LOCUS       sequence-153361-        6826 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Low affinity fluorescent calcium indicator of the GAP (GFP-Aequorin 
            Protein) family targeted to the endoplasmic reticulum.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6826)
  AUTHORS   Navas-Navarro P, Rojo-Ruiz J, Rodriguez-Prados M, Ganfornina MD, 
            Looger LL, Alonso MT, Garcia-Sancho J
  TITLE     GFP-Aequorin Protein Sensor for Ex Vivo and In Vivo Imaging of 
            Ca(2+) Dynamics in High-Ca(2+) Organelles.
  JOURNAL   Cell Chem Biol. 2016 Jun 1. pii: S2451-9456(16)30163-5. doi: 
            10.1016/j.chembiol.2016.05.010.
  PUBMED    27291400
REFERENCE   2  (bases 1 to 6826)
  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 Chem 
            Biol. 2016 Jun 1. pii: S2451-9456(16)30163-5. doi: 
            10.1016/j.chembiol.2016.05.010."
COMMENT     Sequence Label: pcDNA3_erGAP3
FEATURES             Location/Qualifiers
     source          1..6826
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(2378..2396)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2379..2396)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2416..2433)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2422..2646
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2692..3120
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2779..2798)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2989..3010
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3129..3149)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3134..3464
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3315..3450
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3377..3396
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3531..4325
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3585..3604)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4195..4214
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4499..4620
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4536..4555)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4590..4609
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4669..4685)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4669..4685)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4682..4704)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4693..4709
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4717..4723,4724..4741,4742..4747))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4762..4783
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4900..4917)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5071..5659)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5151..5170)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5830..6621,6622..6690))
                     /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     6453..6472
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6691..6795)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctccaaagc
      901 ttccgccacc atgctcctct ctgtgcccct cctcctcggt ctcctgggac tggccgtagc
      961 caatgacgcc tcaaaaggcg aagagctgtt cacaggcgtg gtccctatcc aggtggaact
     1021 cgatggagac gtgaacggcc ataaattctc cgtatcaggc gagggggaag gagacgccac
     1081 atatggaaag ctgacactca aatttatctg caccacagga aaactgcctg tcccatggcc
     1141 aaccctcgtg acaaccttct catacggcgt gcaatgtttt agcagatatc cagaccacat
     1201 gaaaaggcat gacttcttta agagcgcaat gccagaagga tacgtccagg agaggaccat
     1261 tagcttcaag gacgatggca attataagac aagagccgag gtcaaattcg agggagacac
     1321 tctcgtgaac agaattgaac tgaaagggat cgattttaaa gaagatggca acatcctcgg
     1381 acataagctc gagtacaact acaactctca taatgtgtac ataaccgccg acaaacagaa
     1441 gaacgggatc aaagctaatt tcaaggtgag gcacaacata gaagacggcg gcgtacagct
     1501 ggcttaccac tatcagcaga acacacccat aggggacggt cccgtgctcc tgccagataa
     1561 tcactatctg tccacacaat cagccctgag caaggaccct aacgagaaaa gagaccatat
     1621 ggtcctgctg gaatttgtaa cagctgcagg cattactcac ggaatggacg agctgtataa
     1681 gcccggaaca gccactccag ccactacacc caccaccgca cccactgcag gtaccgggaa
     1741 actgaccagc gacttcgata atccaagatg gatagggaga cacaagcata tgttcaattt
     1801 tctgaatgtc aaccacaatg gcaagatatc actggacgaa atggtctaca aggcatctga
     1861 cattgtgatc aacaatctcg gagctactcc tgagcaggca aagaggcaca aagacgccgt
     1921 agaggccttt ttcggtggcg cagggatgaa atacggcgta gagactgatt ggcccgccta
     1981 tattgagggg tggaagaaac tggcaaccga tgaactcgaa aaatacgcta aaaacgagcc
     2041 tactctcata agaatctggg gcgatgctct cttcgatatt gtggataaag cacagaatgg
     2101 agcaataaca ctggatgaat ggaaggcata cactaaggca gctggaatca tccaatcctc
     2161 cgaggattgt gaagagacct tcagggtctg cgacatagac gaatctggcc agctcgacgt
     2221 cgatgagatg acaaggcaac acctgggttt ttggtatacc atggaccctg cttgcgagaa
     2281 actctacggc ggggctgtgc caaaggatga actgtaggaa ttctgcagat atccatcaca
     2341 ctggcggccg ctcgagcatg catctagagg gccctattct atagtgtcac ctaaatgcta
     2401 gagctcgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
     2461 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     2521 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     2581 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct
     2641 ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct
     2701 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     2761 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     2821 gctttccccg tcaagctcta aatcggggca tccctttagg gttccgattt agtgctttac
     2881 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     2941 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     3001 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     3061 tggggatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     3121 aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc caggcaggca
     3181 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     3241 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     3301 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     3361 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc
     3421 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt
     3481 gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac
     3541 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     3601 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     3661 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
     3721 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     3781 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     3841 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     3901 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     3961 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     4021 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc
     4081 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     4141 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     4201 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     4261 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     4321 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     4381 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     4441 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     4501 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     4561 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     4621 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     4681 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     4741 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     4801 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     4861 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     4921 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     4981 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     5041 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     5101 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     5161 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     5221 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag
     5281 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     5341 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     5401 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     5461 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     5521 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     5581 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     5641 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     5701 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     5761 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     5821 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     5881 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     5941 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     6001 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     6061 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     6121 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     6181 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     6241 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     6301 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     6361 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     6421 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     6481 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     6541 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     6601 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     6661 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     6721 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     6781 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
