LOCUS       sequence-417516-        7607 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses STIM1 fused with 2Strep in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7607)
  AUTHORS   Li J, Gui Q, Liang FX, Sall J, Zhang Q, Duan Y, Dhabaria A, Askenazi
            M, Ueberheide B, Stapleford KA, Pagano M
  TITLE     The REEP5/TRAM1 complex binds SARS-CoV-2 NSP3 and promotes virus 
            replication.
  JOURNAL   J Virol. 2023 Sep 28:e0050723. doi: 10.1128/jvi.00507-23.
  PUBMED    37768083
REFERENCE   2  (bases 1 to 7607)
  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"; journalName: "J Virol. 
            2023 Sep 28:e0050723. doi: 10.1128/jvi.00507-23."
COMMENT     Sequence Label: STIM1-2Strep
FEATURES             Location/Qualifiers
     source          1..7607
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(38..57)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        229..608
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        609..812
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     763..783
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     857..876
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        857..875
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2971..2980
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      2975..2984
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2992..3015
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             3052..3075
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             3100..3123
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     promoter        complement(3154..3172)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3155..3172)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3192..3209)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3198..3422
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3468..3896
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3555..3574)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3765..3786
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3905..3925)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3910..4239
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4090..4225
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4152..4171
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4306..5100
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4360..4379)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4970..4989
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5274..5395
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5311..5330)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5365..5384
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5444..5460)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5444..5460)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5457..5479)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5468..5484
                     /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(5492..5498,5499..5516,5517..5522))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5537..5558
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5675..5692)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5846..6434)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5926..5945)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6605..7396,7397..7465))
                     /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     7228..7247
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7466..7570)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcgggagatc tcccgatccc ctatggtgca ctctcagtac aatctgctct gatgccgcat
       61 agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca
      121 aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttaggg
      181 ttaggcgttt tgcgctgctt cgcgatgtac gggccagata tacgcgttga cattgattat
      241 tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
      301 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
      361 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
      421 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
      481 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
      541 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
      601 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
      661 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
      721 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
      781 gtgtacggtg ggaggtctat ataagcagag ctctctggct aactagagaa cccactgctt
      841 actggcttat cgaaattaat acgactcact atagggagac ccaagcttgg taccgagctc
      901 ggatccatgg atgtatgcgt ccgtcttgcc ctgtggctcc tctggggact cctcctgcac
      961 cagggccaga gcctcagcca tagtcacagt gagaaggcga caggaaccag ctcgggggcc
     1021 aactctgagg agtccactgc agcagagttt tgccgaattg acaagcccct gtgtcacagt
     1081 gaggatgaga aactcagctt cgaggcagtc cgtaacatcc acaaactgat ggacgatgat
     1141 gccaatggtg atgtggatgt ggaagaaagt gatgagttcc tgagggaaga cctcaattac
     1201 catgacccaa cagtgaaaca cagcaccttc catggtgagg ataagctcat cagcgtggag
     1261 gacctgtgga aggcatggaa gtcatcagaa gtatacaatt ggaccgtgga tgaggtggta
     1321 cagtggctga tcacatatgt ggagctgcct cagtatgagg agaccttccg gaagctgcag
     1381 ctcagtggcc atgccatgcc aaggctggct gtcaccaaca ccaccatgac agggactgtg
     1441 ctgaagatga cagaccggag tcatcggcag aagctgcagc tgaaggctct ggatacagtg
     1501 ctctttgggc ctcctctctt gactcgccat aatcacctca aggacttcat gctggtggtg
     1561 tctatcgtta ttggtgtggg cggctgctgg tttgcctata tccagaaccg ttactccaag
     1621 gagcacatga agaagatgat gaaggacttg gaggggttac accgagctga gcagagtctg
     1681 catgaccttc aggaaaggct gcacaaggcc caggaggagc accgcacagt ggaggtggag
     1741 aaggtccatc tggaaaagaa gctgcgcgat gagatcaacc ttgctaagca ggaagcccag
     1801 cggctgaagg agctgcggga gggtactgag aatgagcgga gccgccaaaa atatgctgag
     1861 gaggagttgg agcaggttcg ggaggccttg aggaaagcag agaaggagct agaatctcac
     1921 agctcatggt atgctccaga ggcccttcag aagtggctgc agctgacaca tgaggtggag
     1981 gtgcaatatt acaacatcaa gaagcaaaat gctgagaagc agctgctggt ggccaaggag
     2041 ggggctgaga agataaaaaa gaagagaaac acactctttg gcaccttcca cgtggcccac
     2101 agctcttccc tggatgatgt agatcataaa attctaacag ctaagcaagc actgagcgag
     2161 gtgacagcag cattgcggga gcgcctgcac cgctggcaac agatcgagat cctctgtggc
     2221 ttccagattg tcaacaaccc tggcatccac tcactggtgg ctgccctcaa catagacccc
     2281 agctggatgg gcagtacacg ccccaaccct gctcacttca tcatgactga cgacgtggat
     2341 gacatggatg aggagattgt gtctcccttg tccatgcagt cccctagcct gcagagcagt
     2401 gttcggcagc gcctgacgga gccacagcat ggcctgggat ctcagaggga tttgacccat
     2461 tccgattcgg agtcctccct ccacatgagt gaccgccagc gtgtggcccc caaacctcct
     2521 cagatgagcc gtgctgcaga cgaggctctc aatgccatga cttccaatgg cagccaccgg
     2581 ctgatcgagg gggtccaccc agggtctctg gtggagaaac tgcctgacag ccctgccctg
     2641 gccaagaagg cattactggc gctgaaccat gggctggaca aggcccacag cctgatggag
     2701 ctgagcccct cagccccacc tggtggctct ccacatttgg attcttcccg ttctcacagc
     2761 cccagctccc cagacccaga cacaccatct ccagttgggg acagccgagc cctgcaagcc
     2821 agccgaaaca cacgcattcc ccacctggct ggcaagaagg ctgtggctga ggaggataat
     2881 ggctctattg gcgaggaaac agactccagc ccaggccgga agaagtttcc cctcaaaatc
     2941 tttaagaagc ctcttaagaa ggaattcgcc gccaccatgg cggccgccag ctggagccac
     3001 cctcagttcg agaagggagg aggaagcggc ggaggcagcg gaggaggaag ctggagccac
     3061 ccgcagttcg agaaatgact cgagcatgca tctagaggct ggagccaccc gcagttcgag
     3121 aaatgactcg agcatgcatc tagagggccc tattctatag tgtcacctaa atgctagagc
     3181 tcgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     3241 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     3301 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     3361 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     3421 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
     3481 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     3541 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     3601 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     3661 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     3721 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     3781 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     3841 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     3901 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     3961 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     4021 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     4081 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     4141 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     4201 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     4261 tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     4321 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     4381 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     4441 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     4501 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     4561 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     4621 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     4681 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     4741 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     4801 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     4861 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     4921 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     4981 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     5041 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     5101 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     5161 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     5221 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
     5281 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     5341 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     5401 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     5461 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     5521 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     5581 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     5641 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     5701 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     5761 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     5821 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     5881 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     5941 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     6001 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     6061 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     6121 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     6181 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     6241 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     6301 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     6361 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     6421 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     6481 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     6541 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     6601 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     6661 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     6721 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     6781 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     6841 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     6901 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     6961 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     7021 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     7081 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     7141 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     7201 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     7261 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     7321 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     7381 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     7441 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     7501 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     7561 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacgga
//
