LOCUS       sequence-369599-        6394 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses red acetylcholine indicator AchLightR in neuronal cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6394)
  AUTHORS   Kagiampaki Z, Rohner V, Kiss C, Curreli S, Dieter A, Wilhelm M, 
            Harada M, Duss SN, Dernic J, Bhat MA, Zhou X, Ravotto L, Ziebarth T,
            Wasielewski LM, Sonmez L, Benke D, Weber B, Bohacek J, Reiner A, 
            Wiegert JS, Fellin T, Patriarchi T
  TITLE     Sensitive multicolor indicators for monitoring norepinephrine in 
            vivo.
  JOURNAL   Nat Methods. 2023 Sep;20(9):1426-1436. doi: 
            10.1038/s41592-023-01959-z.
  PUBMED    37474807
REFERENCE   2  (bases 1 to 6394)
  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"; doi: "10"; journalName: 
            "Nat Methods"; date: "2023-09"; volume: "20"; issue: "9"; pages: 
            "1426-1436"
COMMENT     Sequence Label: pAAV_hSyn1_AchLightR
FEATURES             Location/Qualifiers
     source          1..6394
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        164..611
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     intron          635..767
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     750..769
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      800..809
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     806..865
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     protein_bind    869..893
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    2773..3361
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2826..2846)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3244..3255)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3379..3603
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3646..3786
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3646..3775
                     /label=AAV2 ITR
     rep_origin      3861..4316
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3948..3967)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4158..4179
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4333..4352)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4452..4474
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4512..4530)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4598..4702
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4703..4771,4772..5563)
                     /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(4921..4940)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5734..6322
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6223..6242
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   join(6384..6394,1..119)
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcgtcgg gcgacctttg gtcgcccggc
       61 ctcagtgagc gagcgagcgc gcagagaggg agtggccaac tccatcacta ggggttcctg
      121 cggccgcacg cgtgtgtcta gactgcagag ggccctgcgt atgagtgcaa gtgggtttta
      181 ggaccaggat gaggcggggt gggggtgcct acctgacgac cgaccccgac ccactggaca
      241 agcacccaac ccccattccc caaattgcgc atcccctatc agagaggggg aggggaaaca
      301 ggatgcggcg aggcgcgtgc gcactgccag cttcagcacc gcggacagtg ccttcgcccc
      361 cgcctggcgg cgcgcgccac cgccgcctca gcactgaagg cgcgctgacg tcactcgccg
      421 gtcccccgca aactcccctt cccggccacc ttggtcgcgt ccgcgccgcc gccggcccag
      481 ccggaccgca ccacgcgagg cgcgagatag gggggcacgg gcgcgaccat ctgcgctgcg
      541 gcgccggcga ctcagcgctg cctcagtctg cggtgggcag cggaggagtc gtgtcgtgcc
      601 tgagagcgca gtcgagatta attaactcga gcaggtaagt atcaaggtta caagacaggt
      661 ttaaggagac caatagaaac tgggcttgtc gagacagaga agactcttgc gtttctgata
      721 ggcacctatt ggtcttactg acatccactt tgcctttctc tccacaggtg ctagcgttta
      781 aacggtaccg gatccgcttg ccaccatgga gacagacaca ctcctgctat gggtactgct
      841 gctctgggtt ccaggttcca ctggtgacac aagtttgtac aaaaaagttg gcaccacctt
      901 gcacaataac agtacaacct cgcctttgtt tccaaacatc agctcctcct ggatacacag
      961 cccctccgat gcagggctgc ccccgggaac cgtcactcat ttcggcagct acaatgtttc
     1021 tcgagcagct ggcaatttct cctctccaga cggtaccacc gatgaccctc tgggaggtca
     1081 taccgtctgg caagtggtct tcatcgcttt cttaacgggc atcctggcct tggtgaccat
     1141 catcggcaac atcctggtaa ttgtgtcatt taaggtcaac aagcagctga agacagtcaa
     1201 caactacttc ctcttaagcc tggcctgtgc cgatctgatt atcggggtca tttcaatgaa
     1261 tctgtttacg acctacatca tcatgaatcg atgggcctta gggaacttgg cctgtgacct
     1321 ctggcttgcc attgactacg tagccagcaa tgcctctgtt atgaatcttc tggtcatcag
     1381 ctttgacaga tactgggcga ttagttcacc cgagcgctac gagagaaaga tgactccgaa
     1441 ggcggcgttc attatgatcg gtctggcttg ggtcatctcc tttgtccttt gggctcctgc
     1501 catcttgttc tggcaatact ttgttggaaa gagaactgtg cctccgggag agtgcttcat
     1561 tcagttcctc agtgagccca ccattacttt tggcacagcc atcgctgctt tttatatgcc
     1621 tgtcaccatt atgactattt tatactggag gatctatcgg atcgcgcaga aacagctctc
     1681 cagcttccac gtgagcgaga ggatgtatcc tgaggacggt gcgctgaaat ccgaaataaa
     1741 gaaaggcctg cgcctcaaag acgggggcca ctacgccgct gaggttaaga caacttacaa
     1801 ggctaaaaag cctgtccagc tcccgggcgc ctacatagtc gacattaaac tggacatagt
