LOCUS       sequence-8869-ex        7069 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 7069)
  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 7069)
  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-CFP-STM2(15-746)
FEATURES             Location/Qualifiers
     source          1..7069
                     /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(3640..3664)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    3668..3733
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    3857..3978
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3894..3913)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3948..3967
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3985..4440)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4122..4143)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4334..4353
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4467..4571
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4573..4930
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4573..4764
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4596..4616)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4781..4916
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4965..5759
                     /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             4965..5759
                     /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(5019..5038)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5629..5648
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5947..5966)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5991..6038
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6367..6955
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6856..6875
                     /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 ggctccctct
     1441 gcgagcttgt gccccggcac ctccgcgggc ggcgggcgac tggctctgcc gcaactgccg
     1501 cctcctctct cgccgcggcg gccggcgata gcccggcgct catgacagat ccctgcatgt
     1561 cactgagtcc accatgcttt acagaagaag acagatttag tctggaagct cttcaaacaa
     1621 tacataaaca aatggatgat gacaaagatg gtggaattga agtagaggaa agtgatgaat
     1681 tcatcagaga agatatgaaa tataaagatg ctactaataa acacagccat ctgcacagag
     1741 aagataaaca tataacgatt gaggatttat ggaaacgatg gaaaacatca gaagttcata
     1801 attggaccct tgaagacact cttcagtggt tgatagagtt tgttgaacta ccccaatatg
     1861 agaagaattt tagagacaac aatgtcaaag gaacgacact tcccaggata gcagtgcacg
     1921 aaccttcatt tatgatctcc cagttgaaaa tcagtgaccg gagtcacaga caaaaacttc
     1981 agctcaaggc attggatgtg gttttgtttg gacctctaac acgcccacct cataactgga
     2041 tgaaagattt tatcctcaca atttctatag taattggtgt tggaggctgc tggtttgctt
     2101 atacgcagaa taagacatca aaagaacatg ttgcaaaaat gatgaaagat ttagagagct
     2161 tacaaactgc agagcaaagt ctaatggact tacaagagag gcttgaaaag gcacaggaag
     2221 aaaacagaaa tgttgctgta gaaaagcaaa atttagagcg caaaatgatg gatgaaatca
     2281 attatgcaaa ggaggaggct tgtcggctga gagagctaag ggagggagct gaatgtgaat
     2341 tgagtagacg tcagtatgca gaacaggaat tggaacaggt tcgcatggct ctgaaaaagg
     2401 ccgaaaaaga atttgaactg agaagcagtt ggtctgttcc agatgcactt cagaaatggc
     2461 ttcagttaac acatgaagta gaagtgcaat actacaatat taaaagacaa aacgctgaaa
     2521 tgcagctagc tattgctaaa gatgaggcag aaaaaattaa aaagaagaga agcacagtct
     2581 ttgggactct gcacgttgca cacagctcct ccctagatga ggtagaccac aaaattctgg
     2641 aagcaaagaa agctctctct gagttgacaa cttgtttacg agaacgactt tttcgctggc
     2701 aacaaattga gaagatctgt ggctttcaga tagcccataa ctcaggactc cccagcctga
     2761 cctcttccct ttattctgat cacagctggg tggtgatgcc cagagtctcc attccaccct
     2821 atccaattgc tggaggagtt gatgacttag atgaagacac acccccaata gtgtcacaat
     2881 ttcccgggac catggctaaa cctcctggat cattagccag aagcagcagc ctgtgccgtt
     2941 cacgccgcag cattgtgccg tcctcgcctc agcctcagcg agctcagctt gctccacacg
     3001 ccccccaccc gtcacaccct cggcaccctc accacccgca acacacacca cactccttgc
     3061 cttcccctga tccagatatc ctctcagtgt caagttgccc tgcgctttat cgaaatgaag
     3121 tggaggaaga ggccatttac ttctctgctg aaaagcaatg ggaagtgcca gacacagctt
     3181 cagaatgtga ctccttaaat tcttccattg gaaggaaaca gtctcctcct ttaagcctcg
     3241 agatatacca aacattatct ccgcgaaaga tatcaagaga tgaggtgtcc ctagaggatt
     3301 cctcccgagg ggattcgcct gtaactgtgg atgtgtcttg gggttctccc gactgtgtag
     3361 gtctgacaga gactaagagt atgatcttca gtcctgcaag caaagtgtac aatggcattt
     3421 tggagaaatc ctgtagcatg aaccagcttt ccagtggcat cccggtgcct aaacctcgcc
     3481 acacatcatg ttcctcagct ggcaacgaca gtaaaccagt tcaggaagcc ccaagtgttg
     3541 ccagaataag cagcatccca catgaccttt gtcataatgg agagaaaagc aaaaagccat
     3601 caaaaatcaa aagccttttt aagaagaaat ctaagtgaaa cccagctttc ttgtacaaag
     3661 tggttgatcc ggactcagat ctcgagctca agcttcgaat tctgcagtcg acggtaccgc
     3721 gggcccggga tccaccggat ctagataact gatcataatc agccatacca catttgtaga
     3781 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa
     3841 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     3901 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     3961 actcatcaat gtatcttaac gcgtaaattg taagcgttaa tattttgtta aaattcgcgt
     4021 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     4081 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     4141 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     4201 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     4261 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     4321 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     4381 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     4441 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     4501 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     4561 aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg
     4621 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     4681 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     4741 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     4801 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     4861 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     4921 cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     4981 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     5041 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     5101 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc
     5161 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     5221 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     5281 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     5341 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     5401 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     5461 cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg
     5521 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     5581 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     5641 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     5701 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     5761 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     5821 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     5881 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag
     5941 gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa
     6001 aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca
     6061 gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc
     6121 ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt
     6181 cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa
     6241 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     6301 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     6361 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     6421 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     6481 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
     6541 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     6601 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     6661 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     6721 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     6781 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     6841 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     6901 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     6961 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     7021 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcat
//
