LOCUS       sequence-454347-        8270 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral expression of mCherry-STIM1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8270)
  AUTHORS   Zhao G, Liu S, Arun S, Renda F, Khodjakov A, Pellman D
  TITLE     A tubule-sheet continuum model for the mechanism of nuclear envelope
            assembly.
  JOURNAL   Dev Cell. 2023 May 22;58(10):847-865.e10. doi: 
            10.1016/j.devcel.2023.04.003. Epub 2023 Apr 24.
  PUBMED    37098350
REFERENCE   2  (bases 1 to 8270)
  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: 
            "10.1016/j.devcel.2023.04.003"; journalName: "Dev Cell"; date: 
            "2023-05-22- 22"; volume: "58"; issue: "10"; pages: "847-865.e10"
COMMENT     Sequence Label: mCherry-STIM1
FEATURES             Location/Qualifiers
     source          1..8270
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    40..56
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(64..70,71..88,89..94))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    109..130
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(247..264)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(418..1006)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(498..517)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1177..1968,1969..2037))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1800..1819
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2038..2142)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2210..2228
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2266..2288)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2388..2407
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2552..2931
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2932..3135
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3086..3106
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    3375..3716
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             3783..4199
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     4129..4151
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     4167..4183
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      6318..6327
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6324..7034
                     /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(6469..6487)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     6737..6756
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             7045..7068
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             7092..7118
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             7119..7145
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(7278..7301)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     LTR             7634..8148
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
ORIGIN
        1 ttatggacag cgcagaaaga gctggggaga attgtgaaat tgttatccgc tcacaattcc
       61 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
      121 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
      181 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
      241 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
      301 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
      361 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
      421 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
      481 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
      541 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
      601 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
      661 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
      721 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
      781 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
      841 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
      901 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
      961 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     1021 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     1081 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     1141 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     1201 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     1261 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     1321 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     1381 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     1441 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     1501 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     1561 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     1621 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     1681 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     1741 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     1801 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     1861 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     1921 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     1981 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     2041 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     2101 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     2161 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     2221 cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     2281 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     2341 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
     2401 gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa
     2461 ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
     2521 gcgggcctct tcgctattac gccagcgcgt tgacattgat tattgactag ttattaatag
     2581 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
     2641 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
     2701 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
     2761 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct
     2821 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg
     2881 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg
     2941 ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc
     3001 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa
     3061 tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc
     3121 tatataagca gagctcaata aaagagccca caacccctca ctcggcgcgc cagtcctccg
     3181 atagactgcg tcgcccgggt acccgtattc ccaataaagc ctcttgctgt ttgcatccga
