LOCUS       sequence-204350-        7200 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses a soma-targeted GtACR2 in mammalian neurons under the 
            CaMKIIa promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7200)
  AUTHORS   Mahn M, Gibor L, Patil P, Cohen-Kashi Malina K, Oring S, Printz Y, 
            Levy R, Lampl I, Yizhar O
  TITLE     High-efficiency optogenetic silencing with soma-targeted 
            anion-conducting channelrhodopsins.
  JOURNAL   Nat Commun. 2018 Oct 8;9(1):4125. doi: 10.1038/s41467-018-06511-8.
  PUBMED    30297821
REFERENCE   2  (bases 1 to 7200)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2018-10-8"; volume: "9(1):4125. doi"; pages: " 
            10.1038/s41467-018-06511-8"
COMMENT     Sequence Label: pAAV-CKIIa-stGtACR2-FusionRed
FEATURES             Location/Qualifiers
     source          1..7200
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        149..1437
                     /label=CaMKII promoter
                     /note="promoter for the mouse calcium/calmodulin-dependent 
                     kinase II alpha subunit"
     regulatory      1458..1467
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2406..3101
                     /codon_start=1
                     /product="monomeric red fluorescent protein optimized for 
                     use as a fusion tag (Shemiakina et al., 2012)"
                     /label=FusionRed
                     /note="mammalian codon-optimized"
                     /translation="MVSELIKENMPMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK
                     VVEGGPLPFAFDILATSFMYGSRTFIKHPPGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKVRGVNFPANGPVMQKKTLGWEASTETMYPADGGLEGACDMAL
                     KLVGGGHLICNLETTYRSKKPATNLKMPGVYNVDHRLERIKEADDETYVEQHEVAVARY
                     STGGAGDGGK"
     misc_feature    3327..3915
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3380..3400)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3798..3809)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3947..4423
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4146..4165)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4463..4603
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4463..4592
                     /label=AAV2 ITR
     rep_origin      4678..5133
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4765..4784)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4975..4996
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5150..5169)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5269..5291
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5329..5347)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5415..5519
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5520..5588,5589..6380)
                     /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(5738..5757)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6551..7139
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7040..7059
                     /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 gcgtttaaca ttatggcctt aggtcacttc atctccatgg
      181 ggttcttctt ctgattttct agaaaatgag atgggggtgc agagagcttc ctcagtgacc
      241 tgcccagggt cacatcagaa atgtcagagc tagaacttga actcagatta ctaatcttaa
      301 attccatgcc ttgggggcat gcaagtacga tatacagaag gagtgaactc attagggcag
      361 atgaccaatg agtttaggaa agaagagtcc agggcagggt acatctacac cacccgccca
      421 gccctgggtg agtccagcca cgttcacctc attatagttg cctctctcca gtcctacctt
      481 gacgggaagc acaagcagaa actgggacag gagccccagg agaccaaatc ttcatggtcc
      541 ctctgggagg atgggtgggg agagctgtgg cagaggcctc aggaggggcc ctgctgctca
      601 gtggtgacag ataggggtga gaaagcagac agagtcattc cgtcagcatt ctgggtctgt
      661 ttggtacttc ttctcacgct aaggtggcgg tgtgatatgc acaatggcta aaaagcaggg
      721 agagctggaa agaaacaagg acagagacag aggccaagtc aaccagacca attcccagag
      781 gaagcaaaga aaccattaca gagactacaa gggggaaggg aaggagagat gaattagctt
      841 cccctgtaaa ccttagaacc cagctgttgc cagggcaacg gggcaatacc tgtctcttca
      901 gaggagatga agttgccagg gtaactacat cctgtctttc tcaaggacca tcccagaatg
      961 tggcacccac tagccgttac catagcaact gcctctttgc cccacttaat cccatcccgt
     1021 ctgttaaaag ggccctatag ttggaggtgg gggaggtagg aagagcgatg atcacttgtg
     1081 gactaagttt gttcgcatcc ccttctccaa ccccctcagt acatcaccct gggggaacag
     1141 ggtccacttg ctcctgggcc cacacagtcc tgcagtattg tgtatataag gccagggcaa
     1201 agaggagcag gttttaaagt gaaaggcagg caggtgttgg ggaggcagtt accggggcaa
     1261 cgggaacagg gcgtttcgga ggtggttgcc atggggacct ggatgctgac gaaggctcgc
     1321 gaggctgtga gcagccacag tgccctgctc agaagcccca agctcgtcag tcaagccggt
     1381 tctccgtttg cactcaggag cacgggcagg cgagtggccc ctagttctgg gggcagctct
     1441 agagcggtac cggatccgcc accatggcat cacaggtcgt ctatggggaa tgggcatcaa
     1501 ctcacaccga gtgctacaat atgtccagaa tcgactccac cttcgtctca ctgctgcagc
     1561 tggtgtgggc cgtggtctca ggctgccaga caatcttcat gattagcagg gctcccaagg
     1621 tcccttggga gtccgtgtac ctgccctttg tcgaatctat cacttatgca ctggccagta
     1681 ctggcaacgg aaccctgcag atgagggacg gcagattctt tccttggagc cgaatggcct
     1741 cctggctgtg cacctgtcca atcatgctgg gacagatttc taacatggct ctggtgaagt
     1801 acaaaagtat cccactgaat cccattgcac aggccgcttc catcattaga gtggtcatgg
     1861 gaatcaccgc tacaatttct cccgcagagt atatgaagtg gctgttcttt ttctttgggg
     1921 ccacctgcct ggtgttcgag tacagtgtgg tcttcacaat ctttcaagtg gggctgtatg
     1981 gctttgaatc tgtcgggaca cctctggcac agaaggtggt ggtgaggatc aagatgctgc
     2041 gcctgatctt ctttattgct tggactatgt tcccaatcgt gtggctgatt tcacccaccg
     2101 gagtctgcgt gatccacgag aacgtgagcg ccattctgta cctgctggct gacggcctgt
     2161 gcaaaaatac atatggagtg atcctgtgga gcactgcctg gggagtgctg gaagggaagt
