LOCUS       sequence-495858-        6548 bp    DNA     circular SYN 19-DEC-2025
DEFINITION  Constitutive expression of WAChR-1m opsin fusion protein: 
            {WAChR-1m}-{Kir2.1 Golgi tag}-{mScarlet}-{Kv2.1 soma tag}-{Kir2.1 ER
            tag}.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6548)
  AUTHORS   Tose AJ, Nava AA, McGrath SN, Mardinly AR, Naka A
  TITLE     WAChRs are excitatory opsins sensitive to indoor lighting
  JOURNAL   bioRxiv 2025.09.12.675947; doi: 
            https://doi.org/10.1101/2025.09.12.675947
REFERENCE   2  (bases 1 to 6548)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 19, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            2025.09.12.675947; doi: https://doi.org/10.1101/2025.09.12.675947"
COMMENT     Sequence Label: pAAV-EF1a>WAChR_1m-G-mScarlet-Soma-E; WPRE3-BG...
FEATURES             Location/Qualifiers
     source          1..6548
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(398..1189,1190..1258))
                     /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     1021..1040
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1259..1363)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1431..1449
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1487..1509)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1609..1628
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(1645..2100)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1782..1803)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1994..2013
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2175..2315
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2186..2315
                     /label=AAV2 ITR
     polyA_signal    2323..2530
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2519..2536
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     primer_bind     2733..2753
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     protein_bind    2814..2838
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(3058..3750)
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet
                     /translation="VSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWDRVMNFEDGGA
                     VTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIKM
                     ALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRHS
                     TGGMDELYK"
     primer_bind     complement(3333..3352)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     3602..3620
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     protein_bind    complement(4759..4783)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        complement(4783..5961)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          4792..5730
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     complement(4815..4835)
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    complement(5962..5982)
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     repeat_region   5989..6114
                     /label=AAV2 ITR
     rep_origin      complement(join(6187..6548,1..227))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6267..6286)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
       61 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
      121 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
      181 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
      241 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
      301 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
      361 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
      421 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
      481 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcggc
      541 ttccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
      601 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
      661 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
      721 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
      781 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
      841 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
      901 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
      961 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     1021 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     1081 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     1141 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     1201 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     1261 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     1321 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     1381 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     1441 tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc
     1501 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
     1561 agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc
     1621 agattgtact gagagtgcac cataaaattg taaacgttaa tattttgtta aaattcgcgt
     1681 taaatttttg ttaaatcagc tcatttttta accaatagac cgaaatcggc aaaatccctt
     1741 ataaatcaaa agaatagccc gagatagagt tgagtgttgt tccagtttgg aacaagagtc
     1801 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     1861 gcccactacg tgaaccatca cccaaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     1921 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     1981 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc taaggcgctg gcaagtgtag
     2041 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     2101 actatggttg ctttgacgta tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
