LOCUS       sequence-324467-        5531 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 5531)
  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 5531)
  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: Orf3a-mCherry
FEATURES             Location/Qualifiers
     source          1..5531
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        128..319
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     276..296
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(321..425)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      452..907
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(539..558)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     749..770
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    914..1035
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(925..944)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     979..998
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1158..1868)
                     /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(1436..1455)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1705..1723
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      1865..1874
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    1876..1928
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      2750..2759
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2786..2989)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2815..2835)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2990..3293
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(3468..4056)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3548..3567)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4385..4432
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4457..4476
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4664..5458)
                     /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             complement(4664..5458)
                     /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(4775..4794)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5385..5404
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(5493..5531,1..319))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(5507..5531,1..111)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 cttctggaat agctcagagg ccgaggcggc ctcggcctct gcataaataa aaaaaattag
       61 tcagccatgg ggcggagaat gggcggaact gggcggagtt aggggcggga tgggcggagt
      121 taggggcggg actatggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc
      181 tggggagcct ggggactttc cacacctggt tgctgactaa ttgagatgca tgctttgcat
      241 acttctgcct gctggggagc ctggggactt tccacaccct aactgacaca cattccacag
      301 ctggttcttt ccgcctcagg actcttcctt tttcaatatt attgaagcat ttatcagggt
      361 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
      421 ccgcgcacat ttccccgaaa agtgccacct gacgcgccct gtagcggcgc attaagcgcg
      481 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
      541 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
      601 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
      661 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
      721 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
      781 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
      841 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt
      901 acaatttacg ccttaagata cattgatgag tttggacaaa ccacaactag aatgcagtga
      961 aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc
     1021 tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag
     1081 gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtatggc tgattatgat
     1141 ctagagtcgc ggccgcttta cttgtacagc tcgtccatgc cgccggtgga gtggcggccc
     1201 tcggcgcgtt cgtactgttc cacgatggtg tagtcctcgt tgtgggaggt gatgtccaac
     1261 ttgatgttga cgttgtaggc gccgggcagc tgcacgggct tcttggcctt gtaggtggtc
     1321 ttgacctcag cgtcgtagtg gccgccgtcc ttcagcttca gcctctgctt gatctcgccc
     1381 ttcagggcgc cgtcctcggg gtacatccgc tcggaggagg cctcccagcc catggtcttc
     1441 ttctgcatta cggggccgtc ggaggggaag ttggtgccgc gcagcttcac cttgtagatg
     1501 aactcgccgt cctgcaggga ggagtcctgg gtcacggtca ccacgccgcc gtcctcgaag
     1561 ttcatcacgc gctcccactt gaagccctcg gggaaggaca gcttcaagta gtcggggatg
     1621 tcggcggggt gcttcacgta ggccttggag ccgtacatga actgagggga caggatgtcc
     1681 caggcgaagg gcagggggcc acccttggtc accttcagct tggcggtctg ggtgccctcg
     1741 taggggcggc cctcgccctc gccctcgatc tcgaactcgt ggccgttcac ggagccctcc
     1801 atgtgcacct tgaagcgcat gaactccttg atgatggcca tgttatcctc ctcgcccttg
