LOCUS       sequence-459475-        5423 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0445h_3xC2 is an enhancer sequence, designed to drive 
            AAV-mediated transgene expression in striatal indirect pathway MSNs 
            (D2-MSNs).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5423)
  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 5423)
  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: AiP14133:
            pAAV-AiE0445h_3xC2-minBG-jGCaMP8m-WP...
FEATURES             Location/Qualifiers
     source          1..5423
                     /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             901..918
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     1200..1221
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1327..1348)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1588..1607)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     complement(2250..2270)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2457..2568
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2686..2826
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2686..2815
                     /label=AAV2 ITR
     rep_origin      2901..3356
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2988..3007)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3198..3219
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3373..3392)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3492..3514
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3552..3570)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3638..3742
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3743..3811,3812..4603)
                     /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(3961..3980)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4774..5362
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5263..5282
                     /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 gcgtgtggag gactttgagc ttaaccctat taaatttaat
      181 ctaatatatt cagcagcctg tcacaatctt gttaaatctt tattccgtca cccaccttat
      241 tagggggccc tcccacttta gtcatcagca gattagtaag catgccattt agatgtcatc
      301 taagttaatg aacctgactg agccagagtc acatggcaga gtcacagaaa ctccttctga
      361 gctggcgtgg aggactttga gcttaaccct attaaattta atctaatata ttcagcagcc
      421 tgtcacaatc ttgttaaatc tttattccgt cacccacctt attagggggc cctcccactt
      481 tagtcatcag cagattagta agcatgccat ttagatgtca tctaagttaa tgaacctgac
      541 tgagccagag tcacatggca gagtcacaga aactccttct gagctggcgt ggaggacttt
      601 gagcttaacc ctattaaatt taatctaata tattcagcag cctgtcacaa tcttgttaaa
      661 tctttattcc gtcacccacc ttattagggg gccctcccac tttagtcatc agcagattag
      721 taagcatgcc atttagatgt catctaagtt aatgaacctg actgagccag agtcacatgg
      781 cagagtcaca gaaactcctt ctgagctggc gagctcgggc tgggcataaa agtcagggca
      841 gagccatcta ttgcttacat ttgcttctgg gatccagatc tttcgaagct agccaccatg
      901 catcatcacc atcatcacac gcgtcgcaag aagaccttca aggaggtggc caccgccgtg
      961 aagatcatcg gccgcctgat gggcctgaag atcaacgtct atatcaaggc cgacaagcag
     1021 aagaacggca tcaaggcgaa cttccacatc cgccacaaca tcgaggacgg cggcgtgcag
     1081 ctcgcctacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     1141 aaccactacc tgagcgtgga gtccaaactt tcgaaagacc ccaacgagaa gcgcgatcac
     1201 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
     1261 aagggcggta ccggagggag catggtgagc aagggcgagg agctgttcac cggggtggtg
     1321 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     1381 ggtgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     1441 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc
     1501 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     1561 atccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     1621 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     1681 gacggcaaca tcctggggca caagctggag tacaacctgc cggaccaact gactgaagag
     1741 cagatcgcag aatacaaaga ggctttctcc ctatttgaca aggacgggga tgggacaata
     1801 acaaccaagg agctggggac ggtgatgcgg tctctggggc agaaccccac agaagcagag
     1861 ctgcaggaca tgatcaatga agtagatgcc gacggtgacg gcacaatcga cttccctgag
     1921 ttcctgacaa tgatggcaag aaaaatgaaa tacagggaca cggaagaaga aattagagaa
     1981 gcgttcggtg tgtttgataa ggatggcaat ggctacatca gtgcagcaga gcttcgccac
     2041 gtgatgacaa accttggaga gaagttaaca gatgaagagg ttgatgaaat gatcagggaa
     2101 gcagacatcg atggggatgg tcaggtaaac tacgaagagt ttgtacaaat gatgacagcg
     2161 aagtaaggcg cgccgcggcc gcgaattcga tatcataatc aacctctgga ttacaaaatt
     2221 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     2281 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     2341 tataaatcct ggttagttct tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     2401 tggacagggg ctcggctgtt gggcactgac aattccgtgg ctcgagagat cttcgactgt
     2461 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     2521 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     2581 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     2641 agacaatagc aggcatgaga tctcacgtgc ggaccgagcg gccgcaggaa cccctagtga
     2701 tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg
     2761 tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc
     2821 tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     2881 tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     2941 ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt
     3001 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     3061 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg
     3121 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     3181 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca actctatctc
     3241 gggctattct tttgatttat aagggatttt gccgatttcg gtctattggt taaaaaatga
     3301 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg
     3361 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
     3421 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     3481 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     3541 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     3601 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     3661 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     3721 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     3781 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     3841 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     3901 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     3961 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     4021 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     4081 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     4141 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     4201 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     4261 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     4321 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     4381 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     4441 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     4501 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     4561 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     4621 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     4681 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     4741 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     4801 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     4861 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     4921 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     4981 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     5041 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     5101 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     5161 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     5221 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     5281 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     5341 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     5401 ttgctggcct tttgctcaca tgt
//
