LOCUS       SK0537(pCMV-EGFP        6039 bp    DNA     circular SYN 03-AUG-2021
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6039)
  AUTHORS   Institute for Basic Science
  TITLE     Direct Submission
  JOURNAL   Exported Oct 22, 2025 from SnapGene 8.1.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6039
                     /mol_type="other DNA"
                     /note="color: #ffffff"
                     /organism="recombinant plasmid"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
                     /note="color: #ffffff"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /note="color: #ffffff; direction: RIGHT"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /note="This feature has 3 segments:
                        1:  613 ..  615 / #05fd14
                        2:  616 ..  618 / #05fd14 / 1a
                        3:  619 .. 1329 / #05fd14"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     misc_feature    1330
                     /label=MCS
                     /note="multiple cloning site of fluorescent protein
                     plasmids"
                     /note="color: #99ccff"
     CDS             1345..2688
                     /codon_start=1
                     /label=STIM1(238-685)
                     /note="color: #ff0000"
                     /translation="KEHMKKMMKDLEGLHRAEQSLHDLQERLHKAQEEHRTVEVEKVHL
                     EKKLRDEINLAKQEAQRLKELREGTENERSRQKYAEEELEQVREALRKAEKELESHSSW
                     YAPEALQKWLQLTHEVEVQYYNIKKQNAEKQLLVAKEGAEKIKKKRNTLFGTFHVAHSS
                     SLDDVDHKILTAKQALSEVTAALRERLHRWQQIEILCGFQIVNNPGIHSLVAALNIDPS
                     WMGSTRPNPAHFIMTDDVDDMDEEIVSPLSMQSPSLQSSVRQRLTEPQHGLGSQRDLTH
                     SDSESSLHMSDRQRVAPKPPQMSRAADEALNAMTSNGSHRLIEGVHPGSLVEKLPDSPA
                     LAKKALLALNHGLDKAHSLMELSPSAPPGGSPHLDSSRSHSPSSPDPDTPSPVGDSRAL
                     QASRNTRIPHLAGKKAVAEEDNGSIGEETDSSPGRKKFPLKIFKKPLKK"
     primer_bind     1585..1607
                     /label=BspEI-STIM1(318~)-JGSF
                     /note="color: black; sequence: 
                     GCGGAGGGGGAGGCTCCGGAGAGGAGGAGTTGGAGCAGGTTCG"
     primer_bind     complement(1979..1995)
                     /label=STIM1(~454)-stop-BamHI-JGSR
                     /note="color: black; sequence: 
                     TCAGTTATCTAGATCCGGTGGATCCCTAGAGGGCAGCCACCAGTG"
     primer_bind     complement(1984..2004)
                     /label=STIM1(~457)-stop-BamHI-JGSR
                     /note="color: black; sequence: 
                     TCAGTTATCTAGATCCGGTGGATCCCTAGTCTATGTTGAGGGCAGCCAC"
     primer_bind     complement(1993..2013)
                     /label=STIM1(~460)-stop-BamHI-JGSR
                     /note="color: black; sequence: 
                     TCAGTTATCTAGATCCGGTGGATCCCTACCAGCTGGGGTCTATGTTGAG"
     primer_bind     complement(2005..2022)
                     /label=STIM1(~463)-BamHI-JGSR
                     /note="color: black; sequence: 
                     CAGTTATCTAGATCCGGTGGATCCCTAACTGCCCATCCAGCTGGG"
     primer_bind     complement(2697..2722)
                     /label=STIM1(~454)-stop-BamHI-JGSR
                     /note="color: black; sequence: 
                     TCAGTTATCTAGATCCGGTGGATCCCTAGAGGGCAGCCACCAGTG"
     primer_bind     complement(2697..2722)
                     /label=STIM1(~457)-stop-BamHI-JGSR
                     /note="color: black; sequence: 
                     TCAGTTATCTAGATCCGGTGGATCCCTAGTCTATGTTGAGGGCAGCCAC"
     primer_bind     complement(2697..2722)
                     /label=STIM1(~460)-stop-BamHI-JGSR
                     /note="color: black; sequence: 
                     TCAGTTATCTAGATCCGGTGGATCCCTACCAGCTGGGGTCTATGTTGAG"
     primer_bind     complement(2697..2721)
                     /label=STIM1(~463)-BamHI-JGSR
                     /note="color: black; sequence: 
                     CAGTTATCTAGATCCGGTGGATCCCTAACTGCCCATCCAGCTGGG"
     misc_feature    2699..2703
                     /label=MCS
                     /note="multiple cloning site of fluorescent protein
                     plasmids"
                     /note="color: #99ccff"
     misc_feature    2712..2722
                     /label=stop codons
                     /note="stop codons in all three reading frames"
                     /note="color: #ff0000"
     polyA_signal    2827..2948
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
                     /note="color: #a6acb3"
     rep_origin      complement(2955..3410)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     promoter        3437..3541
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: RIGHT"
     promoter        3543..3900
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
                     /note="color: #ffffff; direction: RIGHT"
     rep_origin      3751..3886
                     /label=SV40 ori
                     /note="SV40 origin of replication"
                     /note="color: #ffff00"
     CDS             3935..4729
                     /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)"
                     /note="color: #ccffcc"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     misc_feature    4961..5008
                     /label=HSV TK poly(A) signal
                     /note="herpesvirus thymidine kinase polyadenylation signal"
                     /note="color: #a6acb3"
     rep_origin      5337..5925
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
