LOCUS       sequence-320936-        6228 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FLiCRE protease for neuronal expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6228)
  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 6228)
  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: mtagBFP2-CaM-uTEVp
FEATURES             Location/Qualifiers
     source          1..6228
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        181..628
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      642..651
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             648..1358
                     /codon_start=1
                     /product="enhanced monomeric blue fluorescent protein 
                     (Subach et al., 2011)"
                     /label=mTagBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRND
                     MALKLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
     CDS             1395..1838
                     /codon_start=1
                     /product="calcium-binding protein"
                     /label=calmodulin
                     /translation="MDQLTEEQIAEFKEAFSLLDKDGDGTITTKELGTGMRSLGQNPTE
                     AELQDMINEVDADGDGTIDFPEFLTMMARKMKYTDSEEEIREAFRVFDKDGNGYISAAE
                     LRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     CDS             1839..1880
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    2619..3207
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2672..2692)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3090..3101)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3249..3266)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3255..3462
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3491..3631
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3491..3620
                     /label=AAV2 ITR
     rep_origin      3706..4161
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3793..3812)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4003..4024
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4178..4197)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4297..4319
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4357..4375)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4443..4547
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4548..4616,4617..5408)
                     /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     complement(4766..4785)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5579..6167
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6068..6087
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgttgtaca gtgtctagac tgcagagggc cctgcgtatg
      181 agtgcaagtg ggttttagga ccaggatgag gcggggtggg ggtgcctacc tgacgaccga
      241 ccccgaccca ctggacaagc acccaacccc cattccccaa attgcgcatc ccctatcaga
      301 gagggggagg ggaaacagga tgcggcgagg cgcgtgcgca ctgccagctt cagcaccgcg
      361 gacagtgcct tcgcccccgc ctggcggcgc gcgccaccgc cgcctcagca ctgaaggcgc
      421 gctgacgtca ctcgccggtc ccccgcaaac tccccttccc ggccaccttg gtcgcgtccg
      481 cgccgccgcc ggcccagccg gaccgcacca cgcgaggcgc gagatagggg ggcacgggcg
      541 cgaccatctg cgctgcggcg ccggcgactc agcgctgcct cagtctgcgg tgggcagcgg
      601 aggagtcgtg tcgtgcctga gagcgcagtc gagaaggatc cgccaccatg gtgtctaagg
      661 gcgaagagct gattaaggag aacatgcaca tgaagctgta catggagggc accgtggaca
      721 accatcactt caagtgcaca tccgagggcg aaggcaagcc ctacgagggc acccagacca
      781 tgagaatcaa ggtggtcgag ggcggccctc tccccttcgc cttcgacatc ctggctacta
      841 gcttcctcta cggcagcaag accttcatca accacaccca gggcatcccc gacttcttca
      901 agcagtcctt ccctgagggc ttcacatggg agagagtcac cacatacgaa gacgggggcg
      961 tgctgaccgc tacccaggac accagcctcc aggacggctg cctcatctac aacgtcaaga
     1021 tcagaggggt gaacttcaca tccaacggcc ctgtgatgca gaagaaaaca ctcggctggg
     1081 aggccttcac cgagacgctg taccccgctg acggcggcct ggaaggcaga aacgacatgg
     1141 ccctgaagct cgtgggcggg agccatctga tcgcaaacgc caagaccaca tatagatcca
     1201 agaaacccgc taagaacctc aagatgcctg gcgtctacta tgtggactac agactggaaa
     1261 gaatcaagga ggccaacaac gagacctacg tcgagcagca cgaggtggca gtggccagat
     1321 actgcgacct ccctagcaaa ctggggcaca agcttaatga ccttggatct ggcacaggcg
     1381 gatccggttc aggcatggac caactgactg aagagcagat cgcagaattt aaagaggctt
     1441 tctccctact ggacaaggac ggggatggga caataacaac caaggagctg gggacgggga
     1501 tgcggtctct ggggcagaac cccacagaag cagagctgca ggacatgatc aatgaagtag
     1561 atgccgacgg tgacggcaca atcgacttcc ctgagttcct gacaatgatg gcaagaaaaa
     1621 tgaaatacac agacagtgaa gaagaaatta gagaagcgtt ccgtgtgttt gataaggatg
     1681 gcaatggcta catcagtgca gcagagcttc gccacgtgat gacaaacctt ggagagaagt
     1741 taacagatga agaggttgat gaaatgatca gggaagcaga catcgatggg gatggtcagg
     1801 taaactacga agagtttgta caaatgatga cagcgaaggg caagccgatc cctaaccctc
     1861 tgctgggcct ggacagcact ggtggaagtg gatcaggcag cggtggatct tacggaagcc
     1921 atgtggatta cgccggagaa tccttgttta agggaccacg tgattacaac ccgatatcga
     1981 gcaccatttg tcatttgacg aatgaatctg atgggcacac aacatcgttg tatggtattg
     2041 gatttggtcc cttcatcatt acaaacaagc acttgtttag aagaaataat ggaacactgt
     2101 tggtccaatc actacatggt gtattcaagg tcaagaacac cacgactttg caacaacacc
     2161 tcattgatgg gagggacatg ataattattc gcatgcctaa ggatttccca ccatttcctc
     2221 aaaagctgaa atttagagag ccacaaaggg aagagcgcat atgtcttgtg acaaccaact
     2281 tccaaactaa gagcatgtct agcatggtgt cagacactag ttgcacattc ccttcatctg
     2341 atggcatatt ctggaagcat tggattcaaa ccaaggatgg gcagtgtggc aatccattag
     2401 tatcaactag agatgggttc attgttggta tacactcagc atcgaatttc accaacacaa
     2461 acaattattt cacaagcgtg ccgaaaaact tcatggaatt gttgacaaat caggaggcgc
     2521 agcagtgggt tagtggttgg cgattaaatg ctgactcagt attgtggggg ggccataaag
     2581 ttttcatggt gtaagaattc gatatcaagc ttatcgataa tcaacctctg gattacaaaa
     2641 tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg
     2701 ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct
     2761 tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg
     2821 gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct
     2881 gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg
     2941 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg
     3001 tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctatgttgcc acctggattc
     3061 tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc
     3121 gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc
     3181 ggatctccct ttgggccgcc tccccgcatc gataccgagc gctgctcgag ctagagctcg
     3241 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     3301 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     3361 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     3421 caagggggag gattgggaag acaatagcag gcatgctggg gaggtaacca cgtgcggacc
     3481 gagcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg
     3541 ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca
     3601 gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg
     3661 catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg
     3721 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     3781 ccttagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     3841 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     3901 tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga
     3961 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     4021 ctggaacaac actcaactct atctcgggct attcttttga tttataaggg attttgccga
     4081 tttcggtcta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     4141 aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat
     4201 agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     4261 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     4321 tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat
     4381 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
     4441 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     4501 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     4561 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     4621 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     4681 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     4741 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     4801 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     4861 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     4921 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     4981 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     5041 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     5101 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     5161 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     5221 ctggctggtt tattgctgat aaatctggag ccggtgagcg tggaagccgc ggtatcattg
     5281 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     5341 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     5401 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     5461 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     5521 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     5581 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     5641 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     5701 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     5761 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     5821 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     5881 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     5941 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     6001 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     6061 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     6121 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     6181 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgt
//
