LOCUS       sequence-318318-        6123 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Reporter construct.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6123)
  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 6123)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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-eNpHR3.0-TS
FEATURES             Location/Qualifiers
     source          1..6123
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     protein_bind    176..194
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    211..229
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    247..265
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    283..301
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    318..336
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    354..372
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        384..422
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     419..443
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      471..480
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             477..1184
                     /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(622..640)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     890..909
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1194..1250
                     /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             2205..2231
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    2250..2838
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2303..2323)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2721..2732)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2870..3346
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3069..3088)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3386..3526
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3386..3515
                     /label=AAV2 ITR
     rep_origin      3601..4056
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3688..3707)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3898..3919
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4073..4092)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4192..4214
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4252..4270)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4338..4442
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4443..4511,4512..5303)
                     /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(4661..4680)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5474..6062
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5963..5982
                     /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 gcgtccttca ggtccgaggt tctagacgag tttactccct
      181 atcagtgata gagaacgatg tcgagtttac tccctatcag tgatagagaa cgtatgtcga
      241 gtttactccc tatcagtgat agagaacgta tgtcgagttt actccctatc agtgatagag
      301 aacgtatgtc gagtttatcc ctatcagtga tagagaacgt atgtcgagtt tactccctat
      361 cagtgataga gaacgtatgt cgaggtaggc gtgtacggtg ggaggcctat ataagcagag
      421 ctcgtttagt gaaccgtcag atcgcaaagg gcgaattcga tccaccggtc gccaccatgg
      481 tgagcaaggg cgaggaggat aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc
      541 acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc
      601 cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttcg
      661 cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg aagcaccccg
      721 ccgacatccc cgactacttg aagctgtcct tccccgaggg attcaagtgg gagcgcgtga
      781 tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg caggacggcg
      841 agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc
      901 agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag gacggcgccc
      961 tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac gacgctgagg
     1021 tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca
     1081 tcaagttgga catcacctcc cacaacgagg actacaccat cgtggaacag tacgaacgcg
     1141 ccgagggccg ccactccacc ggcggcatgg acgagctgta caagggctcc ggagccacga
     1201 acttctctct gttaaagcaa gcaggagacg tggaagaaaa ccccggtccc atgacagaga
     1261 ccctgcctcc cgtgaccgag agtgccgtgg cccttcaagc cgaggttacc caaagggagt
     1321 tgttcgagtt cgtgctgaac gaccctttgc ttgcaagcag tctctatatc aacatcgcac
     1381 ttgcaggact gagtatactg ctgttcgttt ttatgacccg aggactcgat gatccacggg
     1441 caaaacttat tgctgtgtca accatccttg tgcctgtcgt cagcattgcc tcctacactg
     1501 gattggcgag cggcctgaca atttccgttc ttgaaatgcc agcgggccat tttgcagaag
     1561 gcagctcagt gatgctggga ggagaagagg tagatggtgt agtcaccatg tggggacggt
     1621 atctcacctg ggcactttcc acgcccatga ttctcctcgc tctgggtctc ctggccggaa
     1681 gcaatgctac aaagctcttc acagctatca ctttcgatat cgctatgtgc gtgactggcc
     1741 ttgccgcggc cctgactacc tcctcccacc tcatgagatg gttctggtac gctatcagtt
     1801 gtgcatgctt tctggtggtc ttgtatatcc tgctggtgga gtgggcacag gacgccaaag
     1861 ccgcgggaac cgctgacatg ttcaataccc tgaagctgtt gacagtagtg atgtggctgg
     1921 ggtatccaat tgtgtgggct cttggagtcg agggtatcgc ggtgttgccc gttggggtga
     1981 cgagctgggg atattctttc ctggatatcg tggcaaagta cattttcgca ttcttgctcc
     2041 tgaactatct gacgtcaaac gaatctgtcg tgtccggcag cattttggat gttccatctg
     2101 cttctgggac cccggctgat gatgcggccg ccaagagcag gatcaccagc gagggcgagt
     2161 acatccccct ggaccagatc gacatcaacg tgggcgcgcc cgggtaccca tacgatgtgc
     2221 cagattacgc ctaatagaag cttatcgata atcaacctct ggattacaaa atttgtgaaa
     2281 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     2341 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     2401 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     2461 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     2521 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     2581 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     2641 ggaaatcatc gtcctttcct tggctgctcg cctatgttgc cacctggatt ctgcgcggga
     2701 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     2761 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
     2821 tttgggccgc ctccccgcat cgataccgag cgctgctcga gagatctacg ggtggcatcc
     2881 ctgtgacccc tccccagtgc ctctcctggc cctggaagtt gccactccag tgcccaccag
     2941 ccttgtccta ataaaattaa gttgcatcat tttgtctgac taggtgtcct tctataatat
     3001 tatggggtgg aggggggtgg tatggagcaa ggggcaagtt gggaagacaa cctgtagggc
     3061 ctgcggggtc tattgggaac caagctggag tgcagtggca caatcttggc tcactgcaat
     3121 ctccgcctcc tgggttcaag cgattctcct gcctcagcct cccgagttgt tgggattcca
     3181 ggcatgcatg accaggctca gctaattttt gtttttttgg tagagacggg gtttcaccat
     3241 attggccagg ctggtctcca actcctaatc tcaggtgatc tacccacctt ggcctcccaa
     3301 attgctggga ttacaggcgt gaaccactgc tcccttccct gtccttctga ttttgtaggt
     3361 aaccacgtgc ggaccgagcg gccgcaggaa cccctagtga tggagttggc cactccctct
     3421 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt
     3481 gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg
     3541 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt
     3601 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     3661 ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     3721 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     3781 gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc
     3841 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     3901 gactcttgtt ccaaactgga acaacactca actctatctc gggctattct tttgatttat
     3961 aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta
     4021 acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct
     4081 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct
     4141 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     4201 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga
     4261 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     4321 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     4381 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     4441 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     4501 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     4561 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     4621 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     4681 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     4741 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     4801 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     4861 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     4921 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     4981 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     5041 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     5101 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     5161 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     5221 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     5281 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     5341 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     5401 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     5461 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     5521 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     5581 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     5641 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     5701 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     5761 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     5821 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     5881 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     5941 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     6001 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     6061 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     6121 tgt
//
