LOCUS       sequence-446786-        6468 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0784m is an enhancer sequence, designed to drive Cre dependent 
            AAV-mediated transgene expression in striatal MSNs.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6468)
  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 6468)
  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: AiP15356 -
            pAAV-AiE0784m-minBG-DIO-CoChR-EGFP-...
FEATURES             Location/Qualifiers
     source          1..6468
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     protein_bind    complement(718..751)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(802..835)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(join(844..1557,1558..1560))
                     /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             complement(join(847..1557,1558..1560))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(889..910)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1238..1257
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1497..1518
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(1579..2442)
                     /codon_start=1
                     /product="high-photocurrent channelrhodopsin from 
                     Chloromonas oogama (Shemesh et al., 2017)"
                     /label=CoChR
                     /translation="MLGNGSAIVPIDQCFCLAWTDSLGSDTEQLVANILQWFAFGFSIL
                     ILMFYAYQTWRATCGWEEVYVCCVELTKVIIEFFHEFDDPSMLYLANGHRVQWLRYAEW
                     LLTCPVILIHLSNLTGLKDDYSKRTMRLLVSDVGTIVWGATSAMSTGYVKVIFFVLGCI
                     YGANTFFHAAKVYIESYHVVPKGRPRTVVRIMAWLFFLSWGMFPVLFVVGPEGFDAISV
                     YGSTIGHTIIDLMSKNCWGLLGHYLRVLIHQHIIIYGDIRKKTKINVAGEEMEVETMVD
                     QEDEETV"
     protein_bind    2453..2486
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    2537..2570
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2595..3183
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2648..2668)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3066..3077)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3215..3691
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3414..3433)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3731..3871
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3731..3860
                     /label=AAV2 ITR
     rep_origin      3946..4401
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4033..4052)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4243..4264
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4418..4437)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4537..4559
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4597..4615)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4683..4787
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4788..4856,4857..5648)
                     /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(5006..5025)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5819..6407
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6308..6327
                     /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 gcgtaggaac tggacacaca aagcctttct cgttttttct
      181 tcctccctgt gtttcggtcc aggggtccta agtgtgtagc tcccggggag atgtgggact
      241 gttctcaccc agaaagagga actagaacca caagagccta accccccacg gtgggaagac
      301 actctgtggc atcaccggga ggggaggcca ggtgtgggca actgcagcta tttctaatcc
      361 accagaggaa agagagcttt gactgcttga aaagcaggct gattgtcagg ggcggaaccc
      421 agctcagagc tgtcagggaa gctcctggca gggggtggag acagggggct tggactgaca
      481 ggcctgtgat tttgaaagaa atgtctttaa gagagtgact taaagtagga gagaaatatt
      541 attattaggt aatgcctgtc aggttatatt tttttttgat gtgttcttgg tatttcataa
      601 tcattttgcc tttagatggt aagcccttcc tggtgagctc gggctgggca taaaagtcag
      661 ggcagagcca tctattgctt acatttgctt ctgggatcct ctagagtcga ctccggaata
      721 acttcgtata ggatacttta tacgaagtta tgcagaatgg tagctggatt gtagctgcta
      781 ttagcaatat gaaacctctt aataacttcg tatagcatac attatacgaa gttatggcgc
      841 gccttacttg tacagctcgt ccatgccgag agtgatcccg gcggcggtca cgaactccag
      901 caggaccatg tgatcgcgct tctcgttggg gtctttgctc agggcggact gggtgctcag
      961 gtagtggttg tcgggcagca gcacggggcc gtcgccgatg ggggtgttct gctggtagtg
     1021 gtcggcgagc tgcacgctgc cgtcctcgat gttgtggcgg atcttgaagt tcaccttgat
     1081 gccgttcttc tgcttgtcgg ccatgatata gacgttgtgg ctgttgtagt tgtactccag
     1141 cttgtgcccc aggatgttgc cgtcctcctt gaagtcgatg cccttcagct cgatgcggtt
     1201 caccagggtg tcgccctcga acttcacctc ggcgcgggtc ttgtagttgc cgtcgtcctt
     1261 gaagaagatg gtgcgctcct ggacgtagcc ttcgggcatg gcggacttga agaagtcgtg
     1321 ctgcttcatg tggtcggggt agcggctgaa gcactgcacg ccgtaggtca gggtggtcac
     1381 gagggtgggc cagggcacgg gcagcttgcc ggtggtgcaa atgaacttca gggtcagctt
     1441 gccgtaggtg gcatcgccct cgccctcgcc ggacacgctg aacttgtggc cgtttacgtc
     1501 gccgtccagc tcgaccagga tgggcaccac cccggtgaac agctcctcgc ccttgctcac
     1561 tgctactacc ggtgccgcca ctgtctcctc gtcctcctgg tccaccatgg tctccacttc
     1621 catctcctcg ccggccacgt tgatcttggt cttcttccgg atgtcgccgt agatgatgat
     1681 gtgctggtgg atcagcactc tcagatagtg tcccagcagg ccccaacaat tcttgctcat
     1741 gaggtcgatg atggtgtggc caatggtaga gccgtacacg ctgatggcgt cgaatccttc
