LOCUS       sequence-376590-        5423 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AI plasmid ALIAS: AiP13445. Expression of SYFP2 (yellow fluorescent 
            protein) in striatal and lateral septum cholinergic neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5423)
  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 5423)
  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: AiP13445 -
            pAAV-AiE0873m-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5423
                     /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      889..898
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(895..897,898..900,901..1611)
                     /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(1201..1220)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1548..1569
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1621..1677
                     /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             1690..1755
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1786..2163
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(2250..2270)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2457..2568
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2686..2826
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2686..2815
                     /label=AAV2 ITR
     rep_origin      2901..3356
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2988..3007)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3198..3219
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3373..3392)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3492..3514
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3552..3570)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3638..3742
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3743..3811,3812..4603)
                     /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(3961..3980)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4774..5362
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5263..5282
                     /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 gcgtgggctt gagaaatatc atcacaccat tggaagtctt
      181 acaacttgat gtgaacacag agggctttcg gtagaaatat cactgtagga agctttgcag
      241 acaccttcga acccagtgga agtagaagat tcccatgtct gtgttctcta ttcccagcac
      301 cagcatggat ttctccaaag ctttgtaggg cagaactctg aatgtttatt tctctgatga
      361 atccttaatg aaagtggcca gccatcctgg ggagaacaag ccttagttcc cactgagaag
      421 gatagcaagc agcataagag tagctgatgt caaaggctat taatgcaaat gaaaattata
      481 attcctcatc aagatctgct cccagacagg ctaatcattg tgccacctgt ttacagaaac
      541 acactgctaa tcagtcagta aaatctttta agtttctgtg cttctctgtg tacgattttt
      601 agcctctgag agcattaaat taacttcaat tagtaaatct aatggatgct gaagtgttga
      661 gcttttaaat tcaatggaaa acatcctgct ggggaaaaac attatttcta atgagatttt
      721 aagtatttat ttattcattc attcattcat tcattcaaag agggatatat gttaccagtg
      781 catatgtgga ggtcagagct cgggctgggc ataaaagtca gggcagagcc atctattgct
      841 tacatttgct tctgggatcc agatctttcg aagctagcgc taccggtcgc caccatggtc
      901 tccaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgat
      961 gtcaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
     1021 ctgaccctga agctgatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
     1081 accaccctgg gctacggcgt gcagtgcttc gcccgctacc ccgaccacat gaagcagcac
     1141 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaaa
     1201 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     1261 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     1321 gagtacaact acaacagcca caacgtctat atcaccgccg acaagcagaa gaacggcatc
     1381 aaggccaact tcaagatccg ccacaacatc gaggacggcg gcgtgcagct cgccgaccac
     1441 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     1501 agctaccagt ccaagctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     1561 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gggctctggt
     1621 gctaccaact tctcactgtt gaaacaggca ggggatgtag aggagaatcc agggcctggt
     1681 gctagtggag actacaaaga ccatgacgga gattataaag atcatgacat cgattacaag
     1741 gatgacgatg acaagtccgg actcagatct ggaggctccg gaggcccaga gccagcgaag
     1801 tctgctcccg ccccgaaaaa gggctccaag aaggcggtga ctaaggcgca gaagaaaggc
     1861 ggcaagaagc gcaagcgcag ccgcaaggag agctattcca tctatgtgta caaggttctg
     1921 aagcaggtcc accctgacac cggcatttcg tccaaggcca tgggcatcat gaattcgttt
     1981 gtgaacgaca ttttcgagcg catcgcagga gaggcttccc gcctggcgca ttacaacaag
     2041 cgctcgacca tcacctcccg ggagatccag acggccgtgc gcctgctgct gcctggggag
     2101 ttggccaagc acgccgtgtc cgagggtact aaggccatca ccaagtacac cagcgctaag
     2161 taatgaggcg cgccgcggcc gcgaattcga tatcataatc aacctctgga ttacaaaatt
     2221 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     2281 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     2341 tataaatcct ggttagttct tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     2401 tggacagggg ctcggctgtt gggcactgac aattccgtgg ctcgagagat cttcgactgt
     2461 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     2521 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     2581 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     2641 agacaatagc aggcatgaga tctcacgtgc ggaccgagcg gccgcaggaa cccctagtga
     2701 tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg
     2761 tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc
     2821 tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     2881 tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     2941 ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt
     3001 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     3061 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg
     3121 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     3181 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca actctatctc
     3241 gggctattct tttgatttat aagggatttt gccgatttcg gtctattggt taaaaaatga
     3301 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg
     3361 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
     3421 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     3481 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     3541 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     3601 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     3661 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     3721 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     3781 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     3841 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     3901 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     3961 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     4021 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     4081 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     4141 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     4201 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     4261 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     4321 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     4381 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     4441 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     4501 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     4561 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     4621 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     4681 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     4741 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     4801 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     4861 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     4921 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     4981 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     5041 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     5101 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     5161 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     5221 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     5281 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     5341 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     5401 ttgctggcct tttgctcaca tgt
//
