LOCUS       sequence-412575-        6304 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bistable opsin for neuronal inhibition.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6304)
  TITLE     Prigge Lab AAV vectors
REFERENCE   2  (bases 1 to 6304)
  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"
COMMENT     Sequence Label: pAAV_CKI(0.4)_HA_AzPPO_AgeI_nrxn1a_WT_IRES_mSc...
FEATURES             Location/Qualifiers
     source          1..6304
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   30..159
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     CDS             566..592
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             596..622
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    1840..2391
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1984..2001)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2211..2230
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2394..3089
                     /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(2527..2545)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2795..2814
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    3108..3511
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     polyA_signal    3518..3566
                     /note="synthetic polyadenylation signal"
     repeat_region   3596..3736
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3596..3725
                     /label=AAV2 ITR
     rep_origin      3811..4266
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3898..3917)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4108..4129
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4283..4302)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4402..4424
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4462..4480)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4548..4652
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4653..4721,4722..5513)
                     /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(4871..4890)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5684..6272
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6173..6192
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggccttttgc tggccttttg ctcacatgtc ctgcaggcag ctgcgcgctc gctcgctcac
       61 tgaggccgcc cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc
      121 gcagagaggg agtggccaac tccatcacta ggggttcctg cggccgcacg cgtcttgtgg
      181 actaagtttg ttcgcatccc cttctccaac cccctcagta catcaccctg ggggaacagg
      241 gtccacttgc tcctgggccc acacagtcct gcagtattgt gtatataagg ccagggcaaa
      301 gaggagcagg ttttaaagtg aaaggcaggc aggtgttggg gaggcagtta ccggggcaac
      361 gggaacaggg cgtttcggag gtggttgcca tggggacctg gatgctgacg aaggctcgcg
      421 aggctgtgag cagccacagt gccctgctca gaagccccaa gctcgtcagt caagccggtt
      481 ctccgtttgc actcaggagc acgggcaggc gagtggcccc tagttctggg ggcagctcta
      541 gagcggtacc ggatccgcca ccatgtaccc atacgatgtt ccagattacg ctggttaccc
      601 ttacgacgtg cctgactacg ccgagagtgt tgccttccac cggaactctt catcatctaa
      661 tagcagcgcc tacacggagc tcttgtctcg gactggttac acaatcctgg ccattatcat
      721 gggcgcgttc tctgtaggag ggatcatact caatgtcttg gtaattgtgg tgacagtgag
      781 acacagacag cttaggcagc ccctcagcta tgccctcgtt aacctggcag tttgcgacct
      841 gggctgcgct gtgttcgggg gccttcccac gacagtaacc agtgccatgg gatacttcag
      901 cctgggaagg gtgggctgtg tgttagaagc cttcgctgtt tccttcttcg gtatagcaag
      961 tctgtgtaca gtagcagtca tctctgtgga acgctacatt gtggtgtgct atcccatggg
     1021 tgcagtcctg ttccagacca ggcatgccgt tggcggggtg gtcctgtcct gggtgtggtc
     1081 gtttgtgtgg agcactccac ctctgtttgg gtggggaacg tttgagttgg agggcgtcca
     1141 aaccagttgt gctcctaact ggtatagccg cgatattggg gacatgtcct acatgatcat
     1201 gttctttgtc ctttgctttg ccgtgccctt ttccatcatc atggtgtcct actcacgact
     1261 tctctggacc ctccgccagg tgaccaagct gcaggtgtcc gaaacaggta gcacaaatcg
     1321 tgtggaggtg caggtggcac ggatgatagt ggtgatggtg ttggccttcc tagtcacctg
     1381 gctgccatat gcctcaatgg ctgtcgctgt catcatggat tccagtctat atgtagaccc
     1441 tgtcatcgcg accatccctg tatactttgc aaaaagtagc actgtctaca acccgatcat
     1501 ttatattttc atgaacagac agtttcgagg ctacactgtt gctgctgttc tgtgtggatg
     1561 gaacccatgg tcctcagagc cacaaacttc tgagaatgag accactgtta catctgtcag
     1621 caagaccccc caaaagattg tgcctgaaaa atcgttagaa accggtaagt accggaatag
     1681 ggatgagggg tcctaccacg tggacgaatc gcggaactac attagcaact ccgcccagtc
     1741 aaacggagcc gtggtcaagg aaaagcagcc aagctccgcc aagagcgcca acaagaacaa
     1801 gaagaacaag gacaaggagt actatgtgtg acccccctaa cgttactggc cgaagccgct
     1861 tggaataagg ccggtgtgcg tttgtctata tgttattttc caccatattg ccgtcttttg
     1921 gcaatgtgag ggcccggaaa cctggccctg tcttcttgac gagcattcct aggggtcttt
     1981 cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg
     2041 aagcttcttg aagacaaaca acgtctgtag cgaccctttg caggcagcgg aaccccccac
