LOCUS       sequence-495874-        6524 bp    DNA     circular SYN 19-DEC-2025
DEFINITION  Constitutive expression of WAChR-1s opsin fusion protein: 
            {WAChR-1s}-{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 6524)
  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 6524)
  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_1s-G-mScarlet-Soma-E; WPRE3-BG...
FEATURES             Location/Qualifiers
     source          1..6524
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    115..135
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     promoter        136..1314
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          367..1305
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1262..1282
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    1314..1338
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2477..2495)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     protein_bind    complement(3253..3277)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(3338..3358)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     primer_bind     complement(3555..3572)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3561..3768
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3776..3916
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3776..3905
                     /label=AAV2 ITR
     rep_origin      3991..4446
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4078..4097)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4288..4309
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4463..4482)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4582..4604
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4642..4660)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4728..4832
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4833..4901,4902..5693)
                     /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(5051..5070)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5864..6452
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6353..6372
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcggcc tcagtgagcg
       61 agcgagcgcg cagagaggga gtggccaact ccatcactag gggttccttc tagacaactt
      121 tgtatagaaa agttgggctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc
      181 cccgagaagt tggggggagg ggtcggcaat tgaaccggtg cctagagaag gtggcgcggg
      241 gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa
      301 ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga
      361 acacaggtaa gtgccgtgtg tggttcccgc gggcctggcc tctttacggg ttatggccct
      421 tgcgtgcctt gaattacttc cacctggctg cagtacgtga ttcttgatcc cgagcttcgg
      481 gttggaagtg ggtgggagag ttcgaggcct tgcgcttaag gagccccttc gcctcgtgct
      541 tgagttgagg cctggcctgg gcgctggggc cgccgcgtgc gaatctggtg gcaccttcgc
      601 gcctgtctcg ctgctttcga taagtctcta gccatttaaa atttttgatg acctgctgcg
      661 acgctttttt tctggcaaga tagtcttgta aatgcgggcc aagatctgca cactggtatt
      721 tcggtttttg gggccgcggg cggcgacggg gcccgtgcgt cccagcgcac atgttcggcg
      781 aggcggggcc tgcgagcgcg gccaccgaga atcggacggg ggtagtctca agctggccgg
      841 cctgctctgg tgcctggtct cgcgccgccg tgtatcgccc cgccctgggc ggcaaggctg
      901 gcccggtcgg caccagttgc gtgagcggaa agatggccgc ttcccggccc tgctgcaggg
      961 agctcaaaat ggaggacgcg gcgctcggga gagcgggcgg gtgagtcacc cacacaaagg
     1021 aaaagggcct ttccgtcctc agccgtcgct tcatgtgact ccacggagta ccgggcgccg
     1081 tccaggcacc tcgattagtt ctcgagcttt tggagtacgt cgtctttagg ttggggggag
     1141 gggttttatg cgatggagtt tccccacact gagtgggtgg agactgaagt taggccagct
     1201 tggcacttga tgtaattctc cttggaattt gccctttttg agtttggatc ttggttcatt
     1261 ctcaagcctc agacagtggt tcaaagtttt tttcttccat ttcaggtgtc gtgacaagtt
     1321 tgtacaaaaa agcaggctgc caccatgtct gcctgcccca caccccctac agtggtggcc
     1381 cccacagacc agagcccaaa ccctaatgga ggcaccacca gctgtgccca ggctgccccc
     1441 agccctcaga tgagcatgaa ctacaccctg ttggccacca ttgatggctt ctctgctgtg
     1501 attctgctgg gcagtggggt ggtgttctgg tactacagct ggaggggcac aggctttgga
     1561 gcccatcccc accctaagag gaggcccctg gagctgaaca ccctgtctgc tgccattgct
     1621 ttcttaacgg gattgatgta tgtggccaag acctctgact ttgccagaga cagcacctca
     1681 gagttcagcc tgaacagata ctggtactct gactacattg tgacctgccc cctgatagtg
     1741 atggacctgt gcttcacagt ggatgtgccc tacaagctgg ccttcacagg cagcacagct
     1801 gtcctgctgg tgattgccct ggccagcttc attgtgagaa gccctgagaa gtacatctat
     1861 tttgccattg gctgcaccct gttcagcatc ttcttctact tcttctttaa tgaggtgaga
     1921 aagagaatca ggtatgtgcc tgactgtgcc aagaaggacc tggagagggc catgtggatc
     1981 ttctttggct tctggcccat gttcccagtc ctgtggctgc tgggcttcca caccactgga
     2041 gtgatctctg aggaggcctt tgccatcatc catgccttct tagacattac ctgcaagacc
     2101 atctttgggg tgttcatgct gagatgcaga ctgcacattg aggactatgc ctgggatgag
     2161 ctgaagaggc agagacagga gattgatgag gagaaccaga agaccctgac agctgctggg
