LOCUS       sequence-369600-        6391 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses green acetylcholine indicator AchLightG in neuronal cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6391)
  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 6391)
  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_AchLightG
FEATURES             Location/Qualifiers
     source          1..6391
                     /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"
     primer_bind     1897..1918
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2024..2045)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2285..2304)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    2770..3358
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2823..2843)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3241..3252)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3376..3600
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3643..3783
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3643..3772
                     /label=AAV2 ITR
     rep_origin      3858..4313
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3945..3964)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4155..4176
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4330..4349)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4449..4471
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        4595..4699
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4700..4768,4769..5560)
                     /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(4918..4937)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5731..6319
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6220..6239
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   join(6381..6391,1..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 tacttttcca tcacgaggcc gctcacgtac cgagccaaac gaacaacaaa
     1441 gagagccggt gtgatgatcg gtctggcttg ggtcatctcc tttgtccttt gggctcctgc
     1501 catcttgttc tggcaatact ttgttggaaa gagaactgtg cctccgggag agtgcttcat
     1561 tcagttcctc agtgagccca ccattacttt tggcacagcc atcgctgctt tttatatgcc
     1621 tgtcaccatt atgactattt tatactggag gatctatagg attgctcaga aacagctgag
     1681 ctcactcatt aacgtctata tcaaggccga caagcagaag aacggcatca aggcgaactt
     1741 caagatccgc cacaacatcg aggacggcgg cgtgcagctc gcctaccact accagcagaa
     1801 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcgtgcagtc
     1861 caaactttcg aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
     1921 cgccgccggg atcactctcg gcatggacga gctgtacaag ggcggtaccg gagggagcat
     1981 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
     2041 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggt gagggcgatg ccacctacgg
     2101 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
     2161 cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca
     2221 gcacgacttc ttcaagtccg ccatgcccga aggctacatc caggagcgca ccatcttctt
     2281 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     2341 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     2401 gctggagtac aacaatcatg accaactgaa aagagaaact aaagtcgcca agaccctcag
     2461 tgcgatcttg cttgccttca tcatcacttg gaccccatac aacatcatgg ttctggtgaa
     2521 caccttttgt gacagctgca tacccaaaac cttttggaat ctgggctact ggctgtgcta
     2581 catcaacagc accgtgaacc ccgtgtgcta tgctctgtgc aacaaaacat tcagaaccac
     2641 tttcaagatg ctgctgctgt gccagtgtga caaaaaaaag aggcgcaagc agcagtacca
     2701 gcagagacag tcggtcattt ttcacaagcg cgcacccgag caggccttgt aggcggcaag
     2761 cttatcgata atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     2821 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     2881 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     2941 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     3001 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     3061 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     3121 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct
     3181 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     3241 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     3301 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgctc
     3361 tcgagatacg tacgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     3421 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     3481 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     3541 gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg
     3601 cttctgaggc ggaaagatct cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg
     3661 agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg
     3721 cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc
     3781 aggggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac
     3841 gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     3901 tacgcgcagc gtgaccgcta cacttgccag cgccttagcg cccgctcctt tcgctttctt
     3961 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     4021 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga
     4081 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     4141 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaact ctatctcggg
     4201 ctattctttt gatttataag ggattttgcc gatttcggtc tattggttaa aaaatgagct
     4261 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg
     4321 cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac
     4381 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     4441 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     4501 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     4561 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     4621 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     4681 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     4741 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     4801 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     4861 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     4921 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     4981 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     5041 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     5101 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     5161 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     5221 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     5281 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     5341 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     5401 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     5461 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     5521 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     5581 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     5641 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     5701 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     5761 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     5821 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     5881 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     5941 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     6001 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     6061 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     6121 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     6181 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     6241 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     6301 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     6361 ctggcctttt gctcacatgt cctgcaggca g
//