     3241 atcgtggact cgctgatcct tgggagggtc tcctcagatt gattgactgc ccacctcggg
     3301 ggtctttcat ttggaggttc caccgagatt tggagacccc tgcccaggga ccaccgaccc
     3361 ccccgccggg aggtaagctg gccagcggtc gtttcgtgtc tgtctctgtc tttgtgcgtg
     3421 tttgtgccgg catctaatgt ttgcgcctgc gtctgtacta gttagctaac tagctctgta
     3481 tctggcggac ccgtggtgga actgacgagt tctgaacacc cggccgcaac cctgggagac
     3541 gtcccaggga ctttgggggc cgtttttgtg gcccgacctg aggaagggag tcgatgtgga
     3601 atccgacccc gtcaggatat gtggttctgg taggagacga gaacctaaaa cagttcccgc
     3661 ctccgtctga atttttgctt tcggtttgga accgaagccg cgcgtcttgt ctgctgcagc
     3721 gctgcagcat cgttctgtgt tgtctctgtc tgactgtgtt tctgtatttg tctgaaaatt
     3781 agggccagac tgttaccact cccttaagtt tgaccttagg tcactggaaa gatgtcgagc
     3841 ggatcgctca caaccagtcg gtagatgtca agaagagacg ttgggttacc ttctgctctg
     3901 cagaatggcc aacctttaac gtcggatggc cgcgagacgg cacctttaac cgagacctca
     3961 tcacccaggt taagatcaag gtcttttcac ctggcccgca tggacaccca gaccaggtcc
     4021 cctacatcgt gacctgggaa gccttggctt ttgacccccc tccctgggtc aagccctttg
     4081 tacaccctaa gcctccgcct cctcttcctc catccgcccc gtctctcccc cttgaacctc
     4141 ctcgttcgac cccgcctcga tcctcccttt atccagccct cactccttct ctaggcgccg
     4201 gaattagatc tctcgagatg gatgtatgcg tccgtcttgc cctgtggctc ctctggggac
     4261 tcctcctgca ccagggccag agcctcagcc atagtcacag tgagaaggcg acaggaacca
     4321 gctcgggggc caactctgag gagtccactg cagcagagtt ttgccgaatt gacaagcccc
     4381 tgtgtcacag tgaggatgag aaactcagct tcgaggcagt ccgtaacatc cacaaactga
     4441 tggacgatga tgccaatggt gatgtggatg tggaagaaag tgatgagttc ctgagggaag
     4501 acctcaatta ccatgaccca acagtgaaac acagcacctt ccatggtgag gataagctca
     4561 tcagcgtgga ggacctgtgg aaggcatgga agtcatcaga agtatacaat tggaccgtgg
     4621 atgaggtggt acagtggctg atcacatatg tggagctgcc tcagtatgag gagaccttcc
     4681 ggaagctgca gctcagtggc catgccatgc caaggctggc tgtcaccaac accaccatga
     4741 cagggactgt gctgaagatg acagaccgga gtcatcggca gaagctgcag ctgaaggctc
     4801 tggatacagt gctctttggg cctcctctct tgactcgcca taatcacctc aaggacttca
     4861 tgctggtggt gtctatcgtt attggtgtgg gcggctgctg gtttgcctat atccagaacc
     4921 gttactccaa ggagcacatg aagaagatga tgaaggactt ggaggggtta caccgagctg
     4981 agcagagtct gcatgacctt caggaaaggc tgcacaaggc ccaggaggag caccgcacag
     5041 tggaggtgga gaaggtccat ctggaaaaga agctgcgcga tgagatcaac cttgctaagc
     5101 aggaagccca gcggctgaag gagctgcggg agggtactga gaatgagcgg agccgccaaa
     5161 aatatgctga ggaggagttg gagcaggttc gggaggcctt gaggaaagca gagaaggagc
     5221 tagaatctca cagctcatgg tatgctccag aggcccttca gaagtggctg cagctgacac
     5281 atgaggtgga ggtgcaatat tacaacatca agaagcaaaa tgctgagaag cagctgctgg
     5341 tggccaagga gggggctgag aagataaaaa agaagagaaa cacactcttt ggcaccttcc
     5401 acgtggccca cagctcttcc ctggatgatg tagatcataa aattctaaca gctaagcaag
     5461 cactgagcga ggtgacagca gcattgcggg agcgcctgca ccgctggcaa cagatcgaga
     5521 tcctctgtgg cttccagatt gtcaacaacc ctggcatcca ctcactggtg gctgccctca
     5581 acatagaccc cagctggatg ggcagtacac gccccaaccc tgctcacttc atcatgactg
     5641 acgacgtgga tgacatggat gaggagattg tgtctccctt gtccatgcag tcccctagcc
     5701 tgcagagcag tgttcggcag cgcctgacgg agccacagca tggcctggga tctcagaggg
     5761 atttgaccca ttccgattcg gagtcctccc tccacatgag tgaccgccag cgtgtggccc
     5821 ccaaacctcc tcagatgagc cgtgctgcag acgaggctct caatgccatg acttccaatg
     5881 gcagccaccg gctgatcgag ggggtccacc cagggtctct ggtggagaaa ctgcctgaca
     5941 gccctgccct ggccaagaag gcattactgg cgctgaacca tgggctggac aaggcccaca
     6001 gcctgatgga gctgagcccc tcagccccac ctggtggctc tccacatttg gattcttccc
     6061 gttctcacag ccccagctcc ccagacccag acacaccatc tccagttggg gacagccgag
     6121 ccctgcaagc cagccgaaac acacgcattc cccacctggc tggcaagaag gctgtggctg
     6181 aggaggataa tggctctatt ggcgaggaaa cagactccag cccaggccgg aagaagtttc
     6241 ccctcaaaat ctttaagaag cctcttaaga agcgaattct gcagtcgacg gtaccgcggg
     6301 cccgggatcc accggtcgcc accatggtga gcaagggcga ggaggataac atggccatca
     6361 tcaaggagtt catgcgcttc aaggtgcaca tggagggctc cgtgaacggc cacgagttcg
     6421 agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg
     6481 tgaccaaggg tggccccctg cccttcgcct gggacatcct gtcccctcag ttcatgtacg
     6541 gctccaaggc ctacgtgaag caccccgccg acatccccga ctacttgaag ctgtccttcc
     6601 ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga
     6661 cccaggactc ctccctgcag gacggcgagt tcatctacaa ggtgaagctg cgcggcacca
     6721 acttcccctc cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctcctccg
     6781 agcggatgta ccccgaggac ggcgccctga agggcgagat caagcagagg ctgaagctga
     6841 aggacggcgg ccactacgac gctgaggtca agaccaccta caaggccaag aagcccgtgc
     6901 agctgcccgg cgcctacaac gtcaacatca agttggacat cacctcccac aacgaggact
     6961 acaccatcgt ggaacagtac gaacgcgccg agggccgcca ctccaccggc ggcatggacg
     7021 agctgtacaa gtaaagcggc cgctgactac aaggacgacg atgacaagtg agaattgtcg
     7081 aacgggccgc ttacccatac gatgttccgg attacgctta cccatatgat gttccggatt
     7141 acgcttaact agagtgaggg tccccacctg ggacccttga gagtatcagg tctcccacgt
     7201 gggagacaag aaatccctgt ttaatattta acgaattcta ccgggtaggg gaggcgcttt
     7261 tcccaaggca gtctggagca tgcgctttag cagccccgct gggcacttgg cgctacacaa
     7321 gtggcctctg gcctcgcaca cattccacat ccaccggtag gcgccaaccg gctccgttct
     7381 ttggtggccc cttcgcgcca ccttctactc ctcccctagt caggaagttc ccccccgccc
     7441 cgcagctcgc gtcgtgcagg acgtgacaaa tggaagtagc acgtctcact agtctcgtgc
     7501 agatggacag caccgctgag caatggaagc gggtaggcct ttggggcagc ggccaagccc
     7561 gaccgaaagg agcgcacgac cccatgcatc gataaaataa aagattttat ttagtctcca
     7621 gaaaaagggg ggaatgaaag accccacctg taggtttggc aagctagctt aagtaacgcc
     7681 attttgcaag gcatggaaaa tacataactg agaatagaga agttcagatc aaggttagga
     7741 acagagagac agcagaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg
     7801 gctcagggcc aagaacagat ggtccccaga tgcggtcccg ccctcagcag tttctagaga
     7861 accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc ttatttgaac
     7921 taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa
     7981 agagcccaca acccctcact cggcgcgcca gtcctccgat agactgcgtc gcccgggtac
     8041 ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc tgttccttgg
     8101 gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcatgg gtaacagttt
     8161 cttgaagttg gagaacaaca ttctgagggt aggagtcgaa tattaagtaa tcctgactca
     8221 attagccact gttttgaatc cacatactcc aatactcctg aaatagttca
//
