcttcaagg
     1861 tgcacatgga gggctccgtg aacggccacg agttcgagat cgagggcgag ggcgagggcc
     1921 gcccctacga gggcacccag accgccaagc tgaaggtgac caagggtggc cccctgccct
     1981 tcgcctggga catcctgtcc cctcagttca tgtacggctc caaggcctac gtgaagcacc
     2041 ccgccgacat ccccgactac ttgaagctgt ccttccccga gggcttcaag tgggagcgcg
     2101 tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg
     2161 gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt cccctccgac ggccccgtaa
     2221 tgcagaagaa gaccatgggc tgggaggcct cctccgagcg gatgtacccc gaggacggcg
     2281 ccctgaaggg cgagatcaag cagaggctga agctgaagga cggcggccac tacgacgctg
     2341 aggtcaagac cacctacaag gccaagaagc ccgtgcagct gcccggcgcc tacaacgtca
     2401 acatcaagtt ggacatcacc tcccacaacg aggactacac catcgtggaa cagtacgaac
     2461 gcgccgaggg ccgccactcc accggcggca tggacgagct gtacaagtaa agcggccgcg
     2521 actctagatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct
     2581 cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt
     2641 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     2701 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaaggcgt
     2761 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     2821 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
     2881 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     2941 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
     3001 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     3061 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     3121 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     3181 cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg
     3241 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     3301 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa
     3361 gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     3421 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     3481 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     3541 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     3601 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     3661 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga
     3721 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     3781 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     3841 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     3901 ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg
     3961 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     4021 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     4081 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     4141 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     4201 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     4261 tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     4321 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     4381 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     4441 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     4501 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     4561 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     4621 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     4681 tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac
     4741 aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt
     4801 tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc
     4861 accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc
     4921 aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc
     4981 ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     5041 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     5101 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     5161 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     5221 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     5281 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     5341 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     5401 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     5461 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     5521 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     5581 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     5641 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     5701 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     5761 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     5821 ggataaccgt attaccgcca tgcattagtt attaatagta atcaattacg gggtcattag
     5881 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
     5941 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
     6001 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
     6061 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
     6121 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     6181 tctacgtatt agtcatcgct attaccatg
//
