LOCUS       sequence-325635-        6227 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 6227)
  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 6227)
  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: Nsp14D-mCherry
FEATURES             Location/Qualifiers
     source          1..6227
                     /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    1758..1810
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      1812..1821
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1818..2528
                     /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(1963..1981)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2231..2250
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2651..2772
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2688..2707)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2742..2761
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2779..3234)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2916..2937)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3128..3147
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3261..3365
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3367..3724
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3367..3558
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3390..3410)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3575..3710
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3759..4553
                     /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             3759..4553
                     /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(3813..3832)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4423..4442
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4741..4760)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4785..4832
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5161..5749
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5650..5669
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        5924..6227
                     /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 gccaccatgc
      241 aagcacctac ccacctttcc gtagatacga agttcaaaac ggaaggattg tgtgtggata
      301 taccagggat accaaaggat atgacgtacc gaaggctgat ttccatgatg ggttttaaga
      361 tgaattacca agttaatggc taccccaaca tgttcatcac cagggaggag gcaattagac
      421 acgtaagagc ctggataggc ttcgatgttg aaggttgcca tgcaacccgg gaagccgtag
      481 gcacaaacct tccgttgcag cttggctttt ccacgggcgt caacctcgtt gccgtaccga
      541 ctggctatgt tgacacgccg aacaacaccg atttctctcg cgtaagtgct aagcctcctc
      601 cgggagatca attcaagcat cttatacctc tcatgtacaa gggactgccg tggaatgtag
      661 ttcgcattaa gatcgtgcag atgctctccg atactttgaa aaatctgagc gacagggtgg
      721 tcttcgttct gtgggcacat ggcttcgaac ttacttctat gaaatacttt gtaaaaatcg
      781 ggcccgagcg gacatgttgt ttgtgcgacc gacgagctac ttgttttagc acagcatctg
      841 acacctacgc atgctggcac cacagtatag gcttcgatta cgtctacaat ccctttatga
      901 tagacgtaca acaatggggc tttacgggta acttgcagag taatcacgat ctttactgcc
      961 aagttcatgg gaacgcacac gtggcctcct gcgatgcgat aatgacgagg tgcttggccg
     1021 tgcacgagtg ctttgttaag cgggtcgatt ggactataga gtatcccata atcggtgacg
     1081 aacttaaaat taatgctgca tgcaggaaag tgcagcacat ggtagtaaaa gcagcgctcc
     1141 tcgcggataa gtttcctgtg ctccacgata tcggcaaccc taaggccatt aaatgcgtcc
     1201 cccaagctga tgtagagtgg aaattctatg acgctcagcc atgtagtgac aaagcataca
     1261 agattgaaga attgttttat tcatatgcga cacactccga taagttcact gacggagttt
     1321 gtctcttttg gaactgtaat gttgataggt atccggcgaa tagcatagtg tgtcgatttg
     1381 acacgcgcgt tctctctaat ttgaacctcc ctggctgcga cggggggtct ttgtacgtaa
     1441 acaagcatgc atttcatacc ccagcttttg acaaatccgc tttcgtaaat ctgaaacaac
     1501 ttcccttctt ctattattca gatagccctt gcgaatctca cggtaagcaa gtggttagtg
     1561 acattgacta tgtaccactt aaaagtgcaa cgtgcattac acgatgcaat ttggggggcg
     1621 cggtttgccg acaccatgcc aacgaataca gattgtacct ggatgcatat aacatgatga
     1681 taagtgcagg gtttagtctt tgggtgtata agcagttcga tacgtataat ctctggaaca
     1741 catttacccg attgcagctc gagctcaagc ttcgaattct gcagtcgacg gtaccgcggg
     1801 cccgggatcc cgccaccatg gtgagcaagg gcgaggagga taacatggcc atcatcaagg
     1861 agttcatgcg cttcaaggtg cacatggagg gctccgtgaa cggccacgag ttcgagatcg
     1921 agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg aaggtgacca
     1981 agggtggccc cctgcccttc gcctgggaca tcctgtcccc tcagttcatg tacggctcca
     2041 aggcctacgt gaagcacccc gccgacatcc ccgactactt gaagctgtcc ttccccgagg
     2101 gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc gtgacccagg
     2161 actcctccct gcaggacggc gagttcatct acaaggtgaa gctgcgcggc accaacttcc
     2221 cctccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc tccgagcgga
     2281 tgtaccccga ggacggcgcc ctgaagggcg agatcaagca gaggctgaag ctgaaggacg
     2341 gcggccacta cgacgctgag gtcaagacca cctacaaggc caagaagccc gtgcagctgc
     2401 ccggcgccta caacgtcaac atcaagttgg acatcacctc ccacaacgag gactacacca
     2461 tcgtggaaca gtacgaacgc gccgagggcc gccactccac cggcggcatg gacgagctgt
     2521 acaagtaaag cggccgcgac tctagatcat aatcagccat accacatttg tagaggtttt
     2581 acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat
     2641 tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     2701 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     2761 caatgtatct taaggcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     2821 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     2881 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     2941 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     3001 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     3061 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     3121 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     3181 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg
     3241 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     3301 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     3361 agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc
     3421 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     3481 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     3541 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     3601 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     3661 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     3721 caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     3781 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     3841 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     3901 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat
     3961 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     4021 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     4081 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     4141 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     4201 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     4261 ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc
     4321 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     4381 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     4441 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     4501 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     4561 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     4621 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     4681 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac
     4741 tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca
     4801 gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg
     4861 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt
     4921 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc
     4981 ggcaggccct gccatagcct caggttactc atatatactt tagattgatt taaaacttca
     5041 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     5101 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     5161 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     5221 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     5281 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     5341 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     5401 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     5461 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     5521 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     5581 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     5641 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     5701 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     5761 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     5821 gttatcccct gattctgtgg ataaccgtat taccgccatg cattagttat taatagtaat
     5881 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
     5941 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
     6001 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
     6061 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
     6121 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
     6181 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatg
//