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 gctagcgcta
      601 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1321 ctgtacaagt ccggactcag atctaaggag cacatgaaga agatgatgaa ggacttggag
     1381 gggttacacc gagctgagca gagtctgcat gaccttcagg aaaggctgca caaggcccag
     1441 gaggagcacc gcacagtgga ggtggagaag gtccatctgg aaaagaagct gcgcgatgag
     1501 atcaaccttg ctaagcagga agcccagcgg ctgaaggagc tgcgggaggg tactgagaat
     1561 gagcggagcc gccaaaaata tgctgaggag gagttggagc aggttcggga ggccttgagg
     1621 aaagcagaga aggagctaga atctcacagc tcatggtatg ctccagaggc ccttcagaag
     1681 tggctgcagc tgacacatga ggtggaggtg caatattaca acatcaagaa gcaaaatgct
     1741 gagaagcagc tgctggtggc caaggagggg gctgagaaga taaaaaagaa gagaaacaca
     1801 ctctttggca ccttccacgt ggcccacagc tcttccctgg atgatgtaga tcataaaatt
     1861 ctaacagcta agcaagcact gagcgaggtg acagcagcat tgcgggagcg cctgcaccgc
     1921 tggcaacaga tcgagatcct ctgtggcttc cagattgtca acaaccctgg catccactca
     1981 ctggtggctg ccctcaacat agaccccagc tggatgggca gtacacgccc caaccctgct
     2041 cacttcatca tgactgacga cgtggatgac atggatgagg agattgtgtc tcccttgtcc
     2101 atgcagtccc ctagcctgca gagcagtgtt cggcagcgcc tgacggagcc acagcatggc
     2161 ctgggatctc agagggattt gacccattcc gattcggagt cctccctcca catgagtgac
     2221 cgccagcgtg tggcccccaa acctcctcag atgagccgtg ctgcagacga ggctctcaat
     2281 gccatgactt ccaatggcag ccaccggctg atcgaggggg tccacccagg gtctctggtg
     2341 gagaaactgc ctgacagccc tgccctggcc aagaaggcat tactggcgct gaaccatggg
     2401 ctggacaagg cccacagcct gatggagctg agcccctcag ccccacctgg tggctctcca
     2461 catttggatt cttcccgttc tcacagcccc agctccccag acccagacac accatctcca
     2521 gttggggaca gccgagccct gcaagccagc cgaaacacac gcattcccca cctggctggc
     2581 aagaaggctg tggctgagga ggataatggc tctattggcg aggaaacaga ctccagccca
     2641 ggccggaaga agtttcctct caaaatcttt aagaagcctc ttaagaagta ggaattggga
     2701 tccaccggat ctagataact gatcataatc agccatacca catttgtaga ggttttactt
     2761 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt
     2821 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     2881 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     2941 gtatcttaac gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
     3001 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     3061 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     3121 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     3181 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     3241 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     3301 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
     3361 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac
     3421 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     3481 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     3541 tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     3601 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     3661 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3721 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     3781 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
     3841 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     3901 gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc
     3961 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     4021 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     4081 caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg
     4141 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     4201 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     4261 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     4321 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     4381 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     4441 actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg
     4501 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     4561 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     4621 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     4681 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     4741 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     4801 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     4861 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa
     4921 acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat
     4981 aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac
     5041 tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc
     5101 ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca
     5161 ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt
     5221 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     5281 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     5341 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     5401 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     5461 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     5521 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     5581 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     5641 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     5701 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     5761 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     5821 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     5881 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     5941 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     6001 tcccctgatt ctgtggataa ccgtattacc gccatgcat
//