     2221 gggaccccgc atgtctgcca gggcaggaga aacccgaagc cgacgatcct tttggcctga
     2281 atcacgagaa gaatgcccct cctaacgacg aagtcaatat cagaatgttt ggaagggccg
     2341 ccaagagcag gatcaccagc gagggcgagt acatccccct ggaccagatc gacatcaacg
     2401 tggtgatggt gagcgagctg attaaggaga acatgcccat gaagctgtac atggagggca
     2461 ccgtgaacaa ccaccacttc aagtgcacat ccgagggcga aggcaagccc tacgagggca
     2521 cccagaccat gagaatcaag gtcgtcgagg gcggccctct ccccttcgcc ttcgacatcc
     2581 tggctaccag cttcatgtac ggcagcagaa ccttcatcaa gcaccctccg ggcatccccg
     2641 acttctttaa gcagtccttc cctgagggct tcacatggga gagagtcacc acatacgaag
     2701 acgggggcgt gctgaccgct acccaggaca ccagcctcca ggacggctgc ctcatctaca
     2761 acgtcaaggt tagaggggtg aacttcccag ccaacggccc tgtgatgcag aagaaaacac
     2821 tcggctggga ggcctccacc gagacgatgt accctgctga cggaggactg gaaggcgcat
     2881 gtgacatggc tctgaagctc gtgggaggag gacacctgat ctgcaacctt gagacaacat
     2941 acagatccaa gaaacccgct acgaacctca agatgcccgg cgtctacaac gtggaccaca
     3001 gactggaaag aatcaaggag gccgacgatg agacatacgt cgagcagcac gaggtggctg
     3061 tggccagata ctctactggt ggcgctggtg atggaggtaa acagtcccag cctattctga
     3121 acaccaaaga gatggccccc cagagtaaac cccctgagga gctggaaatg tcctcaatgc
     3181 cctcccctgt ggcacctctt cctgcaagaa cagagggcgt gatcgacatg aggtccatga
     3241 gttctattga ctcttttatc tcatgcgcca ccgattttcc agaggctacg cggtttaagt
     3301 aagaattcga tatcaagctt atcgataatc aacctctgga ttacaaaatt tgtgaaagat
     3361 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     3421 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     3481 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca
     3541 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt
     3601 ccggaacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg
     3661 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga
     3721 aatcatcgtc ctttccttgg ctgctcgcct atgttgccac ctggattctg cgcgggacgt
     3781 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc
     3841 cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt
     3901 gggccgcctc cccgcatcga taccgagcgc tgctcgagag atctacgggt ggcatccctg
     3961 tgacccctcc ccagtgcctc tcctggccct ggaagttgcc actccagtgc ccaccagcct
     4021 tgtcctaata aaattaagtt gcatcatttt gtctgactag gtgtccttct ataatattat
     4081 ggggtggagg ggggtggtat ggagcaaggg gcaagttggg aagacaacct gtagggcctg
     4141 cggggtctat tgggaaccaa gctggagtgc agtggcacaa tcttggctca ctgcaatctc
     4201 cgcctcctgg gttcaagcga ttctcctgcc tcagcctccc gagttgttgg gattccaggc
     4261 atgcatgacc aggctcagct aatttttgtt tttttggtag agacggggtt tcaccatatt
     4321 ggccaggctg gtctccaact cctaatctca ggtgatctac ccaccttggc ctcccaaatt
     4381 gctgggatta caggcgtgaa ccactgctcc cttccctgtc cttctgattt tgtaggtaac
     4441 cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg
     4501 cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc
     4561 cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat
     4621 tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg
     4681 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     4741 cacttgccag cgccttagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     4801 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     4861 ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat
     4921 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     4981 tcttgttcca aactggaaca acactcaact ctatctcggg ctattctttt gatttataag
     5041 ggattttgcc gatttcggtc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     5101 cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct
     5161 ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac
     5221 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     5281 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac
     5341 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     5401 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     5461 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     5521 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     5581 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     5641 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     5701 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     5761 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     5821 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     5881 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     5941 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     6001 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     6061 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     6121 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     6181 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     6241 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     6301 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     6361 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     6421 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     6481 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     6541 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     6601 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     6661 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     6721 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     6781 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     6841 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     6901 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     6961 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     7021 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     7081 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     7141 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
//
