LOCUS       sequence-8871-ex        6928 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6928)
  AUTHORS   Brandman O, Liou J, Park WS, Meyer T
  TITLE     STIM2 is a feedback regulator that stabilizes basal cytosolic and 
            endoplasmic reticulum Ca2+ levels.
  JOURNAL   Cell. 2007 Dec 28. 131(7):1327-39.
  PUBMED    18160041
REFERENCE   2  (bases 1 to 6928)
  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: "Cell. 2007 
            Dec 28. 131(7):1327-39."
COMMENT     Sequence Label: pEX-CMV-SP-YFP-STM1(24-685) EF hand 3 mut (A79...
FEATURES             Location/Qualifiers
     source          1..6928
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      678..687
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(684..686,687..689,690..1400)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(729..750)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(990..1009)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1337..1358
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    1410..1434
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3499..3523)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    3527..3592
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    3716..3837
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3753..3772)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3807..3826
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3844..4299)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3981..4002)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4193..4212
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4326..4430
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4432..4789
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4432..4623
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4455..4475)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4640..4775
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4824..5618
                     /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             4824..5618
                     /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(4878..4897)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5488..5507
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5806..5825)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5850..5897
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6226..6814
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6715..6734
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcacca
      601 tggatgtatg cgtccgtctt gccctgtggc tcctctgggg actcctcctg caccagggcc
      661 agagcctcgc accggtcgcc accatggtga gcaagggcga ggagctgttc accggggtgg
      721 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
      781 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca
      841 agctgcccgt gccctggccc accctcgtga ccaccttcgg ctacggcctg cagtgcttcg
      901 cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
      961 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
     1021 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
     1081 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata
     1141 tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg
     1201 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     1261 ccgtgctgct gcccgacaac cactacctga gctaccagtc cgccctgagc aaagacccca
     1321 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     1381 gcatggacga gctgtacaag tccggatcaa caagtttgta caaaaaagca ggctccctca
     1441 tggatgtatg cgtccgtctt gccctgtggc tcctctgggg gctcctcctg caccagggcc
     1501 agagcctcag ccatagtcac agtgagaagg cgacaggaac cagctcgggg gccaactctg
     1561 aggagtccac tgcagcagag ttttgccgaa ttgacaagcc cctgtgtcac agtgaggatg
     1621 agaaactcag cttcgaggca gtccgtaaca tccacaaact gatggacgat gataaagatg
     1681 gtggtgtgga tgtggaagaa agtgatgagt tcctgaggga agacctcaat taccatgacc
     1741 caacagtgaa acacagcacc ttccatggtg aggataagct catcagcgtg gaggacctgt
     1801 ggaaggcatg gaagtcatca gaagtataca attggaccgt ggatgaggtg gtacagtggc
     1861 tgatcacata tgtggagctg cctcagtatg aggagacctt ccggaagctg cagctcagtg
     1921 gccatgccat gccaaggctg gctgtcacca acaccaccat gacagggact gtgctgaaga
     1981 tgacagaccg gagtcatcgg cagaagctgc agctgaaggc tctggataca gtgctctttg
     2041 ggcctcctct cttgactcgc cataatcacc tcaaggactt catgctggtg gtgtctatcg
     2101 ttattggtgt gggcggctgc tggtttgcct atatccagaa ccgttactcc aaggagcaca
     2161 tgaagaagat gatgaaggac ttggaggggt tacaccgagc tgagcagagt ctgcatgacc
     2221 ttcaggaaag gctgcacaag gcccaggagg agcaccgcac agtggaggtg gagaaggtcc
     2281 atctggaaaa gaagctgcgc gatgagatca accttgctaa gcaggaagcc cagcggctga
     2341 aggagctgcg ggagggtact gagaatgagc ggagccgcca aaaatatgct gaggaggagt
     2401 tggagcaggt tcgggaggcc ttgaggaaag cagagaagga gctagaatct cacagctcat
     2461 ggtatgctcc agaggccctt cagaagtggc tgcagctgac acatgaggtg gaggtgcaat
     2521 attacaacat caagaagcaa aatgctgaga agcagctgct ggtggccaag gagggggctg
     2581 agaagataaa aaagaagaga aacacactct ttggcacctt ccacgtggcc cacagctctt
     2641 ccctggatga tgtagatcat aaaattctaa cagctaagca agcactgagc gaggtgacag
     2701 cagcattgcg ggagcgcctg caccgctggc aacagatcga gatcctctgt ggcttccaga
     2761 ttgtcaacaa ccctggcatc cactcactgg tggctgccct caacatagac cccagctgga
     2821 tgggcagtac acgccccaac cctgctcact tcatcatgac tgacgacgtg gatgacatgg
     2881 atgaggagat tgtgtctccc ttgtccatgc agtcccctag cctgcagagc agtgttcggc
     2941 agcgcctgac ggagccacag catggcctgg gatctcagag ggatttgacc cattccgatt
     3001 cggagtcctc cctccacatg agtgaccgcc agcgtgtggc ccccaaacct cctcagatga
     3061 gccgtgctgc agacgaggct ctcaatgcca tgacttccaa tggacgccac cggctgatcg
     3121 agggggtcca cccagggtct ctggtggaga aactgcctga cagccctgcc ctggccaaga
     3181 aggcattact ggcgctgaac catgggctgg acaaggccca cagcctgatg gagctgagcc
     3241 cctcagcccc acctggtggc tctccacatt tggattcttc ccgttctcac agccccagct
     3301 ccccagaccc agacacacca tctccagttg gggacagccg agccctgcaa gccagccgaa
     3361 acacacgcat tccccacctg gctggcaaga aggctgtggc tgaggaggat aatggctcta
     3421 ttggcgagga aacagactcc agcccaggcc ggaagaagtt tcctctcaaa atctttaaga
     3481 agcctcttaa gaagtagaac ccagctttct tgtacaaagt ggttgatccg gactcagatc
     3541 tcgagctcaa gcttcgaatt ctgcagtcga cggtaccgcg ggcccgggat ccaccggatc
     3601 tagataactg atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa
     3661 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
     3721 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     3781 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaacg
     3841 cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     3901 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     3961 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     4021 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     4081 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     4141 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     4201 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     4261 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa
     4321 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     4381 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg
     4441 aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     4501 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     4561 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     4621 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     4681 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     4741 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag
     4801 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     4861 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     4921 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     4981 tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga
     5041 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     5101 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     5161 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     5221 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     5281 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     5341 ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     5401 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     5461 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     5521 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     5581 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     5641 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     5701 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     5761 ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga
     5821 gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg
     5881 tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac
     5941 cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc
     6001 cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat
     6061 agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     6121 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     6181 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     6241 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     6301 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     6361 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     6421 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     6481 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     6541 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     6601 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     6661 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     6721 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     6781 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     6841 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     6901 tgtggataac cgtattaccg ccatgcat
//
