LOCUS       sequence-446874-        5589 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0780m is an enhancer sequence, designed to drive AAV-mediated 
            transgene expression in striatal D1 MSNs.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5589)
  AUTHORS   Hunker AC, Wirthlin ME, Gill G, Johansen NJ, Hooper M, Omstead V, 
            Vargas S, Lerma MN, Taskin N, Weed N, Laird WD, Bishaw YM, Bendrick 
            JL, Gore BB, Ben-Simon Y, Opitz-Araya X, Martinez RA, Way SW, 
            Thyagarajan B, Otto S, Sanchez REA, Alexander JR, Amaya A, Amster A,
            Arbuckle J, Ayala A, Baker PM, Barcelli T, Barta S, Bertagnolli D, 
            Bielstein C, Bishwakarma P, Bowlus J, Boyer G, Brouner K, Casian B, 
            Casper T, Chakka AB, Chakrabarty R, Chong P, Clark M, Colbert K, 
            Daniel S, Dawe T, Departee M, DiValentin P, Donadio NP, Dotson NI, 
            Dwivedi D, Egdorf T, Fliss T, Gary A, Goldy J, Grasso C, Groce EL, 
            Gudsnuk K, Han W, Haradon Z, Hastings S, Helback O, Ho WV, Huang C, 
            Johnson T, Jones DL, Juneau Z, Kenney J, Leibly M, Li S, Liang E, 
            Loeffler H, Lusk NA, Madigan Z, Malloy J, Malone J, McCue R, Melchor
            J, Mich JK, Moosman S, Morin E, Naidoo R, Newman D, Ngo K, Nguyen K,
            Oster AL, Ouellette B, Oyama AA, Pena N, Pham T, Phillips E, Pom C, 
            Potekhina L, Ransford S, Ray PL, Reding M, Rette DF, Reynoldson C, 
            Rimorin C, Sigler AR, Rocha DB, Ronellenfitch K, Ruiz A, Sawyer L, 
            Sevigny JP, Shapovalova NV, Shepard N, Shulga L, Soliman S, Staats 
            B, Taormina MJ, Tieu M, Wang Y, Wilkes J, Wood T, Zhou T, Williford 
            A, Dee N, Mollenkopf T, Ng L, Esposito L, Kalmbach BE, Yao S, Ariza 
            J, Collman F, Mufti S, Smith K, Waters J, Ersing I, Patrick M, Zeng 
            H, Lein ES, Kojima Y, Horwitz G, Owen SF, Levi BP, Daigle TL, Tasic 
            B, Bakken TE, Ting JT
  TITLE     Enhancer AAV toolbox for accessing and perturbing striatal cell 
            types and circuits.
  JOURNAL   Neuron. 2025 May 21;113(10):1507-1524.e17. doi: 
            10.1016/j.neuron.2025.04.035.
  PUBMED    40403704
REFERENCE   2  (bases 1 to 5589)
  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: 
            "10.1016/j.neuron.2025.04"; journalName: "Neuron"; date: 
            "2025-05-21- 21"; volume: "113"; issue: "10"; pages: "1507-1524.e17"
COMMENT     Sequence Label: AiP15579 -
            pAAV-AiE0780m-minBG-CoChR-EGFP-WPRE...
FEATURES             Location/Qualifiers
     source          1..5589
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     RBS             364..372
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             750..1613
                     /codon_start=1
                     /product="high-photocurrent channelrhodopsin from 
                     Chloromonas oogama (Shemesh et al., 2017)"
                     /label=CoChR
                     /translation="MLGNGSAIVPIDQCFCLAWTDSLGSDTEQLVANILQWFAFGFSIL
                     ILMFYAYQTWRATCGWEEVYVCCVELTKVIIEFFHEFDDPSMLYLANGHRVQWLRYAEW
                     LLTCPVILIHLSNLTGLKDDYSKRTMRLLVSDVGTIVWGATSAMSTGYVKVIFFVLGCI
                     YGANTFFHAAKVYIESYHVVPKGRPRTVVRIMAWLFFLSWGMFPVLFVVGPEGFDAISV
                     YGSTIGHTIIDLMSKNCWGLLGHYLRVLIHQHIIIYGDIRKKTKINVAGEEMEVETMVD
                     QEDEETV"
     CDS             join(1632..1634,1635..2348)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(1632..1634,1635..2345)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1674..1695)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1935..1954)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2282..2303
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2416..2436)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2623..2734
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2852..2992
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2852..2981
                     /label=AAV2 ITR
     rep_origin      3067..3522
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3154..3173)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3364..3385
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3539..3558)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3658..3680
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3718..3736)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3804..3908
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3909..3977,3978..4769)
                     /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(4127..4146)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4940..5528
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5429..5448
                     /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 gcgttgtttc acaaactaca ggttcactgg agcacagcca
      181 tgccaacatg gcatctaagg ctggtatgac ctcgagaccc ctccagcact ccttcgttca
      241 gccatagaac aaggagacac agataaagtc tggagccggg cccaggtacc tgaggctcag
      301 agatgagccc gcagcgaagc tgacaactga ctcattcctc cgacactctc ccaccctccc
      361 agctaaggag gtgtctgatg tgaacatgtt ctgcctcttt gttgcctgct gttgggggcg
      421 agggctttgg gtggattttg aagagcaatt atgcttatta ctggttagga tttctctgtt
      481 tgctgagtgc attaactttt agcgcttccc atcactcatt cttcttcaca ccctcttgtc
      541 agttacaaat gtgcaggtgg agagccacga aaaaaccctg cagcaacggc aacagaaatc
      601 acagcaacaa caacaaatga acagaaagga tagaaagcag atactcaaag atggagaaaa
      661 agatactccc gagaggctga gctcgggctg ggcataaaag tcagggcaga gccatctatt
