LOCUS       sequence-390650-        7443 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses optimized PdCO in frame with mScarlet under control of CMV
            promotor..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7443)
  AUTHORS   Wietek J, Nozownik A, Pulin M, Saraf-Sinik I, Matosevich N, 
            Gowrishankar R, Gat A, Malan D, Brown BJ, Dine J, Imambocus BN, Levy
            R, Sauter K, Litvin A, Regev N, Subramaniam S, Abrera K, Summarli D,
            Goren EM, Mizrachi G, Bitton E, Benjamin A, Copits BA, Sasse P, Rost
            BR, Schmitz D, Bruchas MR, Soba P, Oren-Suissa M, Nir Y, Wiegert JS,
            Yizhar O
  TITLE     A bistable inhibitory optoGPCR for multiplexed optogenetic control 
            of neural circuits.
  JOURNAL   Nat Methods. 2024 May 29. doi: 10.1038/s41592-024-02285-8.
  PUBMED    38811857
REFERENCE   2  (bases 1 to 7443)
  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 
            Methods. 2024 May 29. doi: 10.1038/s41592-024-02285-8."
COMMENT     Sequence Label: pcDNA3.1-CMV-PdCO-mScarlet
FEATURES             Location/Qualifiers
     source          1..7443
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        113..492
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        493..696
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     647..667
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     741..760
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        741..759
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      795..804
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1977..2672
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(2110..2128)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2378..2397
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(2743..2760)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2749..2973
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3019..3447
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3106..3125)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3316..3337
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3456..3476)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3461..3790
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3641..3776
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3703..3722
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3839..4864
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             3839..4864
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     polyA_signal    4994..5115
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5031..5050)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5085..5104
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5164..5180)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5164..5180)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5177..5199)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5188..5204
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5212..5218,5219..5236,5237..5242))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5257..5278
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5395..5412)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5566..6154)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5646..5665)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6325..7116,7117..7185))
                     /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     6948..6967
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7186..7290)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7365..7384)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt
       61 agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga
      121 ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
      181 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
      241 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
      301 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
      361 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
      421 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
      481 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
      541 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
      601 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
      661 aggcgtgtac ggtgggaggt ctatataagc agagctctct ggctaactag agaacccact
      721 gcttactggc ttatcgaaat taatacgact cactataggg agacccaagc tggctagcgt
      781 ttaaacttgg atccgccacc atggacgggg agaacttgac gattccaaat cctgtcaccg
      841 agttgatgga taccccaatc aacagcacgt atttccaaaa ccttaatgcg gaaacggacg
      901 gcggaaatca ttacatctac aatgccttca cagccaccga ttacaacatt tgtgcagctt
      961 accttttctt cattgcttgt ctgggagtca gtctcaacgt tttggtgttg gtcctcttta
     1021 tcaaagacag aaagttgagg agccccaata acttcctgta tgtaagtctc gctctcgggg
     1081 atctgttggt tgcagttttc ggcaccgcat ttaagttcat tattacagct cgcaaaaccc
     1141 tgttgcgaga agaagacggt ttctgcaaat ggtacggctt cataacatac ctcggcgggc
     1201 tcgcggcact gatgaccctg agcgttattg cttttgttcg ttgtctggcc gtgctgcgtc
     1261 tcggctcctt cacaggtttg acaacgcgga tgggcgtggc ggcgatggct ttcatttgga
     1321 tctactccct ggcattcaca ctcgcgcctc tccttgggtg gaatcactac ataccggaag
     1381 gtttggccac ttggtgtagt atcgactggc tttccgatga aacatccgat aagtcctacg
     1441 tgtttgctat tttcatcttc tgctttcttg tccctgttct cattatcgtc gtgtcatacg
     1501 ggctgatcta cgataaggtt cggaaggtag cgaaaacagg cggttctgtt gccaaagcag
     1561 aacgcgaagt actgcgcatg acgctgctga tggttagcct gttcatgctt gcctggagtc
     1621 cctacgctgt tatctgcatg ctcgctagct tcggccctaa agacctgctc catccagtgg
     1681 ccacagtgat tcctgccatg tttgcaaagt cttctacgat gtataaccct cttatctatg
     1741 tgttcatgaa caaacaattc aggcggagcc ttaaagttct tctcggaatg ggagtagagg
     1801 acctgaactc cgagagtgag agagcaactg gtggtacagc cacgaatcaa gtcgccgcga
     1861 cgaccgagac ttcccaggtt gctcctgctc cgcgagcccg agatccaacc ggtaagagca
     1921 ggatcaccag cgagggcgag tacatccccc tggaccagat cgacatcaac gtggtgatgg
     1981 tgagcaaggg cgaggcagtg atcaaggagt tcatgcggtt caaggtgcac atggagggct
     2041 ccatgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca
