LOCUS       sequence-181304-        6375 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transcription factor component of FLARE.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6375)
  AUTHORS   Wang W, Wildes CP, Pattarabanjird T, Sanchez MI, Glober GF, Matthews
            GA, Tye KM, Ting AY
  TITLE     A light- and calcium-gated transcription factor for imaging and 
            manipulating activated neurons.
  JOURNAL   Nat Biotechnol. 2017 Jun 26. doi: 10.1038/nbt.3909.
  PUBMED    28650461
REFERENCE   2  (bases 1 to 6375)
  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: "Nat 
            Biotechnol. 2017 Jun 26. doi: 10.1038/nbt.3909."
COMMENT     Sequence Label: Ca-FLARE (TF)
FEATURES             Location/Qualifiers
     source          1..6375
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             422..448
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             674..1183
                     /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             1910..1930
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQM"
     CDS             1943..1966
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             join(1967..2587,2588..2974)
                     /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(2040..2059)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     primer_bind     complement(3008..3025)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3014..3221
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3250..3390
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3250..3379
                     /label=AAV2 ITR
     rep_origin      3465..3920
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3552..3571)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3762..3783
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3937..3956)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4056..4078
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4116..4134)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4202..4306
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4307..4375,4376..5167)
                     /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(4525..4544)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5338..5926
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5827..5846
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   5988..6117
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        join(6168..6375,1..240)
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
ORIGIN
        1 gcgcgccacc gccgcctcag cactgaaggc gcgctgacgt cactcgccgg tcccccgcaa
       61 actccccttc ccggccacct tggtcgcgtc cgcgccgccg ccggcccagc cggaccgcac
      121 cacgcgaggc gcgagatagg ggggcacggg cgcgaccatc tgcgctgcgg cgccggcgac
      181 tcagcgctgc ctcagtctgc ggtgggcagc ggaggagtcg tgtcgtgcct gagagcgcag
      241 tcgagaagga tccgccacca tgcacctgag aatccacgcg agacggagcc ctcctcgccg
      301 gccggcctgg acgcttggga tctggttcct gttctgggga tgtatcgtca gctctgtatg
      361 gagtcaattg tcttctaatg tagcttcctc ctcctccacc tcttcctcgc cggggtctca
      421 ctacccatac gatgtgccag attacgccaa ccccacggag ccgggaatca gacgggttcc
      481 gggggcctca gaggtgatcc gggagtcgag cagcacaaca gggatggtcg tcggcattgt
      541 ggctgctgcc gccctctgca tcttgatcct cctgtacgcc atgtacaagt acaggaacag
      601 ggacgagggc tcgggctcga cctcgggctc gggcagcggt ggctctggag gttctggtgg
      661 cagctctgga ggtatgaatg gagctatagg aggtgacctt ttgctcaatt ttcctgacat
      721 gtcggtccta gagcgccaaa gggctcacct caagtacctc aatcccacct ttgattctcc
      781 tctcgccggc ttctttgccg attcttcaat gattaccggc ggcgagatgg acagctatct
      841 ttcgactgcc ggtttgaatc ttccgatgat gtacggtgag acgacggtgg aaggtgattc
      901 aagactctca atttcgccgg aaacgacgct tgggactgga aatttcaagg cagcgaagtt
      961 tgatacagag actaaggatt gtaatgaggc ggcgaagaag atgacgatga acagagatga
     1021 cctagtagaa gaaggagaag aagagaagtc gaaaataaca gagcaaaaca atgggagcac
     1081 aaaaagcatc aagaagatga aacacaaagc caagaaagaa gagaacaatt tctctaatga
     1141 ttcatctaaa gtgacgaagg aattggagaa aacggattat attcatagcg gtagtggcac
     1201 cgtgcgcgtg ccgattgcgg tgggcgaaag cgattttgaa aacctgaaca ccgaagatgt
     1261 gagcagcgaa agcgatccgg gtggctctgg tagcttcaac gctcgcagga agctggctgg
     1321 tgctatcctg ttcaccatgc tcgccacaag aaacttcagc ggcagcttta atgcgcggcg
     1381 caagctagcc ggagccatac ttttcacaat gttggctacc agaaattttt cggagctggc
     1441 cgagaagctg gccggcctgg acatcaacgg tggagctagc ggcagccggg cgaccaccct
     1501 ggaacgtatt gaaaagagct ttgtgattac cgatccgcgt ctgccggata acccgattat
     1561 ttttgtgagc gatagctttc tgcagctgac cgaatatagc cgtgaagaaa ttctgggccg
     1621 taactgccgt tttctgcagg gcccggaaac cgatcgtgcg accgtgcgta aaattcgtga
     1681 tgcgattgat aaccagaccg aagtgaccgt gcagctgatt aactatacca aaagcggcaa
     1741 aaaattttgg aacctgtttc atctgcagcc gatgcgtgat cagaaaggcg atgtgcagta
     1801 ttttattggc gtgcagctgg atggtaccga acgtgtgcgt gatgcggcgg aacgtgaagc
     1861 tgtgatgctg gtcaagaaaa ccgcggagga aatcgacgaa gccgctaagg aaaatctcta
     1921 cttccagatg ggcggaggat ccgactacaa ggatgacgac gataagatgt ctagattaga
