LOCUS       sequence-315100-        6165 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 6165)
  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 6165)
  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-bReaches-TS
FEATURES             Location/Qualifiers
     source          1..6165
                     /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             1864..1890
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    1909..2497
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1962..1982)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2380..2391)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2529..3005
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2728..2747)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3045..3185
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3045..3174
                     /label=AAV2 ITR
     rep_origin      3260..3715
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3347..3366)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3557..3578
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3732..3751)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3851..3873
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3911..3929)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3997..4101
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4102..4170,4171..4962)
                     /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(4320..4339)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5133..5721
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5622..5641
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   5783..5912
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     protein_bind    5958..5976
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5993..6011
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6029..6047
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6065..6083
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6100..6118
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6136..6154
                     /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 gactatggcg gcgctttgtc tgccgtcgga
      901 ctgttccaga ccagctacac cctggagaac aacggcagcg tgatctgcat ccccaacaac
      961 ggccagtgct tctgcctggc ctggctgaag agtaacggca ccaacgccga gaagctggcc
     1021 gccaacatcc tgcagtgggt ggtgtttgcg ctgagcgtgg cgtgcctggg ctggtatgcg
     1081 tatcaggcgt ggcgcgcgac ctgcggctgg gaagaagtgt atgtggcgct gattgaaatg
     1141 atgaaaagca ttattgaagc gtttcatgaa tttgatagcc cggcgaccct gtggctgagc
     1201 agcggcaacg gcgtggtgtg gatgcgctat ggctcatggc tgctgacctg ccccgtgatt
     1261 ctgattcatc tgagcaacct gaccggcctg aaagatgatt atagcaaacg caccatgggc
     1321 ctgctggtga gcgacgtggg ctgcattgtg tggggcgcga ccagcgcgat gtgcaccggc
     1381 tggaccaaaa ttctgttttt tctgattagc ctgagctatg gcatgtatac ctattttcat
     1441 gcggccaaag tgtatattga agcgtttcat accgtgccga aaggcctgtg cagacagctg
     1501 gtgagagcca tggcctggct gttcttcgtg agctggggca tgttccccgt gctgttcctg
     1561 ctgggccccg agggcttcgg ccatattagc ccgtatggca gcgcgattgg ccatagcatt
     1621 ctggatctga ttgcgaagaa catgtggggc gtgctgggca actatctgcg cgtgaaaatt
     1681 catgaacata ttctgctgta tggcgatatt cgcaaaaaac agaaaattac cattgcgggc
     1741 caggaaatgg aagtggaaac cctggtggcg gaagaagaag atgcggccgc caagagcagg
     1801 atcaccagcg agggcgagta catccccctg gaccagatcg acatcaacgt gggcgcgccc
     1861 gggtacccat acgatgtgcc agattacgcc taatagaagc ttatcgataa tcaacctctg
     1921 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     1981 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     2041 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
     2101 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
     2161 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
     2221 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     2281 aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctatgttgcc
     2341 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
     2401 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
     2461 cagacgagtc ggatctccct ttgggccgcc tccccgcatc gataccgagc gctgctcgag
     2521 agatctacgg gtggcatccc tgtgacccct ccccagtgcc tctcctggcc ctggaagttg
     2581 ccactccagt gcccaccagc cttgtcctaa taaaattaag ttgcatcatt ttgtctgact
     2641 aggtgtcctt ctataatatt atggggtgga ggggggtggt atggagcaag gggcaagttg
     2701 ggaagacaac ctgtagggcc tgcggggtct attgggaacc aagctggagt gcagtggcac
     2761 aatcttggct cactgcaatc tccgcctcct gggttcaagc gattctcctg cctcagcctc
     2821 ccgagttgtt gggattccag gcatgcatga ccaggctcag ctaatttttg tttttttggt
     2881 agagacgggg tttcaccata ttggccaggc tggtctccaa ctcctaatct caggtgatct
     2941 acccaccttg gcctcccaaa ttgctgggat tacaggcgtg aaccactgct cccttccctg
     3001 tccttctgat tttgtaggta accacgtgcg gaccgagcgg ccgcaggaac ccctagtgat
     3061 ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt
     3121 cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct
     3181 gcaggggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     3241 acgtcaaagc aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     3301 gttacgcgca gcgtgaccgc tacacttgcc agcgccttag cgcccgctcc tttcgctttc
     3361 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     3421 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt
     3481 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     3541 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ctctatctcg
     3601 ggctattctt ttgatttata agggattttg ccgatttcgg tctattggtt aaaaaatgag
     3661 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aattttatgg
     3721 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca
     3781 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     3841 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     3901 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     3961 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     4021 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     4081 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     4141 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     4201 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     4261 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     4321 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     4381 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     4441 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     4501 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     4561 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     4621 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     4681 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     4741 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     4801 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     4861 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     4921 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     4981 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     5041 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     5101 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     5161 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     5221 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     5281 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     5341 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     5401 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     5461 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     5521 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     5581 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     5641 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     5701 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     5761 tgctggcctt ttgctcacat gtcctgcagg cagctgcgcg ctcgctcgct cactgaggcc
     5821 gcccgggcgt cgggcgacct ttggtcgccc ggcctcagtg agcgagcgag cgcgcagaga
     5881 gggagtggcc aactccatca ctaggggttc ctgcggccgc acgcgtcctt caggtccgag
     5941 gttctagacg agtttactcc ctatcagtga tagagaacga tgtcgagttt actccctatc
     6001 agtgatagag aacgtatgtc gagtttactc cctatcagtg atagagaacg tatgtcgagt
     6061 ttactcccta tcagtgatag agaacgtatg tcgagtttat ccctatcagt gatagagaac
     6121 gtatgtcgag tttactccct atcagtgata gagaacgtat gtcga
//