     2101 cccagaccgc caagctgaag gtgaccaagg gtggccccct gcccttctcc tgggacatcc
     2161 tgtcccctca gttcatgtac ggctccaggg ccttcaccaa gcaccccgcc gacatccccg
     2221 actactataa gcagtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg
     2281 acggcggcgc cgtgaccgtg acccaggaca cctccctgga ggacggcacc ctgatctaca
     2341 aggtgaagct ccgcggcacc aacttccctc ctgacggccc cgtaatgcag aagaagacaa
     2401 tgggctggga agcgtccacc gagcggttgt accccgagga cggcgtgctg aagggcgaca
     2461 ttaagatggc cctgcgcctg aaggacggcg gacgctacct ggcggacttc aagaccacct
     2521 acaaggccaa gaagcccgtg cagatgcccg gcgcctacaa cgtcgaccgc aagttggaca
     2581 tcacctccca caacgaggac tacaccgtgg tggaacagta cgaacgctcc gagggccgcc
     2641 actccaccgg cggcatggac gagctgtaca agttctgcta cgagaacgag gtgtaactag
     2701 aattcaagct ttctagaggg cccgtttaaa cccgctgatc agcctcgact gtgccttcta
     2761 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     2821 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     2881 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
     2941 gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg
     3001 gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     3061 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     3121 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     3181 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     3241 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     3301 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatcttgg
     3361 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     3421 tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg
     3481 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     3541 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     3601 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     3661 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     3721 ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     3781 cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc agcacgtgat
     3841 gaaaaagcct gaactcaccg cgacgtctgt cgagaagttt ctgatcgaaa agttcgacag
     3901 cgtctccgac ctgatgcagc tctcggaggg cgaagaatct cgtgctttca gcttcgatgt
     3961 aggagggcgt ggatatgtcc tgcgggtaaa tagctgcgcc gatggtttct acaaagatcg
     4021 ttatgtttat cggcactttg catcggccgc gctcccgatt ccggaagtgc ttgacattgg
     4081 ggaattcagc gagagcctga cctattgcat ctcccgccgt gcacagggtg tcacgttgca
     4141 agacctgcct gaaaccgaac tgcccgctgt tctgcagccg gtcgcggagg ccatggatgc
     4201 gatcgctgcg gccgatctta gccagacgag cgggttcggc ccattcggac cgcaaggaat
     4261 cggtcaatac actacatggc gtgatttcat atgcgcgatt gctgatcccc atgtgtatca
     4321 ctggcaaact gtgatggacg acaccgtcag tgcgtccgtc gcgcaggctc tcgatgagct
     4381 gatgctttgg gccgaggact gccccgaagt ccggcacctc gtgcacgcgg atttcggctc
     4441 caacaatgtc ctgacggaca atggccgcat aacagcggtc attgactgga gcgaggcgat
     4501 gttcggggat tcccaatacg aggtcgccaa catcttcttc tggaggccgt ggttggcttg
     4561 tatggagcag cagacgcgct acttcgagcg gaggcatccg gagcttgcag gatcgccgcg
     4621 gctccgggcg tatatgctcc gcattggtct tgaccaactc tatcagagct tggttgacgg
     4681 caatttcgat gatgcagctt gggcgcaggg tcgatgcgac gcaatcgtcc gatccggagc
     4741 cgggactgtc gggcgtacac aaatcgcccg cagaagcgcg gccgtctgga ccgatggctg
     4801 tgtagaagta ctcgccgata gtggaaaccg acgccccagc actcgtccga gggcaaagga
     4861 atagcacgtg ctacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     4921 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     4981 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     5041 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     5101 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
     5161 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     5221 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     5281 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     5341 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     5401 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     5461 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     5521 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     5581 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     5641 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     5701 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     5761 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     5821 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     5881 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     5941 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     6001 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     6061 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     6121 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     6181 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     6241 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     6301 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     6361 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     6421 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     6481 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     6541 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     6601 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     6661 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     6721 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     6781 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     6841 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     6901 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     6961 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     7021 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     7081 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     7141 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     7201 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     7261 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     7321 cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat ctgctctgat
     7381 gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc
     7441 gcg
//
