LOCUS       sequence-264411-        7382 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses IP3KC GFP tagged.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7382)
  AUTHORS   Dewaste V, Moreau C, De Smedt F, Bex F, De Smedt H, Wuytack F, 
            Missiaen L, Erneux C
  TITLE     The three isoenzymes of human inositol-1,4,5-trisphosphate 3-kinase 
            show specific intracellular localization but comparable Ca2+ 
            responses on transfection in COS-7 cells.
  JOURNAL   Biochem J. 2003 Aug 15;374(Pt 1):41-9.
  PUBMED    12747803
REFERENCE   2  (bases 1 to 7382)
  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: "Biochem 
            J."; date: "2003-08-15"; volume: "374(Pt 1)"; pages: "41-9"
COMMENT     Sequence Label: GFP-IP3KC
FEATURES             Location/Qualifiers
     source          1..7382
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        2..305
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        306..509
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     460..480
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      548..557
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(554..556,557..559,560..1270)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(554..556,557..559,560..1270)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(599..620)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(860..879)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1207..1228
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4111..4232
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4148..4167)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4202..4221
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4239..4694)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4376..4397)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4588..4607
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4721..4825
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4827..5184
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4827..5018
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4850..4870)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5035..5170
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5219..6013
                     /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             5219..6013
                     /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(5273..5292)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5883..5902
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6201..6220)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6245..6292
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6621..7209
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7110..7129
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
       61 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
      121 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
      181 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
      241 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
      301 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
      361 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
      421 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
      481 tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgctagcgct
      541 accggtcgcc accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
      601 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg
      661 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
      721 gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc
      781 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
      841 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
      901 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
      961 catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga
     1021 caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag
     1081 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     1141 gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg
     1201 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
     1261 gctgtacaag tccggactca gatccggcat gaggcgctgc ccgtgccgtg ggagcctgaa
     1321 cgaggcggag gccggggcgc tgcccgcggc ggcccgcatg ggactggagg cgccgcgagg
     1381 agggcggcgg cggcagccgg gacagcagcg acctgggccc ggcgcagggg ccccggcggg
     1441 gcggccggag gggggcgggc cctgggcccg gacagagggg tccagcctcc acagcgagcc
     1501 tgagagggcc ggcctcgggc ctgcgccggg gacagagagt ccgcaggcag aattctggac
     1561 agacggacag actgagcccg cggcagctgg ccttggagta gagaccgaga ggcccaagca
     1621 aaagacggag ccagacaggt ccagcctccg gacgcatcta gaatggagct ggtcagagct
     1681 ggagacgact tgtctttgga cggagaccgg gacagatggc ctttggactg atccgcacag
     1741 gtccgacctc cagtttcagc ccgaggaggc cagcccctgg acacagccag gggttcatgg
     1801 gccctggaca gagctggaaa cgcatgggtc acagactcag ccagagaggg tcaagtcctg
     1861 ggctgataac ctctggaccc accagaacag ttccagcctc cagactcacc cagaaggagc
     1921 ctgtccctca aaagagccaa gtgctgatgg ctcctggaaa gaattgtata ctgatggctc
     1981 caggacacaa caggatattg aaggtccctg gacagagcca tatactgatg gctcccagaa
     2041 aaaacaggat actgaagcag ccaggaaaca gcctggcact ggtggtttcc aaatacaaca
     2101 ggatactgat ggctcctgga cacaacctag cactgacggt tcccagacag cacctgggac
     2161 agactgcctc ttgggagagc ctgaggatgg cccattagag gaaccagagc ctggagaatt
     2221 gctgactcac ctgtactctc acctgaagtg tagccccctg tgccctgtgc cccgcctcat
     2281 cattacccct gagacccctg agcctgaggc ccagccagtg ggacccccct cccgggttga
     2341 ggggggcagc ggcggcttct cctctgcctc ttctttcgac gagtctgagg atgacgtggt
     2401 ggccgggggc ggaggtgcca gcgatcccga ggacaggtct gggagcaaac cctggaagaa
