LOCUS       sequence-446782-        5607 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 5607)
  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 5607)
  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: AiP15355 - pAAV-AiE0784m-minBG-DIO-SYFP2-WPRE3...
FEATURES             Location/Qualifiers
     source          1..5607
                     /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(853..1563,1564..1566,1567..1569))
                     /codon_start=1
                     /product="monomeric optimized variant of EYFP, identical to
                     mVenus but with L68V (Kremers et al., 2006)"
                     /label=SYFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(895..916)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1244..1263
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1503..1524
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1566..1575
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    1592..1625
                     /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    1676..1709
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    1734..2322
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1787..1807)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2205..2216)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2354..2830
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2553..2572)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   2870..3010
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2870..2999
                     /label=AAV2 ITR
     rep_origin      3085..3540
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3172..3191)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3382..3403
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3557..3576)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3676..3698
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3736..3754)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3822..3926
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3927..3995,3996..4787)
                     /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(4145..4164)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4958..5546
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5447..5466
                     /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 gccgtcgact tacttgtaca gctcgtccat gccgagagtg atcccggcgg cggtcacgaa
      901 ctccagcagg accatgtgat cgcgcttctc gttggggtct ttgctcagct tggactggta
      961 gctcaggtag tggttgtcgg gcagcagcac ggggccgtcg ccgatggggg tgttctgctg
     1021 gtagtggtcg gcgagctgca cgccgccgtc ctcgatgttg tggcggatct tgaagttggc
     1081 cttgatgccg ttcttctgct tgtcggcggt gatatagacg ttgtggctgt tgtagttgta
     1141 ctccagcttg tgccccagga tgttgccgtc ctccttgaag tcgatgccct tcagctcgat
     1201 gcggttcacc agggtgtcgc cctcgaactt cacctcggcg cgggtcttgt agttgccgtc
     1261 gtccttgaag aagatggtgc gctcctggac gtagccttcg ggcatggcgg acttgaagaa
     1321 gtcgtgctgc ttcatgtggt cggggtagcg ggcgaagcac tgcacgccgt agcccagggt
     1381 ggtcacgagg gtgggccagg gcacgggcag cttgccggtg gtgcagatca gcttcagggt
     1441 cagcttgccg taggtggcat cgccctcgcc ctcgccggac acgctgaact tgtggccgtt
     1501 tacgtcgccg tccagctcga ccaggatggg caccaccccg gtgaacagct cctcgccctt
     1561 gctcaccatg gtggcgaccg gtagcgctag cataacttcg tataaagtat cctatacgaa
     1621 gttatttgcc ttaacccaga aattatcact gttattcttt agaatggtgc aaagaataac
     1681 ttcgtataat gtatgctata cgaagttatg aattcgatat caagcttatc gataatcaac
     1741 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
     1801 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
     1861 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg
     1921 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg
     1981 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca
     2041 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca
     2101 ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctatg
     2161 ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag
     2221 cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc
     2281 gccctcagac gagtcggatc tccctttggg ccgcctcccc gcatcgatac cgagcgctgc
     2341 tcgagagatc tacgggtggc atccctgtga cccctcccca gtgcctctcc tggccctgga
     2401 agttgccact ccagtgccca ccagccttgt cctaataaaa ttaagttgca tcattttgtc
     2461 tgactaggtg tccttctata atattatggg gtggaggggg gtggtatgga gcaaggggca
     2521 agttgggaag acaacctgta gggcctgcgg ggtctattgg gaaccaagct ggagtgcagt
     2581 ggcacaatct tggctcactg caatctccgc ctcctgggtt caagcgattc tcctgcctca
     2641 gcctcccgag ttgttgggat tccaggcatg catgaccagg ctcagctaat ttttgttttt
     2701 ttggtagaga cggggtttca ccatattggc caggctggtc tccaactcct aatctcaggt
     2761 gatctaccca ccttggcctc ccaaattgct gggattacag gcgtgaacca ctgctccctt
     2821 ccctgtcctt ctgattttgt aggtaaccac gtgcggaccg agcggccgca ggaaccccta
     2881 gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca
     2941 aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc
     3001 tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     3061 cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     3121 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cttagcgccc gctcctttcg
     3181 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     3241 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     3301 tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     3361 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaactcta
     3421 tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggtctat tggttaaaaa
     3481 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt
     3541 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc
     3601 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
     3661 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
     3721 gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
     3781 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     3841 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     3901 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     3961 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     4021 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     4081 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     4141 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     4201 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     4261 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     4321 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     4381 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     4441 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     4501 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     4561 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     4621 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     4681 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     4741 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     4801 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     4861 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     4921 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     4981 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     5041 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     5101 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     5161 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     5221 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     5281 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     5341 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     5401 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     5461 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     5521 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     5581 ccttttgctg gccttttgct cacatgt
//
