LOCUS       sequence-315101-        6327 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Reporter construct.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6327)
  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 6327)
  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: TRE-mCherry-p2A-Chrimson-TS
FEATURES             Location/Qualifiers
     source          1..6327
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..39
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     36..60
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      88..97
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             94..801
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(239..257)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     507..526
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             811..867
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             868..1917
                     /codon_start=1
                     /product="red light-drivable channelrhodopsin from 
                     Chlamydomonas noctigama (Klapoetke et al., 2014)"
                     /label=ChrimsonR
                     /note="K176R mutant with accelerated closing kinetics"
                     /translation="MAELISSATRSLFAAGGINPWPNPYHHEDMGCGGMTPTGECFSTE
                     WWCDPSYGLSDAGYGYCFVEATGGYLVVGVEKKQAWLHSRGTPGEKIGAQVCQWIAFSI
                     AIALLTFYGFSAWKATCGWEEVYVCCVEVLFVTLEIFKEFSSPATVYLSTGNHAYCLRY
                     FEWLLSCPVILIRLSNLSGLKNDYSKRTMGLIVSCVGMIVFGMAAGLATDWLKWLLYIV
                     SCIYGGYMYFQAAKCYVEANHSVPKGHCRMVVKLMAYAYFASWGSYPILWAVGPEGLLK
                     LSPYANSIGHSICDIIAKEFWTFLAHHLRIKIHEHILIHGDIRKTTKMEIGGEEVEVEE
                     FVEEEDEDTV"
     CDS             2026..2052
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    2071..2659
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2124..2144)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2542..2553)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2691..3167
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2890..2909)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3207..3347
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3207..3336
                     /label=AAV2 ITR
     rep_origin      3422..3877
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3509..3528)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3719..3740
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3894..3913)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4013..4035
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4073..4091)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4159..4263
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4264..4332,4333..5124)
                     /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(4482..4501)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5295..5883
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5784..5803
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   5945..6074
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     protein_bind    6120..6138
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6155..6173
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6191..6209
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6227..6245
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6262..6280
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6298..6316
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
ORIGIN
        1 ggtaggcgtg tacggtggga ggcctatata agcagagctc gtttagtgaa ccgtcagatc
       61 gcaaagggcg aattcgatcc accggtcgcc accatggtga gcaagggcga ggaggataac
      121 atggccatca tcaaggagtt catgcgcttc aaggtgcaca tggagggctc cgtgaacggc
      181 cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc
      241 aagctgaagg tgaccaaggg tggccccctg cccttcgcct gggacatcct gtcccctcag
      301 ttcatgtacg gctccaaggc ctacgtgaag caccccgccg acatccccga ctacttgaag
      361 ctgtccttcc ccgagggatt caagtgggag cgcgtgatga acttcgagga cggcggcgtg
      421 gtgaccgtga cccaggactc ctccctgcag gacggcgagt tcatctacaa ggtgaagctg
      481 cgcggcacca acttcccctc cgacggcccc gtaatgcaga agaagaccat gggctgggag
      541 gcctcctccg agcggatgta ccccgaggac ggcgccctga agggcgagat caagcagagg
      601 ctgaagctga aggacggcgg ccactacgac gctgaggtca agaccaccta caaggccaag
      661 aagcccgtgc agctgcccgg cgcctacaac gtcaacatca agttggacat cacctcccac
      721 aacgaggact acaccatcgt ggaacagtac gaacgcgccg agggccgcca ctccaccggc
      781 ggcatggacg agctgtacaa gggctccgga gccacgaact tctctctgtt aaagcaagca
      841 ggagacgtgg aagaaaaccc cggtcccatg gctgagctga tcagcagcgc caccagatct
      901 ctgtttgccg ccggaggcat caacccttgg cctaacccct accaccacga ggacatgggc
      961 tgtggaggaa tgacacctac aggcgagtgc ttcagcaccg agtggtggtg tgacccttct
     1021 tacggactga gcgacgccgg atacggatat tgcttcgtgg aggccacagg cggctacctg
     1081 gtcgtgggag tggagaagaa gcaggcttgg ctgcacagca gaggcacacc aggagaaaag
     1141 atcggcgccc aggtctgcca gtggattgct ttcagcatcg ccatcgccct gctgacattc
     1201 tacggcttca gcgcctggaa ggccacttgc ggttgggagg aggtctacgt ctgttgcgtc
     1261 gaggtgctgt tcgtgaccct ggagatcttc aaggagttca gcagccccgc cacagtgtac
     1321 ctgtctaccg gcaaccacgc ctattgcctg cgctacttcg agtggctgct gtcttgcccc
     1381 gtgatcctga tcagactgag caacctgagc ggcctgaaga acgactacag caagcggacc
     1441 atgggcctga tcgtgtcttg cgtgggaatg atcgtgttcg gcatggccgc aggactggct
     1501 accgattggc tcaagtggct gctgtatatc gtgtcttgca tctacggcgg ctacatgtac
