LOCUS       sequence-417517-        6749 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses STIM1 fused with EGFP in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6749)
  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 6749)
  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-EGFP
FEATURES             Location/Qualifiers
     source          1..6749
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        62..365
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        366..569
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     520..540
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2690..2699
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2696..2698,2699..2701,2702..3415)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2741..2762)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3002..3021)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3349..3370
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3538..3659
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3575..3594)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3629..3648
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3666..4121)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3803..3824)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4015..4034
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4148..4252
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4254..4611
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4254..4445
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4277..4297)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4462..4597
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4646..5440
                     /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             4646..5440
                     /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(4700..4719)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5310..5329
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5628..5647)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5672..5719
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6048..6636
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6537..6556
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc
       61 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
      121 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
      181 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
      241 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
      301 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
      361 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
      421 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
      481 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
      541 tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgctagcgct
      601 accggactca gatctcgaga tggatgtatg cgtccgtctt gccctgtggc tcctctgggg
      661 actcctcctg caccagggcc agagcctcag ccatagtcac agtgagaagg cgacaggaac
      721 cagctcgggg gccaactctg aggagtccac tgcagcagag ttttgccgaa ttgacaagcc
      781 cctgtgtcac agtgaggatg agaaactcag cttcgaggca gtccgtaaca tccacaaact
      841 gatggacgat gatgccaatg gtgatgtgga tgtggaagaa agtgatgagt tcctgaggga
      901 agacctcaat taccatgacc caacagtgaa acacagcacc ttccatggtg aggataagct
      961 catcagcgtg gaggacctgt ggaaggcatg gaagtcatca gaagtataca attggaccgt
     1021 ggatgaggtg gtacagtggc tgatcacata tgtggagctg cctcagtatg aggagacctt
     1081 ccggaagctg cagctcagtg gccatgccat gccaaggctg gctgtcacca acaccaccat
     1141 gacagggact gtgctgaaga tgacagaccg gagtcatcgg cagaagctgc agctgaaggc
     1201 tctggataca gtgctctttg ggcctcctct cttgactcgc cataatcacc tcaaggactt
     1261 catgctggtg gtgtctatcg ttattggtgt gggcggctgc tggtttgcct atatccagaa
     1321 ccgttactcc aaggagcaca tgaagaagat gatgaaggac ttggaggggt tacaccgagc
     1381 tgagcagagt ctgcatgacc ttcaggaaag gctgcacaag gcccaggagg agcaccgcac
     1441 agtggaggtg gagaaggtcc atctggaaaa gaagctgcgc gatgagatca accttgctaa
     1501 gcaggaagcc cagcggctga aggagctgcg ggagggtact gagaatgagc ggagccgcca
     1561 aaaatatgct gaggaggagt tggagcaggt tcgggaggcc ttgaggaaag cagagaagga
     1621 gctagaatct cacagctcat ggtatgctcc agaggccctt cagaagtggc tgcagctgac
     1681 acatgaggtg gaggtgcaat attacaacat caagaagcaa aatgctgaga agcagctgct
     1741 ggtggccaag gagggggctg agaagataaa aaagaagaga aacacactct ttggcacctt
     1801 ccacgtggcc cacagctctt ccctggatga tgtagatcat aaaattctaa cagctaagca
     1861 agcactgagc gaggtgacag cagcattgcg ggagcgcctg caccgctggc aacagatcga
     1921 gatcctctgt ggcttccaga ttgtcaacaa ccctggcatc cactcactgg tggctgccct
     1981 caacatagac cccagctgga tgggcagtac acgccccaac cctgctcact tcatcatgac
     2041 tgacgacgtg gatgacatgg atgaggagat tgtgtctccc ttgtccatgc agtcccctag
     2101 cctgcagagc agtgttcggc agcgcctgac ggagccacag catggcctgg gatctcagag
     2161 ggatttgacc cattccgatt cggagtcctc cctccacatg agtgaccgcc agcgtgtggc
     2221 ccccaaacct cctcagatga gccgtgctgc agacgaggct ctcaatgcca tgacttccaa
     2281 tggcagccac cggctgatcg agggggtcca cccagggtct ctggtggaga aactgcctga
     2341 cagccctgcc ctggccaaga aggcattact ggcgctgaac catgggctgg acaaggccca
     2401 cagcctgatg gagctgagcc cctcagcccc acctggtggc tctccacatt tggattcttc
     2461 ccgttctcac agccccagct ccccagaccc agacacacca tctccagttg gggacagccg
     2521 agccctgcaa gccagccgaa acacacgcat tccccacctg gctggcaaga aggctgtggc
     2581 tgaggaggat aatggctcta ttggcgagga aacagactcc agcccaggcc ggaagaagtt
     2641 tcccctcaaa atctttaaga agcctcttaa gaaggcggat ccaccggtcg ccaccatggt
     2701 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
     2761 cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa
     2821 gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
     2881 gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca
     2941 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa
     3001 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
     3061 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
     3121 ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat
     3181 caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca
     3241 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct
     3301 gagcacccag tccaagctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct
     3361 ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca agtaaagcgg
     3421 ccgcgactct agatcataat cagccatacc acatttgtag aggttttact tgctttaaaa
     3481 aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac
     3541 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     3601 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa
     3661 ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     3721 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     3781 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     3841 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     3901 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     3961 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     4021 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     4081 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg
     4141 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     4201 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc
     4261 ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     4321 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     4381 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     4441 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     4501 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     4561 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca
     4621 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
     4681 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     4741 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     4801 cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac
     4861 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     4921 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     4981 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     5041 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     5101 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     5161 caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     5221 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     5281 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     5341 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     5401 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
     5461 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
     5521 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     5581 ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag
     5641 gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac
     5701 ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat
     5761 accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac
     5821 cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc
     5881 atagcctcag gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     5941 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     6001 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     6061 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     6121 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     6181 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     6241 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     6301 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     6361 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     6421 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     6481 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     6541 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     6601 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     6661 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     6721 tctgtggata accgtattac cgccatgca
//