     1981 taaaagtaaa gtgattaaca gcgcattaga gctgcttaat gaggtcggaa tcgaaggttt
     2041 aacaacccgt aaactcgccc agaagctagg tgtagagcag cctacattgt attggcatgt
     2101 aaaaaataag cgggctttgc tcgacgcctt agccattgag atgttagata ggcaccatac
     2161 tcacttttgc cctttagaag gggaaagctg gcaagatttt ttacgtaata acgctaaaag
     2221 ttttagatgt gctttactaa gtcatcgcga tggagcaaaa gtacatttag gtacacggcc
     2281 tacagaaaaa cagtatgaaa ctctcgaaaa tcaattagcc tttttatgcc aacaaggttt
     2341 ttcactagag aatgcattat atgcactcag cgctgtgggg cattttactt taggttgcgt
     2401 attggaagat caagagcatc aagtcgctaa agaagaaagg gaaacaccta ctactgatag
     2461 tatgccgcca ttattacgac aagctatcga attatttgat caccaaggtg cagagccagc
     2521 cttcttattc ggccttgaat tgatcatatg cggattagaa aaacaactta aatgtgaaag
     2581 tgggtccgcg tacagccgcg cgcgtacgaa aaacaattac gggtctacca tcgagggcct
     2641 gctcgatctc ccggacgacg acgcccccga agaggcgggg ctggcggctc cgcgcctgtc
     2701 ctttctcccc gcgggacaca cgcgcagact gtcgacggcc cccccgaccg atgtcagcct
     2761 gggggacgag ctccacttag acggcgagga cgtggcgatg gcgcatgccg acgcgctaga
     2821 cgatttcgat ctggacatgt tgggggacgg ggattccccg ggtccgggat ttacccccca
     2881 cgactccgcc ccctacggcg ctctggatat ggccgacttc gagtttgagc agatgtttac
     2941 cgatgccctt ggaattgacg agtacggtgg gtaataggaa ttcctcgagc tagagctcgc
     3001 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     3061 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     3121 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     3181 aagggggagg attgggaaga caatagcagg catgctgggg aggtaaccac gtgcggaccg
     3241 agcggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc
     3301 tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag
     3361 tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc
     3421 atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg
     3481 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     3541 cttagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     3601 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     3661 cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac
     3721 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     3781 tggaacaaca ctcaactcta tctcgggcta ttcttttgat ttataaggga ttttgccgat
     3841 ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     3901 aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg atgccgcata
     3961 gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     4021 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     4081 ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata
     4141 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     4201 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     4261 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     4321 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     4381 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     4441 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     4501 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     4561 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     4621 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     4681 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     4741 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     4801 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     4861 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     4921 aatagactgg atggaggtgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     4981 tggctggttt attgctgata aatctggagc cggtgagcgt ggaagccgcg gtatcattgc
     5041 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     5101 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     5161 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     5221 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     5281 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     5341 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     5401 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     5461 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     5521 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     5581 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     5641 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     5701 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     5761 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     5821 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     5881 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     5941 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgtcct gcaggcagct
     6001 gcgcgctcgc tcgctcactg aggccgcccg ggcgtcgggc gacctttggt cgcccggcct
     6061 cagtgagcga gcgagcgcgc agagagggag tggccaactc catcactagg ggttcctgcg
     6121 gccgcacgcg ttgtacagtg tctagactgc agagggccct gcgtatgagt gcaagtgggt
     6181 tttaggacca ggatgaggcg gggtgggggt gcctacctga cgaccgaccc cgacccactg
     6241 gacaagcacc caacccccat tccccaaatt gcgcatcccc tatcagagag ggggagggga
     6301 aacaggatgc ggcgaggcgc gtgcgcactg ccagcttcag caccgcggac agtgccttcg
     6361 cccccgcctg gcggc
//
