LOCUS       sequence-315099-        7336 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Neuronal expression of scFLARE2 component.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7336)
  AUTHORS   Sanchez MI, Nguyen QA, Wang W, Soltesz I, Ting AY
  TITLE     Transcriptional readout of neuronal activity via an engineered 
            Ca(2+)-activated protease.
  JOURNAL   Proc Natl Acad Sci U S A. 2020 Dec 15. pii: 2006521117. doi: 
            10.1073/pnas.2006521117.
  PUBMED    33323488
REFERENCE   2  (bases 1 to 7336)
  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: "Proc Natl 
            Acad Sci U S A. 2020 Dec 15. pii: 2006521117. doi: 
            10.1073/pnas.2006521117."
COMMENT     Sequence Label: scFLARE2
FEATURES             Location/Qualifiers
     source          1..7336
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..448
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             631..657
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             883..1392
                     /codon_start=1
                     /product="N-terminal portion of the CIB1 transcription 
                     factor from Arabidopsis thaliana"
                     /label=CIBN
                     /note="binds to CRY2 (cryptochrome 2) that has absorbed 
                     blue light (Kennedy et al., 2010)"
                     /translation="MNGAIGGDLLLNFPDMSVLERQRAHLKYLNPTFDSPLAGFFADSS
                     MITGGEMDSYLSTAGLNLPMMYGETTVEGDSRLSISPETTLGTGNFKAAKFDTETKDCN
                     EAAKKMTMNRDDLVEEGEEEKSKITEQNNGSTKSIKKMKHKAKKEENNFSNDSSKVTKE
                     LEKTDYI"
     CDS             3079..3099
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQM"
     CDS             3112..3135
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             join(3136..3756,3757..4143)
                     /codon_start=1
                     /product="tetracycline-controlled transactivator, 
                     comprising a fusion of the tetracycline repressor TetR with
                     the C-terminal activation domain of herpes simplex virus 
                     VP16"
                     /label=tTA
                     /note="In the Tet-Off(R) system, tTA binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the absence of tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGSAYSRARTKNNYGSTI
                     EGLLDLPDDDAPEEAGLAAPRLSFLPAGHTRRLSTAPPTDVSLGDELHLDGEDVAMAHA
                     DALDDFDLDMLGDGDSPGPGFTPHDSAPYGALDMADFEFEQMFTDALGIDEYGG"
     primer_bind     complement(3209..3228)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     primer_bind     complement(4177..4194)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4183..4390
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4419..4559
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4419..4548
                     /label=AAV2 ITR
     rep_origin      4634..5089
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4721..4740)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4931..4952
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5106..5125)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5225..5247
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5285..5303)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5371..5475
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5476..5544,5545..6336)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEVDKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5694..5713)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6507..7095
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6996..7015
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   7157..7286
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 agtgcaagtg ggttttagga ccaggatgag gcggggtggg ggtgcctacc tgacgaccga
       61 ccccgaccca ctggacaagc acccaacccc cattccccaa attgcgcatc ccctatcaga
      121 gagggggagg ggaaacagga tgcggcgagg cgcgtgcgca ctgccagctt cagcaccgcg
      181 gacagtgcct tcgcccccgc ctggcggcgc gcgccaccgc cgcctcagca ctgaaggcgc
      241 gctgacgtca ctcgccggtc ccccgcaaac tccccttccc ggccaccttg gtcgcgtccg
      301 cgccgccgcc ggcccagccg gaccgcacca cgcgaggcgc gagatagggg ggcacgggcg