     1861 ctcaccatgg tggcgggatc ccgggcccgc ggtaccgtcg actgcagaat tcgaagcttg
     1921 agctcgagca ggggcacaga tgtggtcgtg gtgggctcgt catagatcgg ttccatgact
     1981 ggattcacta caccggagct tccatctatc gtgtggatct gcacatgctc ctctggctca
     2041 tcaacgatct tgttataaat aaaaaatgtg acatgctcga ccccggtatc cgtggacaac
     2101 tgtgtactgt acagttgata ataatcgctt gtgaaatagc tatgcagcac cacacagtcc
     2161 ttgacaccac tctcccattt ttccgtatag cctccaatct ggtaatcatg ctcggagatt
     2221 ggtgacgttg ttccgtcacc ggaggtgatt actatggatg aggtaacgct attgtaaggt
     2281 atgcagtaat cataacaatt agtgtgccaa caaaggaagt aattggcgtc atacaagaga
     2341 ggattcttgc tccgacactt ccagcacaac cagagtctca taataattcg tacgaaattt
     2401 atgctctgga ggaagtaaac gagcgcgtac agatacagaa acggggcttc cagcccggct
     2461 gcaaccaaga gcaagtgaga atagacagtg acgaaaagaa gaagcaggtt acagacgaaa
     2521 tgaacacctt tgctgagagc caactgccat ctcttcttca gcgttatgat cttagacgct
     2581 gactggaata ccgcgagcag tgccacaccc actatgagcc agccgaatgg aagagaggct
     2641 tggatgggaa ttgtagcagt ggccctcaca aagtcactcg gggtcgcgtc cttaatttct
     2701 ccctgtttca gtgtgacagt cccgatggta aagattctca taaaaaggtc catggtggct
     2761 agcggatctg acggttcact aaaccagctc tgcttatata gacctcccac cgtacacgcc
     2821 taccgcccat ttgcgtcaat ggggcggagt tgttacgaca ttttggaaag tcccgttgat
     2881 tttggtgcca aaacaaactc ccattgacgt caatggggtg gagacttgga aatccccgtg
     2941 agtcaaaccg ctatccacgc ccattgatgt actgccaaaa ccgcatcacc atggtaatag
     3001 cgatgactaa tacgtagatg tactgccaag taggaaagtc ccataaggtc atgtactggg
     3061 cataatgcca ggcgggccat ttaccgtcat tgacgtcaat agggggcgta cttggcatat
     3121 gatacacttg atgtactgcc aagtgggcag tttaccgtaa atactccacc cattgacgtc
     3181 aatggaaagt ccctattggc gttactatgg gaacatacgt cattattgac gtcaatgggc
     3241 gggggtcgtt gggcggtcag ccaggcgggc catttaccgt aagttatgta acgcggaact
     3301 ccatatatgg gctatgaact aatgaccccg taattgatta ctattaataa ctaatgcatg
     3361 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     3421 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     3481 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     3541 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     3601 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     3661 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     3721 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     3781 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     3841 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     3901 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     3961 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     4021 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     4081 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     4141 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     4201 atatatgagt aacctgaggc tatggcaggg cctgccgccc cgacgttggc tgcgagccct
     4261 gggccttcac ccgaacttgg ggggtggggt ggggaaaagg aagaaacgcg ggcgtattgg
     4321 ccccaatggg gtctcggtgg ggtatcgaca gagtgccagc cctgggaccg aaccccgcgt
     4381 ttatgaacaa acgacccaac accgtgcgtt ttattctgtc tttttattgc cgtcatagcg
     4441 cgggttcctt ccggtattgt ctccttccgt gtttcagtta gcctccccct agggtgggcg
     4501 aagaactcca gcatgagatc cccgcgctgg aggatcatcc agccggcgtc ccggaaaacg
     4561 attccgaagc ccaacctttc atagaaggcg gcggtggaat cgaaatctcg tgatggcagg
     4621 ttgggcgtcg cttggtcggt catttcgaac cccagagtcc cgctcagaag aactcgtcaa
     4681 gaaggcgata gaaggcgatg cgctgcgaat cgggagcggc gataccgtaa agcacgagga
     4741 agcggtcagc ccattcgccg ccaagctctt cagcaatatc acgggtagcc aacgctatgt
     4801 cctgatagcg gtccgccaca cccagccggc cacagtcgat gaatccagaa aagcggccat
     4861 tttccaccat gatattcggc aagcaggcat cgccatgggt cacgacgaga tcctcgccgt
     4921 cgggcatgct cgccttgagc ctggcgaaca gttcggctgg cgcgagcccc tgatgctctt
     4981 cgtccagatc atcctgatcg acaagaccgg cttccatccg agtacgtgct cgctcgatgc
     5041 gatgtttcgc ttggtggtcg aatgggcagg tagccggatc aagcgtatgc agccgccgca
     5101 ttgcatcagc catgatggat actttctcgg caggagcaag gtgagatgac aggagatcct
     5161 gccccggcac ttcgcccaat agcagccagt cccttcccgc ttcagtgaca acgtcgagca
     5221 cagctgcgca aggaacgccc gtcgtggcca gccacgatag ccgcgctgcc tcgtcttgca
     5281 gttcattcag ggcaccggac aggtcggtct tgacaaaaag aaccgggcgc ccctgcgctg
     5341 acagccggaa cacggcggca tcagagcagc cgattgtctg ttgtgcccag tcatagccga
     5401 atagcctctc cacccaagcg gccggagaac ctgcgtgcaa tccatcttgt tcaatcatgc
     5461 gaaacgatcc tcatcctgtc tcttgatcga tctttgcaaa agcctaggcc tccaaaaaag
     5521 cctcctcact a
//