     1861 cagccataac gaggactaca ccatagtcga acagtgtgag cgggcggaag gtcgccacag
     1921 tacaggagga atggacgaat tgtataaagg gggcaccggt ggcagcttgg ttagcaaggg
     1981 agaggaagat aacatggcta ttataaagga gttcatgaga tttaaagttc acatggaggg
     2041 atctgtaaac gggcatgagt ttgagattga gggagaagga gaagggcgac cgtacgaggc
     2101 gttccaaacc gcgaaattga aggtcaccaa gggaggaccc cttccgtttg cttgggacat
     2161 tcttagccca caattcatgt atggcagcaa ggcgtacata aagcacccgg cagacatacc
     2221 cgactacttc aaacttagct ttccagaggg tttccgatgg gaaagagtga tgaactttga
     2281 ggatggtggc atcatacacg tcaatcaaga tagcagcctt caggacggag tatttatata
     2341 caaagtcaaa ctccggggta caaactttcc cccggacggg ccagtcatgc agaagaagac
     2401 tatgggttgg gaggcgcgga aagacgactt gaagagggag actaaagtcc tcaagaccct
     2461 cagtgcgatc ttgcttgcct tcatcatcac ttggacccca tacaacatca tggttctggt
     2521 gaacaccttt tgtgacagct gcatacccaa aaccttttgg aatctgggct actggctgtg
     2581 ctacatcaac agcaccgtga accccgtgtg ctatgctctg tgcaacaaaa cattcagaac
     2641 cactttcaag atgctgctgc tgtgccagtg tgacaaaaaa aagaggcgca agcagcagta
     2701 ccagcagaga cagtcggtca tttttcacaa gcgcgcaccc gagcaggcct tgtaggcggc
     2761 aagcttatcg ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     2821 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     2881 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
     2941 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
     3001 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
     3061 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     3121 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt
     3181 ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
     3241 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
     3301 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
     3361 ctctcgagat acgtacgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     3421 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     3481 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     3541 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta
     3601 tggcttctga ggcggaaaga tctcacgtgc ggaccgagcg gccgcaggaa cccctagtga
     3661 tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg
     3721 tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc
     3781 tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     3841 tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     3901 ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt
     3961 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     4021 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg
     4081 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     4141 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca actctatctc
     4201 gggctattct tttgatttat aagggatttt gccgatttcg gtctattggt taaaaaatga
     4261 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg
     4321 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
     4381 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     4441 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     4501 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     4561 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     4621 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     4681 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     4741 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     4801 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     4861 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     4921 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     4981 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     5041 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     5101 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     5161 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     5221 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     5281 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     5341 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     5401 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     5461 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     5521 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     5581 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     5641 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     5701 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     5761 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     5821 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     5881 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     5941 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     6001 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     6061 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     6121 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     6181 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     6241 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     6301 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     6361 ttgctggcct tttgctcaca tgtcctgcag gcag
//
