LOCUS       sequence-446833-        5550 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE1179q 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 5550)
  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 5550)
  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: AiP14424 -
            pAAV-AiE1179q-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5550
                     /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      903..912
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(909..911,912..914,915..1625)
                     /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(1215..1234)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1562..1583
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1635..1691
                     /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             1704..1769
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1800..2177
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(2264..2284)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2471..2582
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2700..2840
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2700..2829
                     /label=AAV2 ITR
     misc_feature    2847..2874
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     misc_feature    2875..2902
                     /gene="telL"
                     /label=telL
                     /note="telomeric sequence from coliphage N15"
     rep_origin      2971..3426
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3058..3077)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3268..3289
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3443..3462)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3562..3584
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3622..3640)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3708..3812
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3813..3881,3882..4673)
                     /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(4031..4050)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4844..5432
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5333..5352
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5489..5516
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     misc_feature    5517..5544
                     /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 gcgtatgggc tccagataga aatgatattt ctcagccagc
      181 agggtgtttt ccattacctt aaaatgttca atatttctgc acacattagc tttactaatt
      241 gaagctactt taatgctctc gaaaggttta agaagcaaat gtggaggaac actggtattg
      301 agaagattgt gctgactgat tagcaacgtg tttctgtaaa caggcagcac agtggttggc
      361 ctacctgttg gtgtactatt agaccttgat gaaaaatgtt aattttcatt tgtgttaatg
      421 gctttcgacg ttggtgcttc acacaccgtt tactggcctt ctcagtaggt acggatgcca
      481 gcctccctca ggatggcttt ccatttgcat taagtattca tcacagagat aaatagtcat
      541 ggcctgacac gacacaaatg atggatttga agaaatccaa gcaagtccct ggggatggag
      601 aaaaaggaat gtgaggcttt tgctctcctg agatttgatg gaacctttgc aaagctgctc
      661 acccttttca gatggcatga ctacagacag tctacatctg tttatgtcaa gttgtaaaat
      721 ttacaatgct tatttatata tacattcagt agccagtcat gtagtcaaca catagttttc
      781 ttcagtgact acacatatta ggcacagctg agctcgggct gggcataaaa gtcagggcag
      841 agccatctat tgcttacatt tgcttctggg atccagatct ttcgaagcta gcgctaccgg
      901 tcgccaccat ggtctccaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
      961 agctggacgg cgatgtcaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
     1021 ccacctacgg caagctgacc ctgaagctga tctgcaccac cggcaagctg cccgtgccct
     1081 ggcccaccct cgtgaccacc ctgggctacg gcgtgcagtg cttcgcccgc taccccgacc
     1141 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
     1201 ccatcttctt caaagacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
     1261 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
     1321 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcacc gccgacaagc
     1381 agaagaacgg catcaaggcc aacttcaaga tccgccacaa catcgaggac ggcggcgtgc
     1441 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     1501 acaaccacta cctgagctac cagtccaagc tgagcaaaga ccccaacgag aagcgcgatc
     1561 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
     1621 acaagggctc tggtgctacc aacttctcac tgttgaaaca ggcaggggat gtagaggaga
     1681 atccagggcc tggtgctagt ggagactaca aagaccatga cggagattat aaagatcatg
     1741 acatcgatta caaggatgac gatgacaagt ccggactcag atctggaggc tccggaggcc
     1801 cagagccagc gaagtctgct cccgccccga aaaagggctc caagaaggcg gtgactaagg
     1861 cgcagaagaa aggcggcaag aagcgcaagc gcagccgcaa ggagagctat tccatctatg
     1921 tgtataaggt tctgaagcag gtccaccctg acaccggcat ttcgtccaag gccatgggca
     1981 tcatgaactc gtttgtgaac gacattttcg agcgcatcgc aggtgaggct tcccgcctgg
     2041 cgcattacaa caagcgctcg accatcacct ccagggagat ccagacggcc gtgcgcctgc
     2101 tgctgcctgg ggagttggcc aagcacgccg tgtccgaggg tactaaggcc atcaccaagt
     2161 acaccagcgc taagtaatga ggcgcgccgc ggccgcgaat tcgatatcat aatcaacctc
     2221 tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc
     2281 tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca
     2341 ttttctcctc cttgtataaa tcctggttag ttcttgccac ggcggaactc atcgccgcct
     2401 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggctcgag
     2461 agatcttcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     2521 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     2581 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     2641 ggggaggatt gggaagacaa tagcaggcat gagatctcac gtgcggaccg agcggccgca
     2701 ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc
     2761 cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg
     2821 agcgcgcagc tgcctgcagg ggcgcctatc agcacacaat tgcccattat acgcgcgtat
     2881 aatggactat tgtgtgctga tatgatgcgg tattttctcc ttacgcatct gtgcggtatt
     2941 tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg
     3001 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc
     3061 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     3121 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     3181 ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     3241 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     3301 actctatctc gggctattct tttgatttat aagggatttt gccgatttcg gtctattggt
     3361 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta
     3421 caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc
     3481 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     3541 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     3601 cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     3661 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     3721 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     3781 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     3841 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     3901 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     3961 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     4021 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     4081 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     4141 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     4201 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     4261 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     4321 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     4381 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     4441 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     4501 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     4561 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     4621 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     4681 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     4741 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     4801 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     4861 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     4921 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     4981 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     5041 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     5101 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     5161 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     5221 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     5281 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     5341 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     5401 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     5461 tcctggcctt ttgctggcct tttgctcata tcagcacaca attgcccatt atacgcgcgt
     5521 ataatggact attgtgtgct gataacatgt
//