     1801 tggtcccacg acgaacagca cggggaacat gccccaagac aggaagaaca gccaagccat
     1861 gatccgcacg acggttctag gtctgccctt tggcaccacg tggtagctct cgatatacac
     1921 cttggcggcg tggaagaagg tgttggcgcc gtagatgcaa cccagcacga agaagatgac
     1981 cttgacgtag cctgtgctca tggcgcttgt agctccccac acgatggttc ccacgtctga
     2041 caccagcagc ctcatggtcc gcttgctgta gtcgtccttc aggccggtca ggttgctcag
     2101 gtggatcagg atgacgggac aagtcagcag ccactctgcg tatctcagcc actggactcg
     2161 gtgtccgtta gccaggtaca gcatgctggg gtcgtcgaac tcgtggaaga actcgatgat
     2221 gaccttggtc agctcgacgc aacagacgta gacctcctcc caaccgcaag tggctctcca
     2281 agtctggtag gcgtagaaca tcaggatcag gatgctgaag ccgaaggcga accactggag
     2341 gatgttggcc accagctgct ctgtatcgct tcccaggctg tcggtccaag ccaggcaaaa
     2401 gcactggtcg ataggcacaa tggcgctgcc gtttcccagc atggtggcta gcataacttc
     2461 gtataaagta tcctatacga agttatttgc cttaacccag aaattatcac tgttattctt
     2521 tagaatggtg caaagaataa cttcgtataa tgtatgctat acgaagttat gaattcgata
     2581 tcaagcttat cgataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc
     2641 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
     2701 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc
     2761 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg
     2821 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg
     2881 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga
     2941 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct
     3001 ttccttggct gctcgcctat gttgccacct ggattctgcg cgggacgtcc ttctgctacg
     3061 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc
     3121 ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc
     3181 cgcatcgata ccgagcgctg ctcgagagat ctacgggtgg catccctgtg acccctcccc
     3241 agtgcctctc ctggccctgg aagttgccac tccagtgccc accagccttg tcctaataaa
     3301 attaagttgc atcattttgt ctgactaggt gtccttctat aatattatgg ggtggagggg
     3361 ggtggtatgg agcaaggggc aagttgggaa gacaacctgt agggcctgcg gggtctattg
     3421 ggaaccaagc tggagtgcag tggcacaatc ttggctcact gcaatctccg cctcctgggt
     3481 tcaagcgatt ctcctgcctc agcctcccga gttgttggga ttccaggcat gcatgaccag
     3541 gctcagctaa tttttgtttt tttggtagag acggggtttc accatattgg ccaggctggt
     3601 ctccaactcc taatctcagg tgatctaccc accttggcct cccaaattgc tgggattaca
     3661 ggcgtgaacc actgctccct tccctgtcct tctgattttg taggtaacca cgtgcggacc
     3721 gagcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg
     3781 ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca
     3841 gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg
     3901 catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg
     3961 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     4021 ccttagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     4081 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     4141 tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga
     4201 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     4261 ctggaacaac actcaactct atctcgggct attcttttga tttataaggg attttgccga
     4321 tttcggtcta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     4381 aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat
     4441 agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     4501 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     4561 tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat
     4621 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
     4681 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     4741 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     4801 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     4861 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     4921 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     4981 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     5041 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     5101 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     5161 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     5221 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     5281 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     5341 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     5401 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     5461 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     5521 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     5581 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     5641 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     5701 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     5761 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     5821 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     5881 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     5941 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     6001 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     6061 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     6121 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     6181 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     6241 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     6301 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     6361 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     6421 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgt
//
