LOCUS       sequence-470438-        4442 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0372m is an enhancer sequence, designed to drive AAV-mediated 
            transgene expression in specific populations of brain cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4442)
  AUTHORS   Ben-Simon Y, Hooper M, Narayan S, Daigle TL, Dwivedi D, Way SW, 
            Oster A, Stafford DA, Mich JK, Taormina MJ, Martinez RA, Opitz-Araya
            X, Roth JR, Alexander JR, Allen S, Amster A, Arbuckle J, Ayala A, 
            Baker PM, Bakken TE, Barcelli T, Barta S, Bendrick J, Bertagnolli D,
            Bielstein C, Bishwakarma P, Bowlus J, Boyer G, Brouner K, Casian B, 
            Casper T, Chakka AB, Chakrabarty R, Chance RK, Chavan S, Clark M, 
            Colbert K, Collman F, Daniel S, Departee M, DiValentin P, Donadio N,
            Dotson N, Egdorf T, Fliss T, Gabitto M, Garcia J, Gary A, Gasperini 
            M, Gloe J, Goldy J, Gore BB, Graybuck L, Greisman N, Haeseleer F, 
            Halterman C, Haradon Z, Hastings SD, Helback O, Ho W, Hockemeyer D, 
            Huang C, Huff S, Hunker A, Johansen N, Jones D, Juneau Z, Kalmbach 
            B, Kannan M, Khem S, Kussick E, Kutsal R, Larsen R, Lee C, Lee AY, 
            Leibly M, Lenz GH, Li S, Liang E, Lusk N, Madigan Z, Malloy J, 
            Malone J, McCue R, Melchor J, Mollenkopf T, Moosman S, Morin E, 
            Newman D, Ng L, Ngo K, Omstead V, Otto S, Oyama A, Pena N, Pham T, 
            Phillips E, Pom CA, Potekhina L, Ransford S, Ray PL, Rette D, 
            Reynoldson C, Rimorin C, Rocha D, Ruiz A, Sanchez REA, Sawyer L, 
            Sedeno-Cortes A, Sevigny JP, Shapovalova N, Shepard N, Shulga L, 
            Sigler AR, Siverts L, Soliman S, Somasundaram S, Staats B, Stewart 
            K, Szelenyi E, Tieu M, Trader C, Tran A, van Velthoven CTJ, Walker 
            M, Wang Y, Weed N, Wirthlin M, Wood T, Wynalda B, Yao Z, Zhou T, 
            Ariza J, Dee N, Reding M, Ronellenfitch K, Mufti S, Sunkin SM, Smith
            KA, Esposito L, Waters J, Thyagarajan B, Yao S, Lein ES, Zeng H, 
            Levi BP, Ngai J, Ting JT, Tasic B
  TITLE     A suite of enhancer AAVs and transgenic mouse lines for genetic 
            access to cortical cell types.
  JOURNAL   Cell. 2025 May 16:S0092-8674(25)00513-6. doi: 
            10.1016/j.cell.2025.05.002.
  PUBMED    40403729
REFERENCE   2  (bases 1 to 4442)
  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"; journalName: "Cell. 2025 
            May 16:S0092-8674(25)00513-6. doi: 10.1016/j.cell.2025.05.002."
