LOCUS       sequence-181302-        5119 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent protein reporter construct.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5119)
  AUTHORS   Wang W, Wildes CP, Pattarabanjird T, Sanchez MI, Glober GF, Matthews
            GA, Tye KM, Ting AY
  TITLE     A light- and calcium-gated transcription factor for imaging and 
            manipulating activated neurons.
  JOURNAL   Nat Biotechnol. 2017 Jun 26. doi: 10.1038/nbt.3909.
  PUBMED    28650461
REFERENCE   2  (bases 1 to 5119)
  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: "Nat 
            Biotechnol. 2017 Jun 26. doi: 10.1038/nbt.3909."
COMMENT     Sequence Label: TRE-mCherry
FEATURES             Location/Qualifiers
     source          1..5119
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   8..137
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     protein_bind    183..201
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    218..236
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    254..272
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    290..308
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    325..343
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    361..379
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        391..429
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     426..450
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      478..487
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      505..514
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             511..1221
                     /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(656..674)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     924..943
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    1253..1841
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1306..1326)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1724..1735)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    1873..2349
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2072..2091)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   2389..2529
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2389..2518
                     /label=AAV2 ITR
     rep_origin      2604..3059
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2691..2710)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2901..2922
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3076..3095)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3195..3217
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3255..3273)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3341..3445
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3446..3514,3515..4306)
                     /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(3664..3683)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4477..5065
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4966..4985
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cacatgtcct gcaggcagct gcgcgctcgc tcgctcactg aggccgcccg ggcgtcgggc
       61 gacctttggt cgcccggcct cagtgagcga gcgagcgcgc agagagggag tggccaactc
      121 catcactagg ggttcctgcg gccgcacgcg tccttcaggt ccgaggttct agacgagttt
      181 actccctatc agtgatagag aacgatgtcg agtttactcc ctatcagtga tagagaacgt
      241 atgtcgagtt tactccctat cagtgataga gaacgtatgt cgagtttact ccctatcagt
      301 gatagagaac gtatgtcgag tttatcccta tcagtgatag agaacgtatg tcgagtttac
      361 tccctatcag tgatagagaa cgtatgtcga ggtaggcgtg tacggtggga ggcctatata
      421 agcagagctc gtttagtgaa ccgtcagatc gcaaagggcg aattcgatcc accggtcgcc
      481 accatggctc gggatccacc ggtcgccacc atggtgagca agggcgagga ggataacatg
      541 gccatcatca aggagttcat gcgcttcaag gtgcacatgg agggctccgt gaacggccac
      601 gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag
      661 ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc ccctcagttc
      721 atgtacggct ccaaggccta cgtgaagcac cccgccgaca tccccgacta cttgaagctg
      781 tccttccccg agggattcaa gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg
      841 accgtgaccc aggactcctc cctgcaggac ggcgagttca tctacaaggt gaagctgcgc
      901 ggcaccaact tcccctccga cggccccgta atgcagaaga agaccatggg ctgggaggcc
      961 tcctccgagc ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa gcagaggctg
     1021 aagctgaagg acggcggcca ctacgacgct gaggtcaaga ccacctacaa ggccaagaag
     1081 cccgtgcagc tgcccggcgc ctacaacgtc aacatcaagt tggacatcac ctcccacaac
     1141 gaggactaca ccatcgtgga acagtacgaa cgcgccgagg gccgccactc caccggcggc
     1201 atggacgagc tgtacaagta aactagtacc ggtagatatc aagcttatcg ataatcaacc
     1261 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     1321 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     1381 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     1441 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     1501 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     1561 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     1621 tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctatgt
     1681 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     1741 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     1801 ccctcagacg agtcggatct ccctttgggc cgcctccccg catcgatacc gagcgctgct
     1861 cgagagatct acgggtggca tccctgtgac ccctccccag tgcctctcct ggccctggaa
     1921 gttgccactc cagtgcccac cagccttgtc ctaataaaat taagttgcat cattttgtct
     1981 gactaggtgt ccttctataa tattatgggg tggagggggg tggtatggag caaggggcaa
     2041 gttgggaaga caacctgtag ggcctgcggg gtctattggg aaccaagctg gagtgcagtg
     2101 gcacaatctt ggctcactgc aatctccgcc tcctgggttc aagcgattct cctgcctcag
     2161 cctcccgagt tgttgggatt ccaggcatgc atgaccaggc tcagctaatt tttgtttttt
     2221 tggtagagac ggggtttcac catattggcc aggctggtct ccaactccta atctcaggtg
     2281 atctacccac cttggcctcc caaattgctg ggattacagg cgtgaaccac tgctcccttc
     2341 cctgtccttc tgattttgta ggtaaccacg tgcggaccga gcggccgcag gaacccctag
     2401 tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa
     2461 aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct
     2521 gcctgcaggg gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     2581 gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     2641 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc
     2701 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     2761 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt
     2821 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     2881 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaactctat
     2941 ctcgggctat tcttttgatt tataagggat tttgccgatt tcggtctatt ggttaaaaaa
     3001 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt
     3061 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     3121 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     3181 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     3241 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     3301 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     3361 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     3421 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     3481 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     3541 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     3601 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     3661 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     3721 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     3781 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     3841 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     3901 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     3961 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     4021 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     4081 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     4141 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     4201 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     4261 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     4321 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     4381 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     4441 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     4501 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     4561 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     4621 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     4681 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     4741 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     4801 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     4861 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     4921 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     4981 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     5041 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     5101 cttttgctgg ccttttgct
//
