LOCUS       sequence-459487-        5984 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0779m_3xC2 is an enhancer sequence, designed to drive 
            AAV-mediated transgene expression in striatal direct pathway MSNs 
            (D1-MSNs).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5984)
  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 5984)
  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: AiP14210:
            pAAV-AiE0779m_3xC2-minBG-SYFP2-P2A-m...
FEATURES             Location/Qualifiers
     source          1..5984
                     /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      820..829
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(826..828,829..831,832..1542)
                     /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(871..892)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1132..1151)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1479..1500
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1552..1608
                     /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             1624..2319
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(1757..1775)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2025..2044
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             2350..2727
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(2811..2831)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    3018..3129
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3247..3387
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3247..3376
                     /label=AAV2 ITR
     rep_origin      3462..3917
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3549..3568)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3759..3780
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3934..3953)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4053..4075
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4113..4131)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4199..4303
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4304..4372,4373..5164)
                     /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(4522..4541)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5335..5923
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5824..5843
                     /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 gcgtaagttt gagacaaatt aaggaaaagc gggttgggag
      181 gagtctgagg ctgtgggtgg ctgcctcgag aggctgctca caacctgaca tgtttcagat
      241 cttttgacac ggtcatgcaa gcaacagaaa cttatatggt ttttagtctc ccaggtagca
      301 gggaaatgat gatgtgggag gattaggggg gggaaaataa gtttgagaca aattaaggaa
      361 aagcgggttg ggaggagtct gaggctgtgg gtggctgcct cgagaggctg ctcacaacct
      421 gacatgtttc agatcttttg acacggtcat gcaagcaaca gaaacttata tggtttttag
      481 tctcccaggt agcagggaaa tgatgatgtg ggaggattag gggggggaaa ataagtttga
      541 gacaaattaa ggaaaagcgg gttgggagga gtctgaggct gtgggtggct gcctcgagag
      601 gctgctcaca acctgacatg tttcagatct tttgacacgg tcatgcaagc aacagaaact
      661 tatatggttt ttagtctccc aggtagcagg gaaatgatga tgtgggagga ttaggggggg
      721 gaaaatgagc tcgggctggg cataaaagtc agggcagagc catctattgc ttacatttgc
      781 ttctgggatc cagatctttc gaagctagcg ctaccggtcg ccaccatggt gagcaagggc
      841 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
      901 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
      961 aagctgatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
     1021 ggctacggcg tgcagtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc
     1081 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
     1141 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     1201 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     1261 tacaacagcc acaacgtcta tatcaccgcc gacaagcaga agaacggcat caaggccaac
     1321 ttcaagatcc gccacaacat cgaggacggc ggcgtgcagc tcgccgacca ctaccagcag
     1381 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagctaccag
     1441 tccaagctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
     1501 accgccgccg ggatcactct cggcatggac gagctgtaca agggaagcgg agccactaac
     1561 ttctccctgt tgaaacaagc aggggatgtc gaagagaatc ccgggccagc accgggctcc
     1621 accatggtga gcaagggcga ggcagtgatc aaggagttca tgcggttcaa ggtgcacatg
     1681 gagggctcca tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccctac
     1741 gagggcaccc agaccgccaa gctgaaggtg accaagggtg gccccctgcc cttctcctgg
     1801 gacatcctgt cccctcagtt catgtacggc tccagggcct tcaccaagca ccccgccgac
     1861 atccccgact actataagca gtccttcccc gagggcttca agtgggagcg cgtgatgaac
     1921 ttcgaggacg gcggcgccgt gaccgtgacc caggacacct ccctggagga cggcaccctg
     1981 atctacaagg tgaagctccg cggcaccaac ttccctcctg acggccccgt aatgcagaag
     2041 aagacaatgg gctgggaagc gtccaccgag cggttgtacc ccgaggacgg cgtgctgaag
     2101 ggcgacatta agatggccct gcgcctgaag gacggcggcc gctacctggc ggacttcaag
     2161 accacctaca aggccaagaa gcccgtgcag atgcccggcg cctacaacgt cgaccgcaag
     2221 ttggacatca cctcccacaa cgaggactac accgtggtgg aacagtacga acgctccgag
     2281 ggccgccact ccaccggcgg catggacgag ctgtataagt ccggactcag atctggaggc
     2341 tccggaggcc cagagccagc gaagtctgct cccgccccga aaaagggctc caagaaggcg
     2401 gtgactaagg cgcagaagaa aggcggcaag aagcgcaagc gcagccgcaa ggagagctat
     2461 tccatctatg tgtataaggt tctgaagcag gtccaccctg acaccggcat ttcgtccaag
     2521 gccatgggca tcatgaactc gtttgtgaac gacattttcg agcgcatcgc aggtgaggct
     2581 tcccgcctgg cgcattacaa caagcgctcg accatcacct ccagggagat ccagacggcc
     2641 gtgcgcctgc tgctgcctgg ggagttggcc aagcacgccg tgtccgaggg tactaaggcc
     2701 atcaccaagt acaccagcgc taagtaaggc gcgcccctgc agggaattcg atatcataat
     2761 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     2821 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     2881 gctttcattt tctcctcctt gtataaatcc tggttagttc ttgccacggc ggaactcatc
     2941 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     3001 gctcgagaga tcttcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     3061 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     3121 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     3181 cagcaagggg gaggattggg aagacaatag caggcatgag atctcacgtg cggaccgagc
     3241 ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca
     3301 ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga
     3361 gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc cttacgcatc
     3421 tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc
     3481 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgcctt
     3541 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     3601 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     3661 ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     3721 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     3781 aacaacactc aactctatct cgggctattc ttttgattta taagggattt tgccgatttc
     3841 ggtctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     3901 attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     3961 aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     4021 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     4081 accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt
     4141 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     4201 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     4261 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     4321 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     4381 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     4441 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     4501 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     4561 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     4621 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     4681 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     4741 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     4801 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     4861 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     4921 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     4981 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     5041 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     5101 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     5161 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     5221 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     5281 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     5341 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     5401 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     5461 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     5521 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     5581 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     5641 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     5701 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     5761 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     5821 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     5881 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     5941 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgt
//