COMMENT     Sequence Label: AiP12083 - pAAV-AiE0372m-minBG-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..4442
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(146..734)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(226..245)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(905..1696,1697..1765))
                     /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     1528..1547
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1766..1870)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1938..1956
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1994..2016)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2116..2135
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2152..2607)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2289..2310)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2501..2520
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2682..2822
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2693..2822
                     /label=AAV2 ITR
     polyA_signal    2940..3051
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3238..3258
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(3351..4061,4062..4064,4065..4067))
                     /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(3393..3414)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3742..3761
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4001..4022
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4064..4073
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   join(4397..4442,1..84)
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggcggcc tcagtgagcg
       61 agcgagcgcg cagctgcctg caggacatgt gagcaaaagg ccagcaaaag gccaggaacc
      121 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
      181 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
      241 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
      301 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
      361 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
      421 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
      481 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
      541 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
      601 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
      661 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
      721 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
      781 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
      841 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
      901 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
      961 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     1021 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     1081 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     1141 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     1201 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     1261 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     1321 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     1381 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     1441 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     1501 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     1561 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     1621 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     1681 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     1741 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     1801 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     1861 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
     1921 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg
     1981 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     2041 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     2101 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat aaaattgtaa
     2161 acgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     2221 aatagaccga aatcggcaaa atcccttata aatcaaaaga atagcccgag atagagttga
     2281 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     2341 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc aaatcaagtt
     2401 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     2461 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     2521 cgggcgctaa ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     2581 cgcttaatgc gccgctacag ggcgcgtact atggttgctt tgacgtatgc ggtgtgaaat
     2641 accgcacaga tgcgtaagga gaaaataccg catcaggcgc ccctgcaggc agctgcgcgc
     2701 tcgctcgctc actgaggccg cccgggcaaa gcccgggcgt cgggcgacct ttggtcgccc
     2761 ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc aactccatca ctaggggttc
     2821 ctgcggccgc tcggtccgca cgtgagatct catgcctgct attgtcttcc caatcctccc
     2881 ccttgctgtc ctgccccacc ccacccccca gaatagaatg acacctactc agacaatgcg
     2941 atgcaatttc ctcattttat taggaaagga cagtgggagt ggcaccttcc agggtcaagg
     3001 aaggcacggg ggaggggcaa acaacagatg gctggcaact agaaggcaca gtcgaagatc
     3061 tctcgagcca cggaattgtc agtgcccaac agccgagccc ctgtccagca gcgggcaagg
     3121 caggcggcga tgagttccgc cgtggcaaga actaaccagg atttatacaa ggaggagaaa
     3181 atgaaagcca tacgggaagc aatagcatga tacaaaggca ttaaagcagc gtatccacat
     3241 agcgtaaaag gagcaacata gttaagaata ccagtcaatc tttcacaaat tttgtaatcc
     3301 agaggttgat tatgatatcg aattcgcggc cgcggcgcgc cgtcgactta cttgtacagc
     3361 tcgtccatgc cgagagtgat cccggcggcg gtcacgaact ccagcaggac catgtgatcg
     3421 cgcttctcgt tggggtcttt gctcagcttg gactggtagc tcaggtagtg gttgtcgggc
     3481 agcagcacgg ggccgtcgcc gatgggggtg ttctgctggt agtggtcggc gagctgcacg
     3541 ccgccgtcct cgatgttgtg gcggatcttg aagttggcct tgatgccgtt cttctgcttg
     3601 tcggcggtga tatagacgtt gtggctgttg tagttgtact ccagcttgtg ccccaggatg
     3661 ttgccgtcct ccttgaagtc gatgcccttc agctcgatgc ggttcaccag ggtgtcgccc
     3721 tcgaacttca cctcggcgcg ggtcttgtag ttgccgtcgt ccttgaagaa gatggtgcgc
     3781 tcctggacgt agccttcggg catggcggac ttgaagaagt cgtgctgctt catgtggtcg
     3841 gggtagcggg cgaagcactg cacgccgtag cccagggtgg tcacgagggt gggccagggc
     3901 acgggcagct tgccggtggt gcagatcagc ttcagggtca gcttgccgta ggtggcatcg
     3961 ccctcgccct cgccggacac gctgaacttg tggccgttta cgtcgccgtc cagctcgacc
     4021 aggatgggca ccaccccggt gaacagctcc tcgcccttgc tcaccatggt ggcgaccggt
     4081 agcgctagct tcgaaagatc tggatcccag aagcaaatgt aagcaataga tggctctgcc
     4141 ctgactttta tgcccagccc gagctcaagg cactgtggca taccctatgc tgggctttat
     4201 tcatcaggga gaaaggtatc cccttttatt catggatctg caacaagact tagtgtcatt
     4261 tcatccaatt ttcatagaat ggccagtcca ctcggcaggg ctatgtaaaa taagaagaaa
     4321 tctccagggg gtgtttttat atgtggaacc agcagagtcc tggcacgtta ggaagcaaac
     4381 gaacgcgtgc ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct
     4441 cg
//