      361 cgaccatctg cgctgcggcg ccggcgactc agcgctgcct cagtctgcgg tgggcagcgg
      421 aggagtcgtg tcgtgcctga gagcgcagtc gagaagaagc ttgccaccat gcacctgaga
      481 atccacgcga gacggagccc tcctcgccgg ccggcctgga cgcttgggat ctggttcctg
      541 ttctggggat gtatcgtcag ctctgtatgg agtcaattgt cttctaatgt agcttcctcc
      601 tcctccacct cttcctcgcc ggggtctcac tacccatacg atgtgccaga ttacgccaac
      661 cccacggagc cgggaatcag acgggttccg ggggcctcag aggtgatccg ggagtcgagc
      721 agcacaacag ggatggtcgt cggcattgtg gctgctgccg ccctctgcat cttgatcctc
      781 ctgtacgcca tgtacaagta caggaacagg gacgagggct cgggctcgac ctcgggctcg
      841 ggcagcggtg gctctggagg ttctggtggc agctctggag gtatgaatgg agctatagga
      901 ggtgaccttt tgctcaattt tcctgacatg tcggtcctag agcgccaaag ggctcacctc
      961 aagtacctca atcccacctt tgattctcct ctcgccggct tctttgccga ttcttcaatg
     1021 attaccggcg gcgagatgga cagctatctt tcgactgccg gtttgaatct tccgatgatg
     1081 tacggtgaga cgacggtgga aggtgattca agactctcaa tttcgccgga aacgacgctt
     1141 gggactggaa atttcaaggc agcgaagttt gatacagaga ctaaggattg taatgaggcg
     1201 gcgaagaaga tgacgatgaa cagagatgac ctagtagaag aaggagaaga agagaagtcg
     1261 aaaataacag agcaaaacaa tgggagcaca aaaagcatca agaagatgaa acacaaagcc
     1321 aagaaagaag agaacaattt ctctaatgat tcatctaaag tgacgaagga attggagaaa
     1381 acggattata ttcatggtgg ctctggtagc ggagaaagct tgtttaaggg accacgtgat
     1441 tacaacccga tatcgagcac catttgtcat ttgacgaatg aatctgatgg gcacacaaca
     1501 tcgttgtatg gtattggatt tggtcccttc atcattacaa acaagcactt gtttagaaga
     1561 aataatggaa cactgttggt ccaatcacta catctcgagg accaactgac tgaagagcag
     1621 atcgcagaat ttaaagagct cttctcccta tttgacaagg acggggatgg gacaataaca
     1681 accaaggagc tggggacggt gatgcggtct ctggggcaga accccacaga agcagagctg
     1741 caagacatga tcaatgaagt agatgccgac ggtgacggca caatcgactt ccctgagttc
     1801 ctgaccatga tggctagaaa aatgaaggac acaggcggtg tgaagctgat cccctcctgg
     1861 accaccgtga tcctggtgaa gagcatgctg cgcaagcgca gcttcggcaa ccccttcggt
     1921 ggcgacagcg aagaggaaat ccgagaagca ttcggtgtgt ttgataagga tggcaatggc
     1981 tacatcagtg cagcagagct tcgccacgtg atgacaaacc ttggagagaa gttaacagat
     2041 gaagaggttg atgaaatgat cagggaagca gacatcgatg gggatggtca ggtaaactac
     2101 gaagagtttg tacaaatgat gacagcgaag gcggccgcgg gtgtattcaa ggtcaagaac
     2161 accacgactt tgcaacaaca cctcattgat gggagggaca tgataattat tcgcatgcct
     2221 aaggatttcc caccatttcc tcaaaagctg aaatttagag agccacaaag ggaagagcgc
     2281 atatgtcttg tgacaaccaa cttccaaact aagagcatgt ctagcatggt gtcagacact
     2341 agttgcacat tcccttcatc tgatggcata ttctggaagc attggattca aaccaaggat
     2401 gggcagtgtg gcaatccatt agtatcaact agagatgggt tcattgttgg tatacactca
     2461 gcatcgaatt tcaccaacac aaacaattat ttcacaagcg tgccgaaaaa cttcatggaa
     2521 ttgttgacaa atcaggaggc gcagcagtgg gttagtggtt ggcgattaaa tgctgactca
     2581 gtattgtggg gtggccataa agttttcatg gtggagctgg ccgagaagct ggccggcctg
     2641 gacatcaacg gtggagctag cgagtttcgg gcaaccacac tggaacggat cgagaaaagc
     2701 ttcgtgatta ctgatccgag actgcctgac aacccaatca tttttgtgag cgattccttc
     2761 ctgcagctga cagaatattc tcgggaagag atcctggggc gcaattgccg ttttctgcag
     2821 ggacccgaga cagaccgtgc cactgttcgg aaaatcagag atgctattga caaccagact
     2881 gaagtgaccg ttcagctgat caattatacc aagagcggca agaagttctg gaacgtgttc
     2941 cacctgcagc cgatgcgcga ttataagggc gacgtccagt acttcattgg cgtgcagctg
     3001 gatggcaccg aacgtcttca tggcgccgct gagcgtgagg cggtctgcct ggtcaaaaag
     3061 acagcctttc agattgctga gaacctgtac ttccagatgg gcggaggatc cgactacaag
     3121 gatgacgacg ataagatgtc tagattagat aaaagtaaag tgattaacag cgcattagag
     3181 ctgcttaatg aggtcggaat cgaaggttta acaacccgta aactcgccca gaagctaggt
     3241 gtagagcagc ctacattgta ttggcatgta aaaaataagc gggctttgct cgacgcctta
     3301 gccattgaga tgttagatag gcaccatact cacttttgcc ctttagaagg ggaaagctgg
     3361 caagattttt tacgtaataa cgctaaaagt tttagatgtg ctttactaag tcatcgcgat
     3421 ggagcaaaag tacatttagg tacacggcct acagaaaaac agtatgaaac tctcgaaaat
     3481 caattagcct ttttatgcca acaaggtttt tcactagaga atgcattata tgcactcagc
     3541 gctgtggggc attttacttt aggttgcgta ttggaagatc aagagcatca agtcgctaaa
     3601 gaagaaaggg aaacacctac tactgatagt atgccgccat tattacgaca agctatcgaa
