LOCUS       sequence-327803-        6364 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FLiCRE TF with fast kinetics for neuronal expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6364)
  AUTHORS   Kim CK, Sanchez MI, Hoerbelt P, Fenno LE, Malenka RC, Deisseroth K, 
            Ting AY
  TITLE     A Molecular Calcium Integrator Reveals a Striatal Cell Type Driving 
            Aversion.
  JOURNAL   Cell. 2020 Dec 23;183(7):2003-2019.e16. doi: 
            10.1016/j.cell.2020.11.015. Epub 2020 Dec 11.
  PUBMED    33308478
REFERENCE   2  (bases 1 to 6364)
  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"; doi: 
            "10.1016/j.cell.2020.11.015"; journalName: "Cell"; date: 
            "2020-12-23- 23"; volume: "183"; issue: "7"; pages: "2003-2019.e16"
COMMENT     Sequence Label: Nrxn3b-Nav1.6-MKII-f-hLOV1-TEVcs(ENLYFQ/M)-tTA...
FEATURES             Location/Qualifiers
     source          1..6364
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(1..69,70..861)
                     /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(219..238)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1032..1620
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1521..1540
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   1682..1811
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        1862..2309
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             2492..2518
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2744..3253
                     /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             3968..3988
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQM"
     CDS             4001..4024
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             join(4025..4645,4646..5032)
                     /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(4098..4117)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     primer_bind     complement(5066..5083)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5072..5279
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5308..5448
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5308..5437
                     /label=AAV2 ITR
     rep_origin      5523..5978
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5610..5629)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5820..5841
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5995..6014)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6114..6136
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6174..6192)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6260..6364
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
       61 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
      121 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
      181 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
      241 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
      301 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
      361 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
      421 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
      481 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
      541 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
      601 tcccggcaac aattaataga ctggatggag gtggataaag ttgcaggacc acttctgcgc
      661 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtggaagc
      721 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
      781 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
      841 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
      901 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
      961 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     1021 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     1081 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     1141 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     1201 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     1261 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     1321 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     1381 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     1441 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     1501 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     1561 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     1621 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     1681 tcctgcaggc agctgcgcgc tcgctcgctc actgaggccg cccgggcgtc gggcgacctt
     1741 tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca actccatcac
     1801 taggggttcc tgcggccgca cgcgttgtac agtgtctaga ctgcagaggg ccctgcgtat
     1861 gagtgcaagt gggttttagg accaggatga ggcggggtgg gggtgcctac ctgacgaccg
     1921 accccgaccc actggacaag cacccaaccc ccattcccca aattgcgcat cccctatcag
     1981 agagggggag gggaaacagg atgcggcgag gcgcgtgcgc actgccagct tcagcaccgc
     2041 ggacagtgcc ttcgcccccg cctggcggcg cgcgccaccg ccgcctcagc actgaaggcg
     2101 cgctgacgtc actcgccggt cccccgcaaa ctccccttcc cggccacctt ggtcgcgtcc
     2161 gcgccgccgc cggcccagcc ggaccgcacc acgcgaggcg cgagataggg gggcacgggc