      721 gcttacattt gcttctggga tccgccacca tgctgggaaa cggcagcgcc attgtgccta
      781 tcgaccagtg cttttgcctg gcttggaccg acagcctggg aagcgataca gagcagctgg
      841 tggccaacat cctccagtgg ttcgccttcg gcttcagcat cctgatcctg atgttctacg
      901 cctaccagac ttggagagcc acttgcggtt gggaggaggt ctacgtctgt tgcgtcgagc
      961 tgaccaaggt catcatcgag ttcttccacg agttcgacga ccccagcatg ctgtacctgg
     1021 ctaacggaca ccgagtccag tggctgagat acgcagagtg gctgctgact tgtcccgtca
     1081 tcctgatcca cctgagcaac ctgaccggcc tgaaggacga ctacagcaag cggaccatga
     1141 ggctgctggt gtcagacgtg ggaaccatcg tgtggggagc tacaagcgcc atgagcacag
     1201 gctacgtcaa ggtcatcttc ttcgtgctgg gttgcatcta cggcgccaac accttcttcc
     1261 acgccgccaa ggtgtatatc gagagctacc acgtggtgcc aaagggcaga cctagaaccg
     1321 tcgtgcggat catggcttgg ctgttcttcc tgtcttgggg catgttcccc gtgctgttcg
     1381 tcgtgggacc agaaggattc gacgccatca gcgtgtacgg ctctaccatt ggccacacca
     1441 tcatcgacct catgagcaag aattgttggg gcctgctggg acactatctg agagtgctga
     1501 tccaccagca catcatcatc tacggcgaca tccggaagaa gaccaagatc aacgtggccg
     1561 gcgaggagat ggaagtggag accatggtgg accaggagga cgaggagaca gtggcggcac
     1621 cggtagtagc agtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
     1681 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
     1741 ccacctacgg caagctgacc ctgaagttca tttgcaccac cggcaagctg cccgtgccct
     1801 ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc
     1861 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
     1921 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
     1981 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
     2041 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc
     2101 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
     2161 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     2221 acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc
     2281 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
     2341 acaagtaaga attcgatatc ataatcaacc tctggattac aaaatttgtg aaagattgac
     2401 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     2461 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     2521 agttcttgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     2581 gctgttgggc actgacaatt ccgtggctcg agagatcttc gactgtgcct tctagttgcc
     2641 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     2701 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     2761 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     2821 atgagatctc acgtgcggac cgagcggccg caggaacccc tagtgatgga gttggccact
     2881 ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg
     2941 ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg
     3001 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac
     3061 catagtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     3121 tgaccgctac acttgccagc gccttagcgc ccgctccttt cgctttcttc ccttcctttc
     3181 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     3241 gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta
     3301 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     3361 atagtggact cttgttccaa actggaacaa cactcaactc tatctcgggc tattcttttg
     3421 atttataagg gattttgccg atttcggtct attggttaaa aaatgagctg atttaacaaa
     3481 aatttaacgc gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta
     3541 caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg
     3601 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     3661 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc
     3721 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag
     3781 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     3841 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     3901 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     3961 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     4021 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     4081 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     4141 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     4201 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     4261 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     4321 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     4381 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     4441 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     4501 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     4561 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     4621 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     4681 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     4741 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     4801 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     4861 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     4921 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     4981 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     5041 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     5101 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     5161 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     5221 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     5281 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     5341 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     5401 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     5461 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     5521 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     5581 ctcacatgt
//