     3661 ttatttgatc accaaggtgc agagccagcc ttcttattcg gccttgaatt gatcatatgc
     3721 ggattagaaa aacaacttaa atgtgaaagt gggtccgcgt acagccgcgc gcgtacgaaa
     3781 aacaattacg ggtctaccat cgagggcctg ctcgatctcc cggacgacga cgcccccgaa
     3841 gaggcggggc tggcggctcc gcgcctgtcc tttctccccg cgggacacac gcgcagactg
     3901 tcgacggccc ccccgaccga tgtcagcctg ggggacgagc tccacttaga cggcgaggac
     3961 gtggcgatgg cgcatgccga cgcgctagac gatttcgatc tggacatgtt gggggacggg
     4021 gattccccgg gtccgggatt taccccccac gactccgccc cctacggcgc tctggatatg
     4081 gccgacttcg agtttgagca gatgtttacc gatgcccttg gaattgacga gtacggtggg
     4141 taataggaat tcctcgagct agagctcgct gatcagcctc gactgtgcct tctagttgcc
     4201 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     4261 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     4321 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     4381 atgctgggga ggtaaccacg tgcggaccga gcggccgcag gaacccctag tgatggagtt
     4441 ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg
     4501 acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg
     4561 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca
     4621 aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     4681 cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct
     4741 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     4801 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt
     4861 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     4921 ttctttaata gtggactctt gttccaaact ggaacaacac tcaactctat ctcgggctat
     4981 tcttttgatt tataagggat tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt
     5041 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact
     5101 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc
     5161 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     5221 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga
     5281 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     5341 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     5401 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     5461 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     5521 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     5581 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     5641 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     5701 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     5761 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     5821 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     5881 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     5941 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     6001 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     6061 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggtgga taaagttgca
     6121 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     6181 ggtgagcgtg gaagccgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     6241 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     6301 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     6361 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     6421 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     6481 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     6541 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     6601 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     6661 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     6721 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     6781 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     6841 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     6901 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     6961 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     7021 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     7081 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     7141 ccttttgctc acatgtcctg caggcagctg cgcgctcgct cgctcactga ggccgcccgg
     7201 gcgtcgggcg acctttggtc gcccggcctc agtgagcgag cgagcgcgca gagagggagt
     7261 ggccaactcc atcactaggg gttcctgcgg ccgcacgcgt tgtacagtgt ctagactgca
     7321 gagggccctg cgtatg
//
