LOCUS       sequence-412557-        5823 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 5823)
  TITLE     Prigge Lab AAV vectors
REFERENCE   2  (bases 1 to 5823)
  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_CKII(0.4)_AzPPO_D14_ts_mScarlet_Kv2.1
FEATURES             Location/Qualifiers
     source          1..5823
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      528..537
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1689..2384
                     /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(1822..1840)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2090..2109
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    2598..3001
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     polyA_signal    3008..3056
                     /note="synthetic polyadenylation signal"
     repeat_region   3086..3226
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3086..3215
                     /label=AAV2 ITR
     rep_origin      3301..3756
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3388..3407)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3598..3619
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3773..3792)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3892..3914
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3952..3970)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4038..4142
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4143..4211,4212..5003)
                     /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(4361..4380)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5174..5762
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5663..5682
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtcttgtg gactaagttt gttcgcatcc ccttctccaa
      181 ccccctcagt acatcaccct gggggaacag ggtccacttg ctcctgggcc cacacagtcc
      241 tgcagtattg tgtatataag gccagggcaa agaggagcag gttttaaagt gaaaggcagg
      301 caggtgttgg ggaggcagtt accggggcaa cgggaacagg gcgtttcgga ggtggttgcc
      361 atggggacct ggatgctgac gaaggctcgc gaggctgtga gcagccacag tgccctgctc
      421 agaagcccca agctcgtcag tcaagccggt tctccgtttg cactcaggag cacgggcagg
      481 cgagtggccc ctagttctgg gggcagctct agagcggtac cggatccgcc accatggaga
      541 gtgttgcctt ccaccggaac tcttcatcat ctaatagcag cgcctacacg gagctcttgt
      601 ctcggactgg ttacacaatc ctggccatta tcatgggcgc gttctctgta ggagggatca
      661 tactcaatgt cttggtaatt gtggtgacag tgagacacag acagcttagg cagcccctca
      721 gctatgccct cgttaacctg gcagtttgcg acctgggctg cgctgtgttc gggggccttc
      781 ccacgacagt aaccagtgcc atgggatact tcagcctggg aagggtgggc tgtgtgttag
      841 aagccttcgc tgtttccttc ttcggtatag caagtctgtg tacagtagca gtcatctctg
      901 tggaacgcta cattgtggtg tgctatccca tgggtgcagt cctgttccag accaggcatg
      961 ccgttggcgg ggtggtcctg tcctgggtgt ggtcgtttgt gtggagcact ccacctctgt
     1021 ttgggtgggg aacgtttgag ttggagggcg tccaaaccag ttgtgctcct aactggtata
     1081 gccgcgatat tggggacatg tcctacatga tcatgttctt tgtcctttgc tttgccgtgc
     1141 ccttttccat catcatggtg tcctactcac gacttctctg gaccctccgc caggtgacca
     1201 agctgcaggt gtccgaaaca ggtagcacaa atcgtgtgga ggtgcaggtg gcacggatga
     1261 tagtggtgat ggtgttggcc ttcctagtca cctggctgcc atatgcctca atggctgtcg
     1321 ctgtcatcat ggattccagt ctatatgtag accctgtcat cgcgaccatc cctgtatact
     1381 ttgcaaaaag tagcactgtc tacaacccga tcatttatat tttcatgaac agacagtttc
     1441 gaggctacac tgttgctgct gttctgtgtg gatggaaccc atggtcctca gagccacaaa
     1501 cttctgagaa tgagaccact gttacatctg tcagcaagac cccccaaaag attgtgcctg
     1561 aaaaatcgtt agaaaccgag accagccagg tggctcctgc tccgcgagcc cgagatccaa
     1621 ccggtaagag caggatcacc agcgagggcg agtacatccc cctggaccag atcgacatca
     1681 acgtggtgat ggtgagcaag ggcgaggcag tgatcaagga gttcatgcgg ttcaaggtgc
     1741 acatggaggg ctccatgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc
     1801 cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttct
     1861 cctgggacat cctgtcccct cagttcatgt acggctccag ggccttcacc aagcaccccg
     1921 ccgacatccc cgactactat aagcagtcct tccccgaggg cttcaagtgg gagcgcgtga
     1981 tgaacttcga ggacggcggc gccgtgaccg tgacccagga cacctccctg gaggacggca
     2041 ccctgatcta caaggtgaag ctccgcggca ccaacttccc tcctgacggc cccgtaatgc
     2101 agaagaagac aatgggctgg gaagcgtcca ccgagcggtt gtaccccgag gacggcgtgc
     2161 tgaagggcga cattaagatg gccctgcgcc tgaaggacgg cggacgctac ctggcggact
     2221 tcaagaccac ctacaaggcc aagaagcccg tgcagatgcc cggcgcctac aacgtcgacc
     2281 gcaagttgga catcacctcc cacaacgagg actacaccgt ggtggaacag tacgaacgct
     2341 ccgagggccg ccactccacc ggcggcatgg acgagctgta caagcagtcc cagcctattc
     2401 tgaacaccaa agagatggcc ccccagagta aaccccctga ggagctggaa atgtcctcaa
     2461 tgccctcccc tgtggcacct cttcctgcaa gaacagaggg cgtgatcgac atgaggtcca
     2521 tgagttctat tgactctttt atctcatgcg ccaccgattt tccagaggct acgcggttta
     2581 agtaagaatt caagcttaat caacctctgg attacaaaat ttgtgaaaga ttgactgata
     2641 ttcttaacta tgttgctcct tttacgctgt gtggatatgc tgctttaatg cctctgtatc
     2701 atgctattgc ttcccgtatg gctttcgttt tctcctcctt gtataaatcc tggttgctgt
     2761 ctctttatga ggagttgtgg cccgttgtcc gtcaacgtgg cgtggtgtgc tctgtgtttg
     2821 ctgacgcaac ccccactggc tggggcattg ccaccacctg tcaactcctt tctgggactt
     2881 tcgctttccc cctcccgatc gccacggcag aactcatcgc cgcctgcctt gcccgctgct
     2941 ggacaggggc taggttgctg ggcactgata attccgtggt gttgtcgggg aagctgacgt
     3001 cagcgctaat aaaatcgcta tccatcgaag atggatgtgt gttggttttt tgtgtgggta
     3061 acccacgtgc ggaccgagcg gccgcaggaa cccctagtga tggagttggc cactccctct
     3121 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt
     3181 gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg
     3241 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt
     3301 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     3361 ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     3421 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     3481 gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc
     3541 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     3601 gactcttgtt ccaaactgga acaacactca actctatctc gggctattct tttgatttat
     3661 aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta
     3721 acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct
     3781 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct
     3841 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     3901 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga
     3961 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     4021 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     4081 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     4141 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     4201 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     4261 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     4321 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     4381 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     4441 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     4501 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     4561 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     4621 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     4681 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     4741 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     4801 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     4861 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     4921 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     4981 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     5041 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     5101 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     5161 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     5221 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     5281 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     5341 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     5401 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     5461 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     5521 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     5581 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     5641 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttt
     5701 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     5761 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     5821 tgt
//
