LOCUS       sequence-472835-        4788 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Barcoded enhancer AAV vector for multiplexed screening applications;
            AiE0367h_3xC2 is an enhancer active in striatal MSNs.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4788)
  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 4788)
  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: AiP14449:
            pAAV-AiE0367h_3xC2-minRho-SYFP2-WPRE...
FEATURES             Location/Qualifiers
     source          1..4788
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        15..119
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(120..188,189..980)
                     /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(338..357)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1151..1739
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1640..1659
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   1801..1930
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      2588..2597
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2594..2596,2597..2599,2600..3310)
                     /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(2639..2660)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2900..2919)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3247..3268
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3403..3423)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    3622..3733
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3851..3991
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3851..3980
                     /label=AAV2 ITR
     rep_origin      4066..4521
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4153..4172)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4363..4384
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4538..4557)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4657..4679
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4717..4735)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
       61 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
      121 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
      181 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
      241 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
      301 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
      361 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
      421 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
      481 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
      541 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
      601 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
      661 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
      721 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
      781 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
      841 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
      901 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
      961 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     1021 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     1081 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     1141 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     1201 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     1261 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     1321 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     1381 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     1441 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     1501 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     1561 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     1621 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     1681 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     1741 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     1801 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
     1861 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
     1921 aggggttcct gcggccgcac gcgccggtac cgaagctacc cctaacacac tattctacac
     1981 acagaaaatg ctcttcacta ggaagctacc cctaacacac tattctacac acagaaaatg
     2041 ctcttcacta ggaagctacc cctaacacac tattctacac acagaaaatg ctcttcacta
     2101 gacgcgttcc agcttaaagg aaaattccag ccccagagat cttccgcagt gagtgacagc
     2161 tccttaatgt gaggagcagc gtgggcgtgg tgggtgggat gtcagagcca tcaaagccct
     2221 catttctgat ccctctgtga tgtcggggtt ccctggcacg cccactgtcc agcttaaagg
     2281 aaaattccag ccccagagat cttccgcagt gagtgacagc tccttaatgt gaggagcagc
     2341 gtgggcgtgg tgggtgggat gtcagagcca tcaaagccct catttctgat ccctctgtga
     2401 tgtcggggtt ccctggcacg cccactggag ctcgattcag ccgggagctt agggagggga
     2461 ggtcacttca taagggcttg gggggggagt tggagccacg agtcgtccag ccggagcccc
     2521 gtgtggctgt gctccggcct cagaagcatc cccggatcca gatctttcga agctagcgct
     2581 accggtcgcc accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
     2641 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg
     2701 cgatgccacc tacggcaagc tgaccctgaa gctgatctgc accaccggca agctgcccgt
     2761 gccctggccc accctcgtga ccaccctggg ctacggcgtg cagtgcttcg cccgctaccc
     2821 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
     2881 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
     2941 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
     3001 catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata tcaccgccga
     3061 caagcagaag aacggcatca aggccaactt caagatccgc cacaacatcg aggacggcgg
     3121 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     3181 gcccgacaac cactacctga gctaccagtc caagctgagc aaagacccca acgagaagcg
     3241 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
     3301 gctgtacaag taagtcgacg gcgcgccgcg gccgcgaatt cgatatcata atcaacctct
     3361 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     3421 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     3481 tttctcctcc ttgtataaat cctggttagt tcttgccacg gcggaactca tcgccgcctg
     3541 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggctcgagc
     3601 tagtacgcga gagatcttcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
     3661 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     3721 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     3781 aggacagcaa gggggaggat tgggaagaca atagcaggca tgagatctca cgtgcggacc
     3841 gagcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg
     3901 ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca
     3961 gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg
     4021 catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg
     4081 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     4141 ccttagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     4201 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     4261 tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga
     4321 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     4381 ctggaacaac actcaactct atctcgggct attcttttga tttataaggg attttgccga
     4441 tttcggtcta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     4501 aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat
     4561 agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     4621 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     4681 tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat
     4741 aggttaatgt catgataata atggtttctt agacgtcagg tggcactt
//
