LOCUS       sequence-446822-        5546 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE1180h is an enhancer sequence, designed to drive AAV-mediated 
            transgene expression in striatal cholinergic neurons (weak).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5546)
  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 5546)
  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: AiP14425 -
            pAAV-AiE1180h-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5546
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      899..908
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(905..907,908..910,911..1621)
                     /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(1211..1230)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1558..1579
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1631..1687
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             1700..1765
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1796..2173
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(2260..2280)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2467..2578
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2696..2836
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2696..2825
                     /label=AAV2 ITR
     misc_feature    2843..2870
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     misc_feature    2871..2898
                     /gene="telL"
                     /label=telL
                     /note="telomeric sequence from coliphage N15"
     rep_origin      2967..3422
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3054..3073)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3264..3285
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3439..3458)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3558..3580
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3618..3636)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3704..3808
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3809..3877,3878..4669)
                     /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(4027..4046)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4840..5428
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5329..5348
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5485..5512
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     misc_feature    5513..5540
                     /gene="telL"
                     /label=telL
                     /note="telomeric sequence from coliphage N15"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtggctcc agatagaaat gatatttctc acccagcagg
      181 gtgttttcca ttaaattaaa atgttcaata tttctgcaca cattagcttt actaattgaa
      241 actaatttaa tgctctcaaa aagcttaaga agcaaatgtg gaggaacact ggtattgagt
      301 ggattgtgct gactgattag caacgtgttt ctgtaaacag gcagcacagt ggttggccta
      361 cctgttggtg tactgttaga ccttgatgaa aaatgtcaat ttgcatttgc attaatggcc
      421 tttgacgtcg gtgcttcaca cactgtttgc tggccttctc agtaggtacg gatgccagcc
      481 tccctcagga tggcttgcca tttgcattaa gtatttatca cagagagaaa tagtcatggc
      541 ctggcacaac acaaacgatg gatttggaga aatccaagca agtccctggg gatagagaaa
      601 aaggaatgtg aggcttttgc tttcctgaga tttgatggaa tctttgaaaa gctgctcacc
      661 cttttcagtt ggcatgacta caagtagtct acatttgttt atgtcaagtt gtaaaattta
      721 ccatgcctat ttatatatac attcagtagc cagtcacgta gtcaacaaat agttttgttc
      781 agtgactaca catattagac acagcgagct cgggctgggc ataaaagtca gggcagagcc
      841 atctattgct tacatttgct tctgggatcc agatctttcg aagctagcgc taccggtcgc
      901 caccatggtc tccaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
      961 ggacggcgat gtcaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
     1021 ctacggcaag ctgaccctga agctgatctg caccaccggc aagctgcccg tgccctggcc
     1081 caccctcgtg accaccctgg gctacggcgt gcagtgcttc gcccgctacc ccgaccacat
     1141 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
     1201 cttcttcaaa gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     1261 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     1321 gcacaagctg gagtacaact acaacagcca caacgtctat atcaccgccg acaagcagaa
     1381 gaacggcatc aaggccaact tcaagatccg ccacaacatc gaggacggcg gcgtgcagct
     1441 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     1501 ccactacctg agctaccagt ccaagctgag caaagacccc aacgagaagc gcgatcacat
     1561 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     1621 gggctctggt gctaccaact tctcactgtt gaaacaggca ggggatgtag aggagaatcc
     1681 agggcctggt gctagtggag actacaaaga ccatgacgga gattataaag atcatgacat
     1741 cgattacaag gatgacgatg acaagtccgg actcagatct ggaggctccg gaggcccaga
     1801 gccagcgaag tctgctcccg ccccgaaaaa gggctccaag aaggcggtga ctaaggcgca
     1861 gaagaaaggc ggcaagaagc gcaagcgcag ccgcaaggag agctattcca tctatgtgta
     1921 taaggttctg aagcaggtcc accctgacac cggcatttcg tccaaggcca tgggcatcat
     1981 gaactcgttt gtgaacgaca ttttcgagcg catcgcaggt gaggcttccc gcctggcgca
     2041 ttacaacaag cgctcgacca tcacctccag ggagatccag acggccgtgc gcctgctgct
     2101 gcctggggag ttggccaagc acgccgtgtc cgagggtact aaggccatca ccaagtacac
     2161 cagcgctaag taatgaggcg cgccgcggcc gcgaattcga tatcataatc aacctctgga
     2221 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     2281 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     2341 ctcctccttg tataaatcct ggttagttct tgccacggcg gaactcatcg ccgcctgcct
     2401 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg ctcgagagat
     2461 cttcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
     2521 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
     2581 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
     2641 aggattggga agacaatagc aggcatgaga tctcacgtgc ggaccgagcg gccgcaggaa
     2701 cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg
     2761 cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg
     2821 cgcagctgcc tgcaggggcg cctatcagca cacaattgcc cattatacgc gcgtataatg
     2881 gactattgtg tgctgatatg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     2941 accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa gcgcggcggg
     3001 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccttagcgc ccgctccttt
     3061 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     3121 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     3181 tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     3241 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaactc
     3301 tatctcgggc tattcttttg atttataagg gattttgccg atttcggtct attggttaaa
     3361 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat
     3421 tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca
     3481 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
     3541 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
     3601 acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     3661 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     3721 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     3781 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     3841 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     3901 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     3961 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     4021 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     4081 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     4141 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     4201 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     4261 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     4321 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     4381 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     4441 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     4501 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     4561 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     4621 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     4681 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     4741 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     4801 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     4861 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     4921 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     4981 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     5041 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     5101 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     5161 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     5221 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     5281 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     5341 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     5401 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     5461 ggccttttgc tggccttttg ctcatatcag cacacaattg cccattatac gcgcgtataa
     5521 tggactattg tgtgctgata acatgt
//