     2221 gcgaccatct gcgctgcggc gccggcgact cagcgctgcc tcagtctgcg gtgggcagcg
     2281 gaggagtcgt gtcgtgcctg agagcgcagt cgagaagaag cttgccacca tgcacctgag
     2341 aatccacgcg agacggagcc ctcctcgccg gccggcctgg acgcttggga tctggttcct
     2401 gttctgggga tgtatcgtca gctctgtatg gagtcaattg tcttctaatg tagcttcctc
     2461 ctcctccacc tcttcctcgc cggggtctca ctacccatac gatgtgccag attacgccaa
     2521 ccccacggag ccgggaatca gacgggttcc gggggcctca gaggtgatcc gggagtcgag
     2581 cagcacaaca gggatggtcg tcggcattgt ggctgctgcc gccctctgca tcttgatcct
     2641 cctgtacgcc atgtacaagt acaggaacag ggacgagggc tcgggctcga cctcgggctc
     2701 gggcagcggt ggctctggag gttctggtgg cagctctgga ggtatgaatg gagctatagg
     2761 aggtgacctt ttgctcaatt ttcctgacat gtcggtccta gagcgccaaa gggctcacct
     2821 caagtacctc aatcccacct ttgattctcc tctcgccggc ttctttgccg attcttcaat
     2881 gattaccggc ggcgagatgg acagctatct ttcgactgcc ggtttgaatc ttccgatgat
     2941 gtacggtgag acgacggtgg aaggtgattc aagactctca atttcgccgg aaacgacgct
     3001 tgggactgga aatttcaagg cagcgaagtt tgatacagag actaaggatt gtaatgaggc
     3061 ggcgaagaag atgacgatga acagagatga cctagtagaa gaaggagaag aagagaagtc
     3121 gaaaataaca gagcaaaaca atgggagcac aaaaagcatc aagaagatga aacacaaagc
     3181 caagaaagaa gagaacaatt tctctaatga ttcatctaaa gtgacgaagg aattggagaa
     3241 aacggattat attcatagcg gtagtggcac cgtgcgcgtg ccgattgcgg tgggcgaaag
     3301 cgattttgaa aacctgaaca ccgaagatgt gagcagcgaa agcgatccgg gtggctctgg
     3361 tagcttcaac gctcgcagga agctggctgg tgctatcctg ttcaccatgc tcgccacaag
     3421 aaacttcagc ggcagcttta atgcgcggcg caagctggcc ggagccatac ttttcacaat
     3481 gttggctacc agaaattttt cggagctggc cgagaagctg gccggcctgg acatcaacgg
     3541 tggagctagc gagtttcggg caaccacact ggaacggatc gagaaaagct tcgtgattac
     3601 tgatccgaga ctgcctgaca acccagtgat ttttgtgagc gattccttcc tgcagctgac
     3661 agaatattct cgggaagaga tcctggggcg caattgccgt tttctgcagg gacccgagac
     3721 agaccgtgcc actgttcgga aaatcagaga tgctattgac aaccagactg aagtgaccgt
     3781 tcagctgatc aattatacca agagcggcaa gaagttctgg aacgtgttcc acctgcagcc
     3841 gatgcgcgat tataagggcg acgtccagta cttcattggc gtgcagctgg atggcaccga
     3901 acgtcttcat ggcgccgctg agcgtgaggc ggtctgcctg gtcaaaaaga cagcctttca
     3961 gattgctgag aacctgtact tccagatggg cggaggatcc gactacaagg atgacgacga
     4021 taagatgtct agattagata aaagtaaagt gattaacagc gcattagagc tgcttaatga
     4081 ggtcggaatc gaaggtttaa caacccgtaa actcgcccag aagctaggtg tagagcagcc
     4141 tacattgtat tggcatgtaa aaaataagcg ggctttgctc gacgccttag ccattgagat
     4201 gttagatagg caccatactc acttttgccc tttagaaggg gaaagctggc aagatttttt
     4261 acgtaataac gctaaaagtt ttagatgtgc tttactaagt catcgcgatg gagcaaaagt
     4321 acatttaggt acacggccta cagaaaaaca gtatgaaact ctcgaaaatc aattagcctt
     4381 tttatgccaa caaggttttt cactagagaa tgcattatat gcactcagcg ctgtggggca
     4441 ttttacttta ggttgcgtat tggaagatca agagcatcaa gtcgctaaag aagaaaggga
     4501 aacacctact actgatagta tgccgccatt attacgacaa gctatcgaat tatttgatca
     4561 ccaaggtgca gagccagcct tcttattcgg ccttgaattg atcatatgcg gattagaaaa
     4621 acaacttaaa tgtgaaagtg ggtccgcgta cagccgcgcg cgtacgaaaa acaattacgg
     4681 gtctaccatc gagggcctgc tcgatctccc ggacgacgac gcccccgaag aggcggggct
     4741 ggcggctccg cgcctgtcct ttctccccgc gggacacacg cgcagactgt cgacggcccc
     4801 cccgaccgat gtcagcctgg gggacgagct ccacttagac ggcgaggacg tggcgatggc
     4861 gcatgccgac gcgctagacg atttcgatct ggacatgttg ggggacgggg attccccggg
     4921 tccgggattt accccccacg actccgcccc ctacggcgct ctggatatgg ccgacttcga
     4981 gtttgagcag atgtttaccg atgcccttgg aattgacgag tacggtgggt aataggaatt
     5041 cctcgagcta gagctcgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     5101 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     5161 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     5221 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggag
     5281 gtaaccacgt gcggaccgag cggccgcagg aacccctagt gatggagttg gccactccct
     5341 ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct
     5401 ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc
     5461 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata
     5521 gtacgcgccc trtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     5581 cgctacactt gccagcgcct tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     5641 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     5701 tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg
     5761 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     5821 tggactcttg ttccaaactg gaacaacact caactctatc tcgggctatt cttttgattt
     5881 ataagggatt ttgccgattt cggtctattg gttaaaaaat gagctgattt aacaaaaatt
     5941 taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat
     6001 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc
     6061 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     6121 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt
     6181 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     6241 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     6301 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     6361 gagt
//
