LOCUS       sequence-447004-        5652 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0441h_3xC2 is an enhancer sequence, designed to drive 
            AAV-mediated transgene expression in striatal MSNs.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5652)
  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 5652)
  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: AiP15457 -
            pAAV-AiE0441h_3xC2-minBG-CoChR-EGFP...
FEATURES             Location/Qualifiers
     source          1..5652
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     CDS             813..1676
                     /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(1695..1697,1698..2411)
                     /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(1695..1697,1698..2408)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1737..1758)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1998..2017)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2345..2366
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2479..2499)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2686..2797
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2915..3055
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2915..3044
                     /label=AAV2 ITR
     rep_origin      3130..3585
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3217..3236)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3427..3448
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3602..3621)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3721..3743
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3781..3799)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3867..3971
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3972..4040,4041..4832)
                     /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(4190..4209)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5003..5591
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5492..5511
                     /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 gcgtcccaca cagcccccac cacctggccg cccacctgaa
      181 aaacttttcc ccaagttctt gagaagtcgc gtaggatcca ggagagaaaa atcaaagtaa
      241 taaaacctca tttaattcct aacggagggc agagctgcga gctgcaatta tcttaatgct
      301 gagctatttt tactctctct ccatctctgt cttctgcctt tctcccacac agcccccacc
      361 acctggccgc ccacctgaaa aacttttccc caagttcttg agaagtcgcg taggatccag
      421 gagagaaaaa tcaaagtaat aaaacctcat ttaattccta acggagggca gagctgcgag
      481 ctgcaattat cttaatgctg agctattttt actctctctc catctctgtc ttctgccttt
      541 ctcccacaca gcccccacca cctggccgcc cacctgaaaa acttttcccc aagttcttga
      601 gaagtcgcgt aggatccagg agagaaaaat caaagtaata aaacctcatt taattcctaa
      661 cggagggcag agctgcgagc tgcaattatc ttaatgctga gctattttta ctctctctcc
      721 atctctgtct tctgcctttc tgagctcggg ctgggcataa aagtcagggc agagccatct
      781 attgcttaca tttgcttctg ggatccgcca ccatgctggg aaacggcagc gccattgtgc
      841 ctatcgacca gtgcttttgc ctggcttgga ccgacagcct gggaagcgat acagagcagc
      901 tggtggccaa catcctccag tggttcgcct tcggcttcag catcctgatc ctgatgttct
      961 acgcctacca gacttggaga gccacttgcg gttgggagga ggtctacgtc tgttgcgtcg
     1021 agctgaccaa ggtcatcatc gagttcttcc acgagttcga cgaccccagc atgctgtacc
     1081 tggctaacgg acaccgagtc cagtggctga gatacgcaga gtggctgctg acttgtcccg
     1141 tcatcctgat ccacctgagc aacctgaccg gcctgaagga cgactacagc aagcggacca
     1201 tgaggctgct ggtgtcagac gtgggaacca tcgtgtgggg agctacaagc gccatgagca
     1261 caggctacgt caaggtcatc ttcttcgtgc tgggttgcat ctacggcgcc aacaccttct
     1321 tccacgccgc caaggtgtat atcgagagct accacgtggt gccaaagggc agacctagaa
     1381 ccgtcgtgcg gatcatggct tggctgttct tcctgtcttg gggcatgttc cccgtgctgt
     1441 tcgtcgtggg accagaagga ttcgacgcca tcagcgtgta cggctctacc attggccaca
     1501 ccatcatcga cctcatgagc aagaattgtt ggggcctgct gggacactat ctgagagtgc
     1561 tgatccacca gcacatcatc atctacggcg acatccggaa gaagaccaag atcaacgtgg
     1621 ccggcgagga gatggaagtg gagaccatgg tggaccagga ggacgaggag acagtggcgg
     1681 caccggtagt agcagtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
     1741 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
     1801 atgccaccta cggcaagctg accctgaagt tcatttgcac caccggcaag ctgcccgtgc
     1861 cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg
     1921 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
     1981 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
     2041 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
     2101 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca
     2161 agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg
     2221 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     2281 ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg
     2341 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
     2401 tgtacaagta agaattcgat atcataatca acctctggat tacaaaattt gtgaaagatt
     2461 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     2521 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     2581 gttagttctt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     2641 tcggctgttg ggcactgaca attccgtggc tcgagagatc ttcgactgtg ccttctagtt
     2701 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     2761 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     2821 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     2881 ggcatgagat ctcacgtgcg gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc
     2941 actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc
     3001 ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc
     3061 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc
     3121 aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     3181 gcgtgaccgc tacacttgcc agcgccttag cgcccgctcc tttcgctttc ttcccttcct
     3241 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     3301 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac
     3361 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     3421 ttaatagtgg actcttgttc caaactggaa caacactcaa ctctatctcg ggctattctt
     3481 ttgatttata agggattttg ccgatttcgg tctattggtt aaaaaatgag ctgatttaac
     3541 aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca
     3601 gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg
     3661 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     3721 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg
     3781 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     3841 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     3901 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     3961 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     4021 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     4081 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     4141 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     4201 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     4261 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     4321 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     4381 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     4441 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     4501 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     4561 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     4621 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     4681 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     4741 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     4801 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     4861 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     4921 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     4981 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     5041 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     5101 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     5161 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     5221 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     5281 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     5341 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     5401 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     5461 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     5521 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     5581 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     5641 ttgctcacat gt
//