     2221 gctgatggag ccactgtgat gatgcctgtg gaggaggatg acccctatgc ctctggagct
     2281 gctgccaaga gcaggatcac ctctgaggga gagtacatcc ccctggacca gattgacatc
     2341 aatgtgatgg acagcacaga ggctgtgatc aaggagttca tgaggttcaa ggtgcacatg
     2401 gagggcagca tgaatggcca tgagtttgag attgagggag aaggagaggg caggccctat
     2461 gagggcaccc agacagccaa gctgaaggtg accaagggag gccccctgcc cttcagctgg
     2521 gacatcctga gcccacagtt catgtatggc tccagagcct tcatcaagca ccctgctgac
     2581 atccctgact actggaagca gagcttccct gagggcttca agtgggagag ggtgatgatc
     2641 tttgaggatg ggggcactgt gtctgtgacc caggacacca gcctggagga tggcaccctg
     2701 atctacaagg tgaagctgag gggaggcaac ttccctcctg atggcccagt gatgcagaag
     2761 aggaccatgg gctgggaggc ctccacagag aggctgtacc ctgaggatgt ggtgctgaag
     2821 ggggacatca agatggccct gaggctgaag gatggaggca gatacctggc tgacttcaag
     2881 accacctaca aggccaagaa acctgtgcag atgcctgggg ccttcaacat tgacagaaag
     2941 ctggacatca ctagccacaa tgaggactac acagttgtgg agcagtatga gagatctgtg
     3001 gccaggcaca gcactggagg ctctgggggc tcccagagcc agcccatcct gaacaccaag
     3061 gagatggccc ctcagagcaa gccccctgag gagctggaga tgagcagcat gcccagccct
     3121 gtggctcccc tgcctgccag aacagagggg gtgattgaca tgagaagcat gagctccatt
     3181 gacagcttca tctcctgtgc cacagacttc ccagaggcca ccagattctt ctgctatgaa
     3241 aatgaggttt gaacccagct ttcttgtaca aagtgggaat tcataatcaa cctctggatt
     3301 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     3361 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
     3421 cctccttgta taaatcctgg ttagttcttg ccacggcgga actcatcgcc gcctgccttg
     3481 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg gaattcctag
     3541 agctcgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     3601 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     3661 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     3721 ggacagcaag ggggaggatt gggaagagaa tagcaggcat gctggggagg gccgcaggaa
     3781 cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg
     3841 cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg
     3901 cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     3961 tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg
     4021 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc
     4081 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     4141 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     4201 ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     4261 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     4321 actctatctc gggctattct tttgatttat aagggatttt gccgatttcg gtctattggt
     4381 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta
     4441 caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc
     4501 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     4561 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     4621 cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     4681 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     4741 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     4801 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     4861 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     4921 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     4981 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     5041 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     5101 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     5161 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     5221 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     5281 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     5341 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     5401 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     5461 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     5521 ctgataaatc tggagccggt gagcgtggaa gccgcggtat cattgcagca ctggggccag
     5581 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     5641 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     5701 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     5761 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     5821 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     5881 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     5941 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     6001 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     6061 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     6121 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     6181 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     6241 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     6301 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     6361 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     6421 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     6481 tcctggcctt ttgctggcct tttgctcaca tgtcctgcag gcag
//
