LOCUS       sequence-390696-        5670 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses optimized PdCO in frame with mScarlet under control of 
            minimal CamKIIa promotor..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5670)
  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 5670)
  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: pAAV-CaMKIIa(0.4)-PdCO-mScarlet-WPRE
FEATURES             Location/Qualifiers
     source          1..5670
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     32..51
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   193..322
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      720..729
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1902..2597
                     /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(2035..2053)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2303..2322
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    2637..3040
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     polyA_signal    3047..3095
                     /note="synthetic polyadenylation signal"
     repeat_region   3125..3265
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3125..3254
                     /label=AAV2 ITR
     rep_origin      3340..3795
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3427..3446)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3637..3658
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3812..3831)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3931..3953
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3991..4009)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4077..4181
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4182..4250,4251..5042)
                     /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(4400..4419)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(5213..5670,1..131)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
       61 tcgggttttg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
      121 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
      181 ttgctcacat gtcctgcagg cagctgcgcg ctcgctcgct cactgaggcc gcccgggcgt
      241 cgggcgacct ttggtcgccc ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc
      301 aactccatca ctaggggttc ctgcggccgc acgcgtcttg tggactaagt ttgttcgcat
      361 ccccttctcc aaccccctca gtacatcacc ctgggggaac agggtccact tgctcctggg
      421 cccacacagt cctgcagtat tgtgtatata aggccagggc aaagaggagc aggttttaaa
      481 gtgaaaggca ggcaggtgtt ggggaggcag ttaccggggc aacgggaaca gggcgtttcg
      541 gaggtggttg ccatggggac ctggatgctg acgaaggctc gcgaggctgt gagcagccac
      601 agtgccctgc tcagaagccc caagctcgtc agtcaagccg gttctccgtt tgcactcagg
      661 agcacgggca ggcgagtggc ccctagttct gggggcagct ctagagcggt accggatccg
      721 ccaccatgga cggggagaac ttgacgattc caaatcctgt caccgagttg atggataccc
      781 caatcaacag cacgtatttc caaaacctta atgcggaaac ggacggcgga aatcattaca
      841 tctacaatgc cttcacagcc accgattaca acatttgtgc agcttacctt ttcttcattg
      901 cttgtctggg agtcagtctc aacgttttgg tgttggtcct ctttatcaaa gacagaaagt
      961 tgaggagccc caataacttc ctgtatgtaa gtctcgctct cggggatctg ttggttgcag
     1021 ttttcggcac cgcatttaag ttcattatta cagctcgcaa aaccctgttg cgagaagaag
     1081 acggtttctg caaatggtac ggcttcataa catacctcgg cgggctcgcg gcactgatga
     1141 ccctgagcgt tattgctttt gttcgttgtc tggccgtgct gcgtctcggc tccttcacag
     1201 gtttgacaac gcggatgggc gtggcggcga tggctttcat ttggatctac tccctggcat
     1261 tcacactcgc gcctctcctt gggtggaatc actacatacc ggaaggtttg gccacttggt
     1321 gtagtatcga ctggctttcc gatgaaacat ccgataagtc ctacgtgttt gctattttca
     1381 tcttctgctt tcttgtccct gttctcatta tcgtcgtgtc atacgggctg atctacgata
     1441 aggttcggaa ggtagcgaaa acaggcggtt ctgttgccaa agcagaacgc gaagtactgc
     1501 gcatgacgct gctgatggtt agcctgttca tgcttgcctg gagtccctac gctgttatct
     1561 gcatgctcgc tagcttcggc cctaaagacc tgctccatcc agtggccaca gtgattcctg
     1621 ccatgtttgc aaagtcttct acgatgtata accctcttat ctatgtgttc atgaacaaac
     1681 aattcaggcg gagccttaaa gttcttctcg gaatgggagt agaggacctg aactccgaga
     1741 gtgagagagc aactggtggt acagccacga atcaagtcgc cgcgacgacc gagacttccc
     1801 aggttgctcc tgctccgcga gcccgagatc caaccggtaa gagcaggatc accagcgagg
     1861 gcgagtacat ccccctggac cagatcgaca tcaacgtggt gatggtgagc aagggcgagg
     1921 cagtgatcaa ggagttcatg cggttcaagg tgcacatgga gggctccatg aacggccacg
     1981 agttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag accgccaagc
     2041 tgaaggtgac caagggtggc cccctgccct tctcctggga catcctgtcc cctcagttca
     2101 tgtacggctc cagggccttc accaagcacc ccgccgacat ccccgactac tataagcagt
     2161 ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc ggcgccgtga
     2221 ccgtgaccca ggacacctcc ctggaggacg gcaccctgat ctacaaggtg aagctccgcg
     2281 gcaccaactt ccctcctgac ggccccgtaa tgcagaagaa gacaatgggc tgggaagcgt
     2341 ccaccgagcg gttgtacccc gaggacggcg tgctgaaggg cgacattaag atggccctgc
     2401 gcctgaagga cggcggacgc tacctggcgg acttcaagac cacctacaag gccaagaagc
     2461 ccgtgcagat gcccggcgcc tacaacgtcg accgcaagtt ggacatcacc tcccacaacg
     2521 aggactacac cgtggtggaa cagtacgaac gctccgaggg ccgccactcc accggcggca
     2581 tggacgagct gtacaagttc tgctacgaga acgaggtgta actagaattc aagcttaatc
     2641 aacctctgga ttacaaaatt tgtgaaagat tgactgatat tcttaactat gttgctcctt
     2701 ttacgctgtg tggatatgct gctttaatgc ctctgtatca tgctattgct tcccgtatgg
     2761 ctttcgtttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc
     2821 ccgttgtccg tcaacgtggc gtggtgtgct ctgtgtttgc tgacgcaacc cccactggct
     2881 ggggcattgc caccacctgt caactccttt ctgggacttt cgctttcccc ctcccgatcg
     2941 ccacggcaga actcatcgcc gcctgccttg cccgctgctg gacaggggct aggttgctgg
     3001 gcactgataa ttccgtggtg ttgtcgggga agctgacgtc agcgctaata aaatcgctat
     3061 ccatcgaaga tggatgtgtg ttggtttttt gtgtgggtaa cccacgtgcg gaccgagcgg
     3121 ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact
     3181 gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc
     3241 gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt attttctcct tacgcatctg
     3301 tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat
     3361 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccttag
     3421 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     3481 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     3541 ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     3601 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     3661 caacactcaa ctctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg
     3721 tctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     3781 taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     3841 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     3901 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     3961 cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta
     4021 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     4081 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     4141 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     4201 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     4261 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     4321 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     4381 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     4441 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     4501 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     4561 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     4621 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     4681 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     4741 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     4801 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     4861 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     4921 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     4981 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     5041 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     5101 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     5161 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     5221 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     5281 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     5341 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     5401 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     5461 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     5521 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     5581 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     5641 cggacaggta tccggtaagc ggcagggtcg
//