     2161 ccgcatcagg cgcccctgca ggcagctgcg cgctcgctcg ctcactgagg ccgcccgggc
     2221 aaagcccggg cgtcgggcga cctttggtcg cccggcctca gtgagcgagc gagcgcgcag
     2281 agagggagtg gccaactcca tcactagggg ttcctgcggc cctccccagc atgcctgcta
     2341 ttctcttccc aatcctcccc cttgctgtcc tgccccaccc caccccccag aatagaatga
     2401 cacctactca gacaatgcga tgcaatttcc tcattttatt aggaaaggac agtgggagtg
     2461 gcaccttcca gggtcaagga aggcacgggg gaggggcaaa caacagatgg ctggcaacta
     2521 gaaggcacag tcgaggctga tcagcgagct ctaggaattc caccacggaa ttgtcagtgc
     2581 ccaacagccg agcccctgtc cagcagcggg caaggcaggc ggcgatgagt tccgccgtgg
     2641 caagaactaa ccaggattta tacaaggagg agaaaatgaa agccatacgg gaagcaatag
     2701 catgatacaa aggcattaaa gcagcgtatc cacatagcgt aaaaggagca acatagttaa
     2761 gaataccagt caatctttca caaattttgt aatccagagg ttgattatga attcccactt
     2821 tgtacaagaa agctgggttt acacctcgtt ctcgtagcag aagaatctgg tggcttcagg
     2881 gaagtccgtg gcacagctga tgaagctgtc aatgctggac atgctccgca tgtcgatgac
     2941 gccctccgtg cgtgcgggca gaggggccac ggggctgggc atgctgctca tctccagctc
     3001 ctctggaggc ttgctctgcg gggccatctc cttggtgttg aggatgggct gggactgctt
     3061 gtacagctcg tccatgccgc cggtggagtg gcggccctcg gagcgttcgt actgttccac
     3121 cacggtgtag tcctcgttgt gggaggtgat gtccaacttg cggtccacgt tgtaggcgcc
     3181 gggcatctgc acgggcttct tggccttgta ggtggtcttg aagtccgcca ggtagcgccc
     3241 gccgtccttc aggcgcaggg ccatcttaat gtcgcccttc agcacgccgt cctcggggta
     3301 caaccgctcg gtggacgctt cccagcccat tgtcttcttc tgcattacgg ggccgtcagg
     3361 agggaagttg gtgccgcgga gcttcacctt gtagatcagg gtgccgtcct ccagggaggt
     3421 gtcctgggtc acggtcacgg cgccgccgtc ctcgaagttc atcacgcggt cccacttgaa
     3481 gccctcgggg aaggactgct tatagtagtc ggggatgtcg gcggggtgct tggtgaaggc
     3541 cctggagccg tacatgaact gaggggacag gatgtcccag gagaagggca gggggccacc
     3601 cttggtcacc ttcagcttgg cggtctgggt gccctcgtag gggcggccct cgccctcgcc
     3661 ctcgatctcg aactcgtggc cgttcatgga gccctccatg tgcaccttga accgcatgaa
     3721 ctccttgatc actgcctcgc ccttgctcac cacgttgata tcgatctggt ccagggggat
     3781 gtactcgccc tcgctggtga tcctgctctt ggcggccgct ccgctagcgt aggggtcgtc
     3841 ttcttcaacg ggcatcatca cggtggcgcc atcggcgcca gcggcggtca aggttttctg
     3901 attctcctcg tcgatctcct gtctctgcct tttcagctca tcccaggcgt agtcctcgat
     3961 gtgcagtctg cacctcagca tgaacacgcc gaaaatggtc ttgcaggtaa tgtctaggaa
     4021 ggcgtgaatg atagcaaagg cctcctcgct gatcacgccg gtggtgtgaa agcccagcag
     4081 ccacagcacg gggaacatgg gccagaagcc gaagaagatg aacatggccc tctccaggtc
     4141 cttcttggcg cagtcgggca cgtacctgat cctctttctc acctcgttga aaaagaagta
     4201 gaagaaaatg ctaaacaggg tgcagccgat agcgaagtag atgtacttct cggggctcct
     4261 cacgatgaag ctggcgaggg cgatcaccag cagcacggcg gtgctgccgg tgaaggccag
     4321 cttgtagggc acgtccacgg tgaagcacag gtccatcacg atcagggggc atgtcacgat
     4381 gtagtccagg taccagtacc tgttcaggct aaactcgctg gtgctgtccc tagcaaagtc
     4441 gctggtcttg gccacgtaca tcaggccggt taagaaggcg atggcggcag acagggtatt
     4501 cagttccagg ggtcttctct tgggatgggg gtgggctcca aagccggttc ccctccaaga
     4561 gtagtaccag aacaccacgc cgcttccaag cagaatcacg gcgctaaatc cgtcgatggt
     4621 tgcaagcagg gtgtaattca tgctcatctg aggagaaggg gcagcctgag cacaagaggt
     4681 ggtgcctccg ttaggattgg ggctttgatc tgtgggggcc accacggtag ggggtgtagg
     4741 acaggcgctc atggtggcag cctgcttttt tgtacaaact tgtcacgaca cctgaaatgg
     4801 aagaaaaaaa ctttgaacca ctgtctgagg cttgagaatg aaccaagatc caaactcaaa
     4861 aagggcaaat tccaaggaga attacatcaa gtgccaagct ggcctaactt cagtctccac
     4921 ccactcagtg tggggaaact ccatcgcata aaacccctcc ccccaaccta aagacgacgt
     4981 actccaaaag ctcgagaact aatcgaggtg cctggacggc gcccggtact ccgtggagtc
     5041 acatgaagcg acggctgagg acggaaaggc ccttttcctt tgtgtgggtg actcacccgc
     5101 ccgctctccc gagcgccgcg tcctccattt tgagctccct gcagcagggc cgggaagcgg
     5161 ccatctttcc gctcacgcaa ctggtgccga ccgggccagc cttgccgccc agggcggggc
     5221 gatacacggc ggcgcgagac caggcaccag agcaggccgg ccagcttgag actacccccg
     5281 tccgattctc ggtggccgcg ctcgcaggcc ccgcctcgcc gaacatgtgc gctgggacgc
     5341 acgggccccg tcgccgcccg cggccccaaa aaccgaaata ccagtgtgca gatcttggcc
     5401 cgcatttaca agactatctt gccagaaaaa aagcgtcgca gcaggtcatc aaaaatttta
     5461 aatggctaga gacttatcga aagcagcgag acaggcgcga aggtgccacc agattcgcac
     5521 gcggcggccc cagcgcccag gccaggcctc aactcaagca cgaggcgaag gggctcctta
     5581 agcgcaaggc ctcgaactct cccacccact tccaacccga agctcgggat caagaatcac
     5641 gtactgcagc caggtggaag taattcaagg cacgcaaggg ccataacccg taaagaggcc
     5701 aggcccgcgg gaaccacaca cggcacttac ctgtgttctg gcggcaaacc cgttgcgaaa
     5761 aagaacgttc acggcgacta ctgcacttat atacggttct cccccaccct cgggaaaaag
     5821 gcggagccag tacacgacat cactttccca gtttaccccg cgccaccttc tctaggcacc
     5881 ggttcaattg ccgacccctc cccccaactt ctcggggact gtgggcgatg tgcgctctgc
     5941 ccactgacgg gcaccggagc ccaacttttc tatacaaagt tgtctagaag gaacccctag
     6001 tgatggagtt ggccactccc tctctgcgcg ctcgctcact gaggccgggc gaccaaaggt
     6061 cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct
     6121 gcaggacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     6181 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     6241 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     6301 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     6361 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     6421 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     6481 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     6541 cactggta
//
