LOCUS       Exported                6302 bp ds-DNA     circular SYN 18-OCT-2023
DEFINITION  Red fluorescent indicator for calcium signaling in the endoplasmic 
            reticulum.
ACCESSION   .
VERSION     .
KEYWORDS    pCMV R-CEPIA1er
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6302)
  AUTHORS   Suzuki J, Kanemaru K, Ishii K, Ohkura M, Okubo Y, Iino M
  TITLE     Imaging intraorganellar Ca(2+) at subcellular resolution using 
            CEPIA.
  JOURNAL   Nat Commun. 2014 Jun 13;5:4153. doi: 10.1038/ncomms5153.
  PUBMED    24923787
REFERENCE   2  (bases 1 to 6302)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 18, 2023 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6302
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        45..421
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        423..626
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     577..597
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      683..692
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     sig_peptide     join(689..733,816..827)
                     /product="modified signal sequence from the mouse 
                     immunoglobulin heavy chain variable region"
                     /label=IgH signal sequence
     CDS             864..920
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKAGHAVRAIGRLSS"
     primer_bind     1616..1635
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1659..2099
                     /codon_start=1
                     /product="calcium-binding protein"
                     /label=calmodulin
                     /translation="DQLTEEQIAEFKEAFSLFDKDGDGTITTKDLGTVLRSLGQNPTEA
                     ELQDMINEVDADGNGTIDFPDFLTMMARKMKDTDSEEEIREAFRVWDKDGNGYISAADL
                     RHVMTNLGEKLTDEEVDEMIREADIDGEGQVNYEEFVQMMTAK"
     CDS             2109..2138
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(2192..2209)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2198..2422
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2468..2896
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2555..2574)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2765..2786
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2905..2925)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2910..3239
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3090..3225
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3152..3171
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3265..4059
                     /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="MVEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3319..3338)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3929..3948
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4233..4354
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4270..4289)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4324..4343
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4387..4404)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4558..5146)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4638..4657)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5317..6177)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5940..5959
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6178..6282)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cacctgacgt cagatcgacg gatcgggaga tcggtaccga attcacattg attattgagt
       61 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
      121 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
      181 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
      241 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
      301 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
      361 atgaccttat gggactttcc tacttggcag tacatctacg gttagtcatc gctattacca
      421 tagtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
      481 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      541 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
      601 ggtgggaggt ctatataagc agagctttct ggctaactag agaacccact gcttactggc
      661 acgtggaaat taatacgacg tggccaccat gggatggagc tgtatcatcc tcttcttggt
      721 agcaacagct acaggtaagg ggttaacagt agcaggcttg aggtctggac atatatatgg
      781 gtgacaatga catccacttt gcctttctct ccacaggcgc gcactcccag gtccaactgc
      841 agggatccat ggtcgactct tcacgtcgta agtggaataa ggcaggtcac gcagtcagag
      901 ctataggtcg gctgagctca cccgtggttt ccgagcggat gtaccccgag gacggcgccc
      961 tgaagagcga gatcaagaag gggctgaggc tgaaggacgg cggccactac gccgccgagg
     1021 tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac atcgtcgaca
     1081 tcaagttgga catcgtgtcc cacaacgagg actacaccat cgtggaacag tgcgaacgcg
     1141 ccgagggccg ccactccacc ggcggcatgg acgagctgta caagggaggt acaggcggga
     1201 gtctggtgag caagggcgag gaggataaca tggccatcat caaggagttc atgcgcttca
     1261 aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc gagggcgagg
     1321 gccgccccta cgaggccttt cagaccgcta agctgaaggt gaccaagggt ggccccctgc
     1381 ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc tacattaagc
     1441 acccagccga catccccgac tacttcaagc tgtccttccc cgagggcttc aggtgggagc
     1501 gcgtgatgaa cttcgaggac ggcggcatta ttcacgttaa ccaggactcc tccctgcagg
     1561 acggcgtatt catctacaag gtgaagctgc gcggcaccaa cttccccccc gacggccccg
     1621 taatgcagaa gaagaccatg ggctgggagg ctactcgaga ccaactgact gaagagcaga
     1681 tcgcagaatt taaagaggct ttctccctat ttgacaagga cggggatggg acaataacaa
     1741 ccaaggatct ggggacggtg ctgcggtctc tggggcagaa ccccacagaa gcagagctcc
     1801 aggacatgat caatgaagta gatgccgacg gtaatggcac aatcgacttc cctgatttcc
     1861 tgacaatgat ggcaagaaaa atgaaagaca cagacagtga agaagaaatt cgcgaagcgt
     1921 tccgtgtgtg ggataaggat ggcaatggct acatctctgc agcagacctt cgccacgtga
     1981 tgacaaacct tggagagaag ttaacagatg aagaggttga tgaaatgatc agggaagcag
     2041 atatcgatgg agaaggtcag gtaaactacg aagagtttgt acaaatgatg acagcgaagg
     2101 cggccgcaga acaaaaactc atctcagaag aggatctgaa tggggccgca agcgagaagg
     2161 acgagctgta gtctagaagc tcgctgatca gcctcgactg tgccttctag ttgccagcca
     2221 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     2281 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     2341 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     2401 ggggatggcc cgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg
     2461 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     2521 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     2581 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     2641 cgatttagtg ctttacggca cctctccccc aaaaaacttg attagggtga tggttcacgt
     2701 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     2761 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     2821 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     2881 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     2941 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     3001 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3061 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     3121 attctccgcc cctaggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
     3181 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     3241 agctcccccc ccgggaggtc cacaatggtt gaacaagatg gattgcacgc aggttctccg
     3301 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     3361 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     3421 ctgtccggtg ccctgaatga actccaggac gaggcagcgc ggctatcgtg gctggccacg
     3481 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     3541 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     3601 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     3661 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     3721 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     3781 aggctcaagg cgcgtatgcc cgacggcgag gatctcgtcg tgactcatgg cgatgcctgc
     3841 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     3901 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     3961 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     4021 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     4081 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     4141 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     4201 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     4261 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     4321 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgatctttc
     4381 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     4441 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     4501 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     4561 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     4621 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     4681 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     4741 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     4801 gctgggctgt atgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     4861 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     4921 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     4981 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     5041 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     5101 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     5161 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     5221 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     5281 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     5341 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     5401 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     5461 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     5521 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     5581 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     5641 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     5701 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     5761 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     5821 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     5881 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     5941 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     6001 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgggc
     6061 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     6121 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     6181 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     6241 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     6301 gc
//
