LOCUS       sequence-466365-        4739 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE3107m 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 4739)
  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 4739)
  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: AiP20058 - pAAV-AiE3107m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4739
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     128..146
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(184..206)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     306..325
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(342..797)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(479..500)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     691..710
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   872..1012
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   883..1012
                     /label=AAV2 ITR
     polyA_signal    1130..1241
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     1428..1448
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(1541..2251,2252..2254,2255..2257))
                     /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(1583..1604)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1932..1951
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2191..2212
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2254..2263
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   2884..3013
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(3075..3663)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3155..3174)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3834..4625,4626..4694))
                     /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     4457..4476
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(4695..4739,1..60))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
       61 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
      121 ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga
      181 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
      241 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa
      301 ctatgcggca tcagagcaga ttgtactgag agtgcaccat aaaattgtaa acgttaatat
      361 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aatagaccga
      421 aatcggcaaa atcccttata aatcaaaaga atagcccgag atagagttga gtgttgttcc
      481 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
      541 cgtctatcag ggcgatggcc cactacgtga accatcaccc aaatcaagtt ttttggggtc
      601 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
      661 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctaa
      721 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
      781 gccgctacag ggcgcgtact atggttgctt tgacgtatgc ggtgtgaaat accgcacaga
      841 tgcgtaagga gaaaataccg catcaggcgc ccctgcaggc agctgcgcgc tcgctcgctc
      901 actgaggccg cccgggcaaa gcccgggcgt cgggcgacct ttggtcgccc ggcctcagtg
      961 agcgagcgag cgcgcagaga gggagtggcc aactccatca ctaggggttc ctgcggccgc
     1021 tcggtccgca cgtgagatct catgcctgct attgtcttcc caatcctccc ccttgctgtc
     1081 ctgccccacc ccacccccca gaatagaatg acacctactc agacaatgcg atgcaatttc
     1141 ctcattttat taggaaagga cagtgggagt ggcaccttcc agggtcaagg aaggcacggg
     1201 ggaggggcaa acaacagatg gctggcaact agaaggcaca gtcgaagatc tctcgagcca
     1261 cggaattgtc agtgcccaac agccgagccc ctgtccagca gcgggcaagg caggcggcga
     1321 tgagttccgc cgtggcaaga actaaccagg atttatacaa ggaggagaaa atgaaagcca
     1381 tacgggaagc aatagcatga tacaaaggca ttaaagcagc gtatccacat agcgtaaaag
     1441 gagcaacata gttaagaata ccagtcaatc tttcacaaat tttgtaatcc agaggttgat
     1501 tatgatatcg aattcgcggc cgcggcgcgc cgtcgactta cttgtacagc tcgtccatgc
     1561 cgagagtgat cccggcggcg gtcacgaact ccagcaggac catgtgatcg cgcttctcgt
     1621 tggggtcttt gctcagcttg gactggtagc tcaggtagtg gttgtcgggc agcagcacgg
     1681 ggccgtcgcc gatgggggtg ttctgctggt agtggtcggc gagctgcacg ccgccgtcct
     1741 cgatgttgtg gcggatcttg aagttggcct tgatgccgtt cttctgcttg tcggcggtga
     1801 tatagacgtt gtggctgttg tagttgtact ccagcttgtg ccccaggatg ttgccgtcct
     1861 ccttgaagtc gatgcccttc agctcgatgc ggttcaccag ggtgtcgccc tcgaacttca
     1921 cctcggcgcg ggtcttgtag ttgccgtcgt ccttgaagaa gatggtgcgc tcctggacgt
     1981 agccttcggg catggcggac ttgaagaagt cgtgctgctt catgtggtcg gggtagcggg
     2041 cgaagcactg cacgccgtag cccagggtgg tcacgagggt gggccagggc acgggcagct
     2101 tgccggtggt gcagatcagc ttcagggtca gcttgccgta ggtggcatcg ccctcgccct
     2161 cgccggacac gctgaacttg tggccgttta cgtcgccgtc cagctcgacc aggatgggca
     2221 ccaccccggt gaacagctcc tcgcccttgc tcaccatggt ggcgaccggt agcgctagct
     2281 tcgaaagatc tggatcccag aagcaaatgt aagcaataga tggctctgcc ctgactttta
     2341 tgcccagccc gagctccgcc ggcggtaagg ttttaggaag atgttttctc tgtcttcatc
     2401 tatagggata aagatcaaga agtttcccaa gattcagcct tcttagacca cctgaattct
     2461 ggagttttaa gcattgtact tgttctaaag caacaatcat ctcagcatga gtagttattt
     2521 tgatgggggt ctccatctcc ccttaatttc ttacttatcc actctagatt atctcttgct
     2581 tttatcaaat tgcaactgac ctaatttata aagatttgag tgaaaggaac actcattaaa
     2641 agcctttatc ttttagaagc ctgcctttct tcccccaaat gttctaggtc aatatgatcc
     2701 aagacaaata gtgaggaaga tgtcaggacc actcacaaaa ccttttgaag aaagctggca
     2761 aagagtagaa gctcttaggg tgaggagcaa aggagcagac aagagctgtc caaggatggg
     2821 gctcgtgaca caaggataaa agggcctaag aaatgcctga caaatgtaga cgcgtgcggc
     2881 cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg
     2941 aggccgggcg accaaaggtc gcccgacgcc cgggcggcct cagtgagcga gcgagcgcgc
     3001 agctgcctgc aggacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     3061 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     3121 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     3181 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     3241 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     3301 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     3361 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     3421 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     3481 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     3541 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     3601 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     3661 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     3721 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     3781 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     3841 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     3901 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     3961 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     4021 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     4081 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     4141 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     4201 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     4261 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     4321 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     4381 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     4441 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     4501 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     4561 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     4621 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     4681 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattg
//
