LOCUS       sequence-405388-        5571 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Soma-targeted neuronal expression of the MsACR1 mutant raACR and 
            mCerulean3 separated by a P2A site..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5571)
  AUTHORS   Oppermann J, Rozenberg A, Fabrin T, Gonzalez-Cabrera C, Parker R, 
            Beja O, Prigge M, Hegemann P
  TITLE     Robust optogenetic inhibition with red-light-sensitive 
            anion-conducting channelrhodopsins.
  JOURNAL   eLife. 2024 Oct 14;12:RP90100. doi: 10.7554/eLife.90100.
  PUBMED    39401075
REFERENCE   2  (bases 1 to 5571)
  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"; journalName: "eLife."; 
            date: "2024-10-14"; volume: "12:RP90100. doi"; pages: " 
            10.7554/eLife.90100"
COMMENT     Sequence Label: raACR-TS-Kv-P2A-mCerulean3-pAAV-CaMKII
FEATURES             Location/Qualifiers
     source          1..5571
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   20..149
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     CDS             1561..1617
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             join(1630..1632,1633..1635,1636..2349)
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1675..1696)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1936..1955)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2283..2304
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2365..2768
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     polyA_signal    2775..2823
                     /note="synthetic polyadenylation signal"
     repeat_region   2853..2993
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2853..2982
                     /label=AAV2 ITR
     rep_origin      3068..3523
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3155..3174)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3365..3386
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3540..3559)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3659..3681
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3719..3737)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3805..3909
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3910..3978,3979..4770)
                     /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(4128..4147)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4941..5529
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5430..5449
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tggccttttg ctcacatgtc ctgcaggcag ctgcgcgctc gctcgctcac tgaggccgcc
       61 cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg
      121 agtggccaac tccatcacta ggggttcctg cggccgcacg cgtcttgtgg actaagtttg
      181 ttcgcatccc cttctccaac cccctcagta catcaccctg ggggaacagg gtccacttgc
      241 tcctgggccc acacagtcct gcagtattgt gtatataagg ccagggcaaa gaggagcagg
      301 ttttaaagtg aaaggcaggc aggtgttggg gaggcagtta ccggggcaac gggaacaggg
      361 cgtttcggag gtggttgcca tggggacctg gatgctgacg aaggctcgcg aggctgtgag
      421 cagccacagt gccctgctca gaagccccaa gctcgtcagt caagccggtt ctccgtttgc
      481 actcaggagc acgggcaggc gagtggcccc tagttctggg ggcagctcta gagcggtacc
      541 ggatccgcca ccatgtccgc ctacgaggtg gtgaaggaca tccggctggt catccagacc
      601 atcctgctgc tggcctttta tgtgatggcc ggcttctacc tgtatcagaa gaagaagaag
      661 agggccagat gggaggtgct gtacatcgca accgtggcag gcctggccaa cacaatcggc
      721 ctggccgcag gacacctgga gccactgtct tttgccgcag agggagtggg caagaagaca
      781 ccatggctgc ggttcagctc ctggatgacc acatgccccg tgctgctgat ccacctgaac
      841 aatctggcag gagaggaggt gcagaacgtg gagcgcgcca gcatgctggt catcgagctg
      901 aatctgatgg tgctgctggg cgccacctct agcttcacag atggcggcgg cgccctggtg
      961 ttctttatgc tgagctggct ggtgtgcttt accatcttcg tgcaggcctg gacaatcttt
     1021 aacgaggcct tcgcctccct gccagcaaag gcaaggcctc acctgaatgg catgatgttc
     1081 tgtttttact tcgcctgggc aggatttggc gtgctgttcg cagcaggacc agagggatgg
     1141 ggacccaagg acaatatcct ggccgagctg gtgaccatcg ccatggtggt ggtggatttc
     1201 atcgccaaga tcgtgtttgg cttcatcgga tggcacctga ggtggcgcgt gctgagggga
     1261 gaggacggca aggtgggacc gaaaccggta aagagcagga tcaccagcga gggcgagtac
     1321 atccccctgg accagatcga catcaacgtg gtgcagtccc agcctattct gaacaccaaa
     1381 gagatggccc cccagagtaa accccctgag gagctggaaa tgtcctcaat gccctcccct
     1441 gtggcacctc ttcctgcaag aacagagggc gtgatcgaca tgaggtccat gagttctatt
     1501 gactctttta tctcatgcgc caccgatttt ccagaggcta cgcggtttaa gggctccgga
     1561 gccacgaact tctctctgtt aaagcaagca ggagacgtgg aagaaaaccc cggtcctgac
     1621 atgcctagga tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     1681 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
     1741 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
     1801 tggcccaccc tcgtgaccac cctgacctgg ggcgtgcagt gcttcagccg ctaccccgac
     1861 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     1921 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     1981 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     2041 ctggggcaca agctggagta caactacatc agccacaacg tctatatcac cgccgacaag
     2101 cagaagaacg gcatcaaggc caacttcaag atccgccaca acatcgagga cggcagcgtg
     2161 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     2221 gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat
     2281 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     2341 tacaagtaat aggaattcaa gcttaatcaa cctctggatt acaaaatttg tgaaagattg
     2401 actgatattc ttaactatgt tgctcctttt acgctgtgtg gatatgctgc tttaatgcct
     2461 ctgtatcatg ctattgcttc ccgtatggct ttcgttttct cctccttgta taaatcctgg
     2521 ttgctgtctc tttatgagga gttgtggccc gttgtccgtc aacgtggcgt ggtgtgctct
     2581 gtgtttgctg acgcaacccc cactggctgg ggcattgcca ccacctgtca actcctttct
     2641 gggactttcg ctttccccct cccgatcgcc acggcagaac tcatcgccgc ctgccttgcc
     2701 cgctgctgga caggggctag gttgctgggc actgataatt ccgtggtgtt gtcggggaag
     2761 ctgacgtcag cgctaataaa atcgctatcc atcgaagatg gatgtgtgtt ggttttttgt
     2821 gtgggtaacc cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg agttggccac
     2881 tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc
     2941 gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct
     3001 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa
     3061 ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     3121 gtgaccgcta cacttgccag cgccttagcg cccgctcctt tcgctttctt cccttccttt
     3181 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     3241 cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt
     3301 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     3361 aatagtggac tcttgttcca aactggaaca acactcaact ctatctcggg ctattctttt
     3421 gatttataag ggattttgcc gatttcggtc tattggttaa aaaatgagct gatttaacaa
     3481 aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt
     3541 acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac
     3601 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     3661 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc
     3721 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     3781 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     3841 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     3901 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     3961 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     4021 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     4081 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     4141 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     4201 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     4261 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     4321 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     4381 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     4441 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     4501 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     4561 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     4621 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     4681 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     4741 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     4801 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     4861 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     4921 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     4981 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     5041 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     5101 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     5161 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     5221 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     5281 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     5341 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     5401 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     5461 gggttttgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     5521 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg c
//