     1561 ttccaggccg ccaagtgcta cgtggaagcc aaccacagcg tgcctaaagg ccattgccgc
     1621 atggtcgtga agctgatggc ctacgcttac ttcgcctctt ggggcagcta cccaatcctc
     1681 tgggcagtgg gaccagaagg actgctgaag ctgagccctt acgccaacag catcggccac
     1741 agcatctgcg acatcatcgc caaggagttt tggaccttcc tggcccacca cctgaggatc
     1801 aagatccacg agcacatcct gatccacggc gacatccgga agaccaccaa gatggagatc
     1861 ggaggcgagg aggtggaagt ggaagagttc gtggaggagg aggacgagga cacagtggcg
     1921 gcaccggtag tagcagtgag cgatgcggcc gccaagagca ggatcaccag cgagggcgag
     1981 tacatccccc tggaccagat cgacatcaac gtgggcgcgc ccgggtaccc atacgatgtg
     2041 ccagattacg cctaatagaa gcttatcgat aatcaacctc tggattacaa aatttgtgaa
     2101 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     2161 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     2221 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
     2281 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
     2341 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
     2401 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
     2461 gggaaatcat cgtcctttcc ttggctgctc gcctatgttg ccacctggat tctgcgcggg
     2521 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
     2581 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
     2641 ctttgggccg cctccccgca tcgataccga gcgctgctcg agagatctac gggtggcatc
     2701 cctgtgaccc ctccccagtg cctctcctgg ccctggaagt tgccactcca gtgcccacca
     2761 gccttgtcct aataaaatta agttgcatca ttttgtctga ctaggtgtcc ttctataata
     2821 ttatggggtg gaggggggtg gtatggagca aggggcaagt tgggaagaca acctgtaggg
     2881 cctgcggggt ctattgggaa ccaagctgga gtgcagtggc acaatcttgg ctcactgcaa
     2941 tctccgcctc ctgggttcaa gcgattctcc tgcctcagcc tcccgagttg ttgggattcc
     3001 aggcatgcat gaccaggctc agctaatttt tgtttttttg gtagagacgg ggtttcacca
     3061 tattggccag gctggtctcc aactcctaat ctcaggtgat ctacccacct tggcctccca
     3121 aattgctggg attacaggcg tgaaccactg ctcccttccc tgtccttctg attttgtagg
     3181 taaccacgtg cggaccgagc ggccgcagga acccctagtg atggagttgg ccactccctc
     3241 tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt
     3301 tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg
     3361 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag
     3421 tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     3481 gctacacttg ccagcgcctt agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     3541 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     3601 agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg
     3661 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     3721 ggactcttgt tccaaactgg aacaacactc aactctatct cgggctattc ttttgattta
     3781 taagggattt tgccgatttc ggtctattgg ttaaaaaatg agctgattta acaaaaattt
     3841 aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc
     3901 tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc
     3961 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     4021 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg
     4081 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     4141 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     4201 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     4261 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     4321 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     4381 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     4441 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     4501 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     4561 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     4621 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     4681 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     4741 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     4801 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     4861 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     4921 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     4981 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     5041 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     5101 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     5161 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     5221 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     5281 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     5341 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     5401 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     5461 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     5521 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     5581 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     5641 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     5701 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     5761 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     5821 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     5881 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     5941 atgtcctgca ggcagctgcg cgctcgctcg ctcactgagg ccgcccgggc gtcgggcgac
     6001 ctttggtcgc ccggcctcag tgagcgagcg agcgcgcaga gagggagtgg ccaactccat
     6061 cactaggggt tcctgcggcc gcacgcgtcc ttcaggtccg aggttctaga cgagtttact
     6121 ccctatcagt gatagagaac gatgtcgagt ttactcccta tcagtgatag agaacgtatg
     6181 tcgagtttac tccctatcag tgatagagaa cgtatgtcga gtttactccc tatcagtgat
     6241 agagaacgta tgtcgagttt atccctatca gtgatagaga acgtatgtcg agtttactcc
     6301 ctatcagtga tagagaacgt atgtcga
//
