LOCUS       sequence-466364-        4964 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE3106m 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 4964)
  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 4964)
  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: AiP20057 - pAAV-AiE3106m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4964
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   216..345
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(407..995)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(487..506)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1166..1957,1958..2026))
                     /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     1789..1808
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2027..2131)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2199..2217
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2255..2277)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2377..2396
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2413..2868)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2550..2571)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2762..2781
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2943..3083
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2954..3083
                     /label=AAV2 ITR
     polyA_signal    3201..3312
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3499..3519
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(3612..4322,4323..4325,4326..4328))
                     /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(3654..3675)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4003..4022
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4262..4283
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4325..4334
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 gcttgcagag tagaagagag gcagagttgg tgagttcctt cttcctctgc tcataggttt
       61 accagagaag gtcccagatc tgcaccccaa ccctcaatcc ctcttcaggc accctctccc
      121 tgggatgctt cactctccca gcagcccctg cctggatact aatgatgcca agtgcttgag
      181 caccaatctc gactttgggg cacgcgtgcg gccgcaggaa cccctagtga tggagttggc
      241 cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg
      301 cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggacatg tgagcaaaag
      361 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
      421 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
      481 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
      541 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
      601 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
      661 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
      721 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
      781 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
      841 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
      901 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
      961 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     1021 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     1081 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     1141 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     1201 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     1261 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     1321 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     1381 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     1441 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     1501 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     1561 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     1621 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     1681 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     1741 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     1801 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     1861 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     1921 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     1981 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     2041 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     2101 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     2161 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
     2221 ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga
     2281 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
     2341 cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga
     2401 gagtgcacca taaaattgta aacgttaata ttttgttaaa attcgcgtta aatttttgtt
     2461 aaatcagctc attttttaac caatagaccg aaatcggcaa aatcccttat aaatcaaaag
     2521 aatagcccga gatagagttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     2581 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     2641 aaccatcacc caaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     2701 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     2761 aagggaagaa agcgaaagga gcgggcgcta aggcgctggc aagtgtagcg gtcacgctgc
     2821 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtac tatggttgct
     2881 ttgacgtatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
     2941 cccctgcagg cagctgcgcg ctcgctcgct cactgaggcc gcccgggcaa agcccgggcg
     3001 tcgggcgacc tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc
     3061 caactccatc actaggggtt cctgcggccg ctcggtccgc acgtgagatc tcatgcctgc
     3121 tattgtcttc ccaatcctcc cccttgctgt cctgccccac cccacccccc agaatagaat
     3181 gacacctact cagacaatgc gatgcaattt cctcatttta ttaggaaagg acagtgggag
     3241 tggcaccttc cagggtcaag gaaggcacgg gggaggggca aacaacagat ggctggcaac
     3301 tagaaggcac agtcgaagat ctctcgagcc acggaattgt cagtgcccaa cagccgagcc
     3361 cctgtccagc agcgggcaag gcaggcggcg atgagttccg ccgtggcaag aactaaccag
     3421 gatttataca aggaggagaa aatgaaagcc atacgggaag caatagcatg atacaaaggc
     3481 attaaagcag cgtatccaca tagcgtaaaa ggagcaacat agttaagaat accagtcaat
     3541 ctttcacaaa ttttgtaatc cagaggttga ttatgatatc gaattcgcgg ccgcggcgcg
     3601 ccgtcgactt acttgtacag ctcgtccatg ccgagagtga tcccggcggc ggtcacgaac
     3661 tccagcagga ccatgtgatc gcgcttctcg ttggggtctt tgctcagctt ggactggtag
     3721 ctcaggtagt ggttgtcggg cagcagcacg gggccgtcgc cgatgggggt gttctgctgg
     3781 tagtggtcgg cgagctgcac gccgccgtcc tcgatgttgt ggcggatctt gaagttggcc
     3841 ttgatgccgt tcttctgctt gtcggcggtg atatagacgt tgtggctgtt gtagttgtac
     3901 tccagcttgt gccccaggat gttgccgtcc tccttgaagt cgatgccctt cagctcgatg
     3961 cggttcacca gggtgtcgcc ctcgaacttc acctcggcgc gggtcttgta gttgccgtcg
     4021 tccttgaaga agatggtgcg ctcctggacg tagccttcgg gcatggcgga cttgaagaag
     4081 tcgtgctgct tcatgtggtc ggggtagcgg gcgaagcact gcacgccgta gcccagggtg
     4141 gtcacgaggg tgggccaggg cacgggcagc ttgccggtgg tgcagatcag cttcagggtc
     4201 agcttgccgt aggtggcatc gccctcgccc tcgccggaca cgctgaactt gtggccgttt
     4261 acgtcgccgt ccagctcgac caggatgggc accaccccgg tgaacagctc ctcgcccttg
     4321 ctcaccatgg tggcgaccgg tagcgctagc ttcgaaagat ctggatccca gaagcaaatg
     4381 taagcaatag atggctctgc cctgactttt atgcccagcc cgagctccgc cggcgttata
     4441 attacattgg aagaccccag gcaaccaagt aaactgatta gctgcttctg cagagttgat
     4501 ggattttccc cctccctctg ggtcacaaat agggccttgg gagatggggg cagccctgta
     4561 ggtgggagtc attcaggatg gggctatcta tatccaggaa gtcaaactga gtggtactga
     4621 caggttccct tggagcctct gtctgacagg tgcccacttg cccccagaac aggctgcccg
     4681 ggctccctcc cagccaggtt gtccgttgat cctgagcaga tcaagagcca ggcagaacag
     4741 gctggtggtg gctggaaagc cctcccccag gagagccgaa gcgttctgcc aggcataatt
     4801 attgtatgga taattacaca attcggagcc tgctggaaaa ccctgagctg gagccagagc
     4861 atggaagcag tgttctggga ctatggccgc ttgctgcttc tcagagttga gtttaaaaga
     4921 ggataacccc tccccctcct agaactccaa tgcccttgct gatg
//
