LOCUS       sequence-325887-        4958 bp    DNA     circular SYN 14-APR-2025
DEFINITION  mammalian expression and localization.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4958)
  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 4958)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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: Nsp7-mCherry
FEATURES             Location/Qualifiers
     source          1..4958
                     /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    489..541
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      543..552
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             549..1259
                     /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(694..712)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     962..981
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    1382..1503
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1419..1438)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1473..1492
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1510..1965)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1647..1668)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1859..1878
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1992..2096
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2098..2455
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2098..2289
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2121..2141)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2306..2441
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2490..3284
                     /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             2490..3284
                     /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(2544..2563)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3154..3173
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3472..3491)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3516..3563
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3892..4480
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4381..4400
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        4655..4958
                     /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 gcaagatgag cgacgtaaaa tgcacgtcag ttgttctctt gagcgtcctg caacaactca
      301 gagtggagtc tagttccaag ctctgggctc aatgcgttca gcttcacaat gacattctgt
      361 tggcgaaaga taccacggag gcattcgaga agatggtctc acttctgagc gttctcctgt
      421 caatgcaagg ggcggtcgat atcaataaat tgtgtgagga aatgctcgac aaccgggcga
      481 cattgcagct cgagctcaag cttcgaattc tgcagtcgac ggtaccgcgg gcccgggatc
      541 ccgccaccat ggtgagcaag ggcgaggagg ataacatggc catcatcaag gagttcatgc
      601 gcttcaaggt gcacatggag ggctccgtga acggccacga gttcgagatc gagggcgagg
      661 gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggtggcc
      721 ccctgccctt cgcctgggac atcctgtccc ctcagttcat gtacggctcc aaggcctacg
      781 tgaagcaccc cgccgacatc cccgactact tgaagctgtc cttccccgag ggcttcaagt
      841 gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac cgtgacccag gactcctccc
      901 tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg caccaacttc ccctccgacg
      961 gccccgtaat gcagaagaag accatgggct gggaggcctc ctccgagcgg atgtaccccg
     1021 aggacggcgc cctgaagggc gagatcaagc agaggctgaa gctgaaggac ggcggccact
     1081 acgacgctga ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg cccggcgcct
     1141 acaacgtcaa catcaagttg gacatcacct cccacaacga ggactacacc atcgtggaac
     1201 agtacgaacg cgccgagggc cgccactcca ccggcggcat ggacgagctg tacaagtaaa
     1261 gcggccgcga ctctagatca taatcagcca taccacattt gtagaggttt tacttgcttt
     1321 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
     1381 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     1441 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     1501 ttaaggcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     1561 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
     1621 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     1681 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     1741 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     1801 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     1861 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     1921 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc
     1981 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     2041 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg
     2101 aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     2161 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     2221 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     2281 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     2341 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     2401 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg
     2461 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     2521 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     2581 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     2641 ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg
     2701 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     2761 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     2821 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     2881 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     2941 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     3001 tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     3061 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     3121 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     3181 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     3241 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     3301 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     3361 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     3421 gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg
     3481 gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac
     3541 gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt
     3601 cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc
     3661 ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc
     3721 tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt
     3781 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     3841 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     3901 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     3961 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     4021 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     4081 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     4141 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     4201 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     4261 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     4321 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     4381 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     4441 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     4501 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     4561 tgattctgtg gataaccgta ttaccgccat gcattagtta ttaatagtaa tcaattacgg
     4621 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
     4681 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
     4741 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
     4801 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
     4861 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
     4921 ggcagtacat ctacgtatta gtcatcgcta ttaccatg
//
