LOCUS       sequence-8865-ex        6859 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 6859)
  AUTHORS   Liou J, Kim ML, Heo WD, Jones JT, Myers JW, Ferrell JE, Meyer T
  TITLE     STIM is a Ca2+ sensor essential for Ca2+-store-depletion-triggered 
            Ca2+ influx.
  JOURNAL   Curr Biol. 2005 Jul 12. 15(13):1235-41.
  PUBMED    16005298
REFERENCE   2  (bases 1 to 6859)
  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: "Curr Biol. 
            2005 Jul 12. 15(13):1235-41."
COMMENT     Sequence Label: pEX-SP-CFP-STIM1(23-685)
FEATURES             Location/Qualifiers
     source          1..6859
                     /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 CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     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(3430..3454)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    3458..3523
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    3647..3768
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3684..3703)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3738..3757
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3775..4230)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3912..3933)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4124..4143
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4257..4361
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4363..4720
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4363..4554
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4386..4406)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4571..4706
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4755..5549
                     /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             4755..5549
                     /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(4809..4828)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5419..5438
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5737..5756)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5781..5828
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6157..6745
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6646..6665
                     /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 ccaccctgac ctggggcgtg cagtgcttca
      901 gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
      961 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
     1021 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
     1081 aggacggcaa catcctgggg cacaagctgg agtacaacta catcagccac aacgtctata
     1141 tcaccgccga caagcagaag aacggcatca aggccaactt caagatccgc cacaacatcg
     1201 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     1261 ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca
     1321 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     1381 gcatggacga gctgtacaag tccggatcaa caagtttgta caaaaaagca ggctccctca
     1441 gccatagtca cagtgagaag gcgacaggaa ccagctcggg ggccaactct gaggagtcca
     1501 ctgcagcaga gttttgccga attgacaagc ccctgtgtca cagtgaggat gagaaactca
     1561 gcttcgaggc agtccgtaac atccacaaac tgatggacga tgatgccaat ggtgatgtgg
     1621 atgtggaaga aagtgatgag ttcctgaggg aagacctcaa ttaccatgac ccaacagtga
     1681 aacacagcac cttccatggt gaggataagc tcatcagcgt ggaggacctg tggaaggcat
     1741 ggaagtcatc agaagtatac aattggaccg tggatgaggt ggtacagtgg ctgatcacat
     1801 atgtggagct gcctcagtat gaggagacct tccggaagct gcagctcagt ggccatgcca
     1861 tgccaaggct ggctgtcacc aacaccacca tgacagggac tgtgctgaag atgacagacc
     1921 ggagtcatcg gcagaagctg cagctgaagg ctctggatac agtgctcttt gggcctcctc
     1981 tcttgactcg ccataatcac ctcaaggact tcatgctggt ggtgtctatc gttattggtg
     2041 tgggcggctg ctggtttgcc tatatccaga accgttactc caaggagcac atgaagaaga
     2101 tgatgaagga cttggagggg ttacaccgag ctgagcagag tctgcatgac cttcaggaaa
     2161 ggctgcacaa ggcccaggag gagcaccgca cagtggaggt ggagaaggtc catctggaaa
     2221 agaagctgcg cgatgagatc aaccttgcta agcaggaagc ccagcggctg aaggagctgc
     2281 gggagggtac tgagaatgag cggagccgcc aaaaatatgc tgaggaggag ttggagcagg
     2341 ttcgggaggc cttgaggaaa gcagagaagg agctagaatc tcacagctca tggtatgctc
     2401 cagaggccct tcagaagtgg ctgcagctga cacatgaggt ggaggtgcaa tattacaaca
     2461 tcaagaagca aaatgctgag aagcagctgc tggtggccaa ggagggggct gagaagataa
     2521 aaaagaagag aaacacactc tttggcacct tccacgtggc ccacagctct tccctggatg
     2581 atgtagatca taaaattcta acagctaagc aagcactgag cgaggtgaca gcagcattgc
     2641 gggagcgcct gcaccgctgg caacagatcg agatcctctg tggcttccag attgtcaaca
     2701 accctggcat ccactcactg gtggctgccc tcaacataga ccccagctgg atgggcagta
     2761 cacgccccaa ccctgctcac ttcatcatga ctgacgacgt ggatgacatg gatgaggaga
     2821 ttgtgtctcc cttgtccatg cagtccccta gcctgcagag cagtgttcgg cagcgcctga
     2881 cggagccaca gcatggcctg ggatctcaga gggatttgac ccattccgat tcggagtcct
     2941 ccctccacat gagtgaccgc cagcgtgtgg cccccaaacc tcctcagatg agccgtgctg
     3001 cagacgaggc tctcaatgcc atgacttcca atggcagcca ccggctgatc gagggggtcc
     3061 acccagggtc tctggtggag aaactgcctg acagccctgc cctggccaag aaggcattac
     3121 tggcgctgaa ccatgggctg gacaaggccc acagcctgat ggagctgagc ccctcagccc
     3181 cacctggtgg ctctccacat ttggattctt cccgttctca cagccccagc tccccagacc
     3241 cagacacacc atctccagtt ggggacagcc gagccctgca agccagccga aacacacgca
     3301 ttccccacct ggctggcaag aaggctgtgg ctgaggagga taatggctct attggcgagg
     3361 aaacagactc cagcccaggc cggaagaagt ttcctctcaa aatctttaag aagcctctta
     3421 agaagtagaa cccagctttc ttgtacaaag tggttgatcc ggactcagat ctcgagctca
     3481 agcttcgaat tctgcagtcg acggtaccgc gggcccggga tccaccggat ctagataact
     3541 gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca
     3601 cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc
     3661 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     3721 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaac gcgtaaattg
     3781 taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta
     3841 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt
     3901 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
     3961 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa
     4021 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
     4081 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
     4141 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
     4201 ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg
     4261 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     4321 aaccctgata aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca
     4381 gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     4441 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     4501 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     4561 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     4621 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     4681 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat
     4741 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     4801 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     4861 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     4921 ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     4981 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     5041 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     5101 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     5161 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     5221 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     5281 cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     5341 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     5401 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     5461 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     5521 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca
     5581 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     5641 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     5701 gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc
     5761 ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc
     5821 gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag
     5881 accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc
     5941 gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg
     6001 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     6061 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     6121 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     6181 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     6241 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     6301 tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     6361 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     6421 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     6481 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     6541 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     6601 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     6661 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     6721 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     6781 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     6841 ccgtattacc gccatgcat
//