     2461 gctgaagaca gttctgaagt attcaccctt tgtggtctcc ttccgaaaac actacccttg
     2521 ggtccagctt tctggacatg ctgggaactt ccaggcagga gaggatggtc ggattctgaa
     2581 acgtttctgt cagtgtgagc agcgcagcct ggagcagctg atgaaagacc cgctgcgacc
     2641 tttcgtgcct gcctactatg gcatggtgct gcaggatggc cagaccttca accagatgga
     2701 agacctcctg gctgactttg agggcccctc cattatggac tgcaagatgg gcagcaggac
     2761 ctatctggaa gaggagctag tgaaggcacg ggaacgtccc cgtccccgga aggacatgta
     2821 tgagaagatg gtggctgtgg accctggggc ccctacccct gaggagcatg cccagggtgc
     2881 agtcaccaag ccccgctaca tgcagtggag ggaaaccatg agctccacct ctaccctggg
     2941 cttccggatc gagggcatca agaaggcaga tgggacctgt aacaccaact tcaagaagac
     3001 gcaggcactg gagcaggtga caaaagtgct ggaggacttc gtggatggag accacgtcat
     3061 cctgcaaaag tacgtggcat gcctagaaga acttcgtgaa gctctggaga tctccccctt
     3121 cttcaagacc cacgaggtgg taggcagctc cctcctcttc gtgcacgacc acaccggcct
     3181 ggccaaggtc tggatgatag acttcggcaa gacggtggcc ttgcccgacc accagacgct
     3241 cagccacagg ctgccctggg ctgagggcaa ccgtgaggac ggctacctct ggggcctgga
     3301 caacatgatc tgcctcctgc aggggctggc acagagctga gctgctcagc caccatcagg
     3361 ttaattggat ggcgccagtc tggctggagg agccctgaga tgccatggga ggcctgaggt
     3421 tggccacggg ggagctggcc tccagggacg ggagagattg tgtcatgtgc cacacgagac
     3481 caacgtggaa aagtctgaag ggccttggga gaccaggtag cacctggccc catcatgatg
     3541 caggggtttt ggggacctgg aaggaaggtg atgaggcagt gagtcagaaa aaccagaacg
     3601 gggtccccgg atctgccggg aaggcttctg aggggctgcc ctgagagcat tcagttcaca
     3661 tgtcacaggg tatggtgtga cagggtgcct gtggacacat gaatcacttc taacctgcct
     3721 ccctgtcagc ctccaggctg ccagctggct gaggccaggg actgggtcag gctcatctgt
     3781 ggcgcctcag agggtcagca tcattggtga acagatgcag gcgctgctgg accatctggg
     3841 gagagtgaca gtccatgtct tcaccaggga gccatttgag tgctgagcga caagaggctc
     3901 agagggcatg accccatggg actggatgcg gcctgagggc tgataccgct gggcggtatc
     3961 ctgccctgct gtggccctgt gggatccacc ggatctagat aactgatcat aatcagccat
     4021 accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg
     4081 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac
     4141 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     4201 tgtggtttgt ccaaactcat caatgtatct taacgcgtaa attgtaagcg ttaatatttt
     4261 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     4321 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     4381 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     4441 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     4501 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     4561 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     4621 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     4681 gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     4741 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     4801 tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt
     4861 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     4921 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     4981 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     5041 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     5101 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     5161 tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat
     5221 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     5281 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     5341 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     5401 agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     5461 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     5521 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     5581 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     5641 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     5701 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg
     5761 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     5821 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     5881 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     5941 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     6001 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
     6061 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     6121 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     6181 caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct
     6241 atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc
     6301 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa
     6361 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg
     6421 ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt
     6481 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     6541 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     6601 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     6661 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     6721 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     6781 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     6841 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     6901 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     6961 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     7021 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     7081 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     7141 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     7201 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     7261 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg
     7321 cattagttat taatagtaat caattacggg gtcattagtt catagcccat atatggagtt
     7381 cc
//