     2101 ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata agatacacct gcaaaggcgg
     2161 cacaacccca gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa tggctctcct
     2221 caagcgtatt caacaagggg ctgaaggatg cccagaaggt accccattgt atgggatctg
     2281 atctggggcc tcggtgcaca tgctttacat gtgtttagtc gaggttaaaa aacgtctagg
     2341 ccccccgaac cacggggacg tggttttcct ttgaaaaaca cgatgataat atgatggtga
     2401 gcaagggcga ggcagtgatc aaggagttca tgcggttcaa ggtgcacatg gagggctcca
     2461 tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccctac gagggcaccc
     2521 agaccgccaa gctgaaggtg accaagggtg gccccctgcc cttctcctgg gacatcctgt
     2581 cccctcagtt catgtacggc tccagggcct tcaccaagca ccccgccgac atccccgact
     2641 actataagca gtccttcccc gagggcttca agtgggagcg cgtgatgaac ttcgaggacg
     2701 gcggcgccgt gaccgtgacc caggacacct ccctggagga cggcaccctg atctacaagg
     2761 tgaagctccg cggcaccaac ttccctcctg acggccccgt aatgcagaag aagacaatgg
     2821 gctgggaagc gtccaccgag cggttgtacc ccgaggacgg cgtgctgaag ggcgacatta
     2881 agatggccct gcgcctgaag gacggcggac gctacctggc ggacttcaag accacctaca
     2941 aggccaagaa gcccgtgcag atgcccggcg cctacaacgt cgaccgcaag ttggacatca
     3001 cctcccacaa cgaggactac accgtggtgg aacagtacga acgctccgag ggccgccact
     3061 ccaccggcgg catggacgag ctgtacaagt agctagaatt caagcttaat caacctctgg
     3121 attacaaaat ttgtgaaaga ttgactgata ttcttaacta tgttgctcct tttacgctgt
     3181 gtggatatgc tgctttaatg cctctgtatc atgctattgc ttcccgtatg gctttcgttt
     3241 tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtcc
     3301 gtcaacgtgg cgtggtgtgc tctgtgtttg ctgacgcaac ccccactggc tggggcattg
     3361 ccaccacctg tcaactcctt tctgggactt tcgctttccc cctcccgatc gccacggcag
     3421 aactcatcgc cgcctgcctt gcccgctgct ggacaggggc taggttgctg ggcactgata
     3481 attccgtggt gttgtcgggg aagctgacgt cagcgctaat aaaatcgcta tccatcgaag
     3541 atggatgtgt gttggttttt tgtgtgggta acccacgtgc ggaccgagcg gccgcaggaa
     3601 cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg
     3661 cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg
     3721 cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     3781 tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg
     3841 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc
     3901 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     3961 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     4021 ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     4081 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     4141 actctatctc gggctattct tttgatttat aagggatttt gccgatttcg gtctattggt
     4201 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta
     4261 caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc
     4321 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     4381 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     4441 cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     4501 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     4561 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     4621 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     4681 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     4741 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     4801 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     4861 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     4921 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     4981 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     5041 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     5101 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     5161 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     5221 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     5281 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     5341 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     5401 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     5461 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     5521 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     5581 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     5641 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     5701 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     5761 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     5821 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     5881 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     5941 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     6001 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     6061 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     6121 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     6181 cctggtatct ttatagtcct gtcgggtttt gccacctctg acttgagcgt cgatttttgt
     6241 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     6301 tcct
//
