LOCUS       sequence-405383-        6138 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Panneuronal expression of the MsACR1 mutant raACR fused to 
            mCerulean3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6138)
  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 6138)
  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-mCerulean3-pAAV2-hSyn
FEATURES             Location/Qualifiers
     source          1..6138
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   55..184
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        395..842
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             join(1584..1586,1587..2300)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAIHGNVYITADKQKNGIKA
                     NFGLNCNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1626..1647)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1887..1906)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2234..2255
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2325..2913
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2378..2398)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2796..2807)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2933..3409
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3132..3151)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3455..3595
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3455..3584
                     /label=AAV2 ITR
     rep_origin      3670..4125
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3757..3776)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3967..3988
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4142..4161)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4261..4283
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4321..4339)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4407..4511
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4512..4580,4581..5372)
                     /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(4730..4749)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5543..6131
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6032..6051
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgtcctgca
       61 ggcagctgcg cgctcgctcg ctcactgagg ccgcccgggc gtcgggcgac ctttggtcgc
      121 ccggcctcag tgagcgagcg agcgcgcaga gagggagtgg ccaactccat cactaggggt
      181 tcctgcggcc gcacgcgttg caaagatgga taaagtttta aacagagagg aatctttgca
      241 gctaatggac cttctaggtc ttgaaaggag tgggaattgg ctccggtgcc cgtcagtggg
      301 cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc agcaaatggt
      361 taattaatct agactgcaga gggccctgcg tatgagtgca agtgggtttt aggaccagga
      421 tgaggcgggg tgggggtgcc tacctgacga ccgaccccga cccactggac aagcacccaa
      481 cccccattcc ccaaattgcg catcccctat cagagagggg gaggggaaac aggatgcggc
      541 gaggcgcgtg cgcactgcca gcttcagcac cgcggacagt gccttcgccc ccgcctggcg
      601 gcgcgcgcca ccgccgcctc agcactgaag gcgcgctgac gtcactcgcc ggtcccccgc
      661 aaactcccct tcccggccac cttggtcgcg tccgcgccgc cgccggccca gccggaccgc
      721 accacgcgag gcgcgagata ggggggcacg ggcgcgacca tctgcgctgc ggcgccggcg
      781 actcagcgct gcctcagtct gcggtgggca gcggaggagt cgtgtcgtgc ctgagagcgc
      841 agtcgagagg atccgccacc atgtccgcct acgaggtggt gaaggacatc cggctggtca
      901 tccagaccat cctgctgctg gccttttatg tgatggccgg cttctacctg tatcagaaga
      961 agaagaagag ggccagatgg gaggtgctgt acatcgcaac cgtggcaggc ctggccaaca
     1021 caatcggcct ggccgcagga cacctggagc cactgtcttt tgccgcagag ggagtgggca
     1081 agaagacacc atggctgcgg ttcagctcct ggatgaccac atgccccgtg ctgctgatcc
     1141 acctgaacaa tctggcagga gaggaggtgc agaacgtgga gcgcgccagc atgctggtca
     1201 tcgagctgaa tctgatggtg ctgctgggcg ccacctctag cttcacagat ggcggcggcg
     1261 ccctggtgtt ctttatgctg agctggctgg tgtgctttac catcttcgtg caggcctgga
     1321 caatctttaa cgaggccttc gcctccctgc cagcaaaggc aaggcctcac ctgaatggca
     1381 tgatgttctg tttttacttc gcctgggcag gatttggcgt gctgttcgca gcaggaccag
     1441 agggatgggg acccaaggac aatatcctgg ccgagctggt gaccatcgcc atggtggtgg
     1501 tggatttcat cgccaagatc gtgtttggct tcatcggatg gcacctgagg tggcgcgtgc
     1561 tgaggggaga ggacggcaag gtggtgagca agggcgagga gctgttcacc ggggtggtgc
     1621 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
     1681 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
     1741 tgcccgtgcc ctggcccacc ctcgtgacca ccctgagctg gggcgtgcag tgcttcgccc
     1801 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     1861 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     1921 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     1981 acggcaacat cctggggcac aagctggagt acaacgccat ccacggcaac gtctatatca
     2041 ccgccgacaa gcagaagaac ggcatcaagg ccaacttcgg cctcaactgc aacatcgagg
     2101 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     2161 tgctgctgcc cgacaaccac tacctgagca cccagtccaa gctgagcaaa gaccccaacg
     2221 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
     2281 tggacgagct gtacaagtaa gaattccgta caaagcttat cgataatcaa cctctggatt
     2341 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     2401 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
     2461 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc
     2521 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca
     2581 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac
     2641 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
     2701 ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctat gttgccacct
     2761 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc
     2821 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga
     2881 cgagtcggat ctccctttgg gccgcctccc cgcatcgata ccggctcgag acgggtggca
     2941 tccctgtgac ccctccccag tgcctctcct ggccctggaa gttgccactc cagtgcccac
     3001 cagccttgtc ctaataaaat taagttgcat cattttgtct gactaggtgt ccttctataa
     3061 tattatgggg tggagggggg tggtatggag caaggggcaa gttgggaaga caacctgtag
     3121 ggcctgcggg gtctattggg aaccaagctg gagtgcagtg gcacaatctt ggctcactgc
     3181 aatctccgcc tcctgggttc aagcgattct cctgcctcag cctcccgagt tgttgggatt
     3241 ccaggcatgc atgaccaggc tcagctaatt tttgtttttt tggtagagac ggggtttcac
     3301 catattggcc aggctggtct ccaactccta atctcaggtg atctacccac cttggcctcc
     3361 caaattgctg ggattacagg cgtgaaccac tgctcccttc cctgtccttc tcgagctgat
     3421 tttgtaggta accacgtgcg gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc
     3481 actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc
     3541 ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc
     3601 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc
     3661 aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     3721 gcgtgaccgc tacacttgcc agcgccttag cgcccgctcc tttcgctttc ttcccttcct
     3781 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     3841 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac
     3901 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     3961 ttaatagtgg actcttgttc caaactggaa caacactcaa ctctatctcg ggctattctt
     4021 ttgatttata agggattttg ccgatttcgg tctattggtt aaaaaatgag ctgatttaac
     4081 aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca
     4141 gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg
     4201 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     4261 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg
     4321 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     4381 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     4441 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     4501 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     4561 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     4621 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     4681 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     4741 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     4801 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     4861 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     4921 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     4981 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     5041 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     5101 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     5161 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     5221 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     5281 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     5341 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     5401 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     5461 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     5521 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     5581 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     5641 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     5701 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     5761 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     5821 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     5881 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     5941 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     6001 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     6061 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     6121 gcctatggaa aaacgcca
//
