LOCUS       sequence-466367-        4718 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE3111m 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 4718)
  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 4718)
  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: AiP20060 - pAAV-AiE3111m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4718
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     279..298
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(517..621)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     689..707
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(745..767)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     867..886
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(903..1358)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1040..1061)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1252..1271
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   1433..1573
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1444..1573
                     /label=AAV2 ITR
     polyA_signal    1691..1802
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     1989..2009
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(2102..2812,2813..2815,2816..2818))
                     /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(2144..2165)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2493..2512
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2752..2773
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2815..2824
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   3424..3553
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(3615..4203)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3695..3714)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4374..4718,1..447,448..516))
                     /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"
ORIGIN
        1 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
       61 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
      121 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
      181 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
      241 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
      301 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
      361 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
      421 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
      481 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
      541 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
      601 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
      661 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc
      721 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
      781 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
      841 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
      901 taaaattgta aacgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
      961 attttttaac caatagaccg aaatcggcaa aatcccttat aaatcaaaag aatagcccga
     1021 gatagagttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     1081 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     1141 caaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     1201 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     1261 agcgaaagga gcgggcgcta aggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     1321 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtac tatggttgct ttgacgtatg
     1381 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg cccctgcagg
     1441 cagctgcgcg ctcgctcgct cactgaggcc gcccgggcaa agcccgggcg tcgggcgacc
     1501 tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc
     1561 actaggggtt cctgcggccg ctcggtccgc acgtgagatc tcatgcctgc tattgtcttc
     1621 ccaatcctcc cccttgctgt cctgccccac cccacccccc agaatagaat gacacctact
     1681 cagacaatgc gatgcaattt cctcatttta ttaggaaagg acagtgggag tggcaccttc
     1741 cagggtcaag gaaggcacgg gggaggggca aacaacagat ggctggcaac tagaaggcac
     1801 agtcgaagat ctctcgagcc acggaattgt cagtgcccaa cagccgagcc cctgtccagc
     1861 agcgggcaag gcaggcggcg atgagttccg ccgtggcaag aactaaccag gatttataca
     1921 aggaggagaa aatgaaagcc atacgggaag caatagcatg atacaaaggc attaaagcag
     1981 cgtatccaca tagcgtaaaa ggagcaacat agttaagaat accagtcaat ctttcacaaa
     2041 ttttgtaatc cagaggttga ttatgatatc gaattcgcgg ccgcggcgcg ccgtcgactt
     2101 acttgtacag ctcgtccatg ccgagagtga tcccggcggc ggtcacgaac tccagcagga
     2161 ccatgtgatc gcgcttctcg ttggggtctt tgctcagctt ggactggtag ctcaggtagt
     2221 ggttgtcggg cagcagcacg gggccgtcgc cgatgggggt gttctgctgg tagtggtcgg
     2281 cgagctgcac gccgccgtcc tcgatgttgt ggcggatctt gaagttggcc ttgatgccgt
     2341 tcttctgctt gtcggcggtg atatagacgt tgtggctgtt gtagttgtac tccagcttgt
     2401 gccccaggat gttgccgtcc tccttgaagt cgatgccctt cagctcgatg cggttcacca
     2461 gggtgtcgcc ctcgaacttc acctcggcgc gggtcttgta gttgccgtcg tccttgaaga
     2521 agatggtgcg ctcctggacg tagccttcgg gcatggcgga cttgaagaag tcgtgctgct
     2581 tcatgtggtc ggggtagcgg gcgaagcact gcacgccgta gcccagggtg gtcacgaggg
     2641 tgggccaggg cacgggcagc ttgccggtgg tgcagatcag cttcagggtc agcttgccgt
     2701 aggtggcatc gccctcgccc tcgccggaca cgctgaactt gtggccgttt acgtcgccgt
     2761 ccagctcgac caggatgggc accaccccgg tgaacagctc ctcgcccttg ctcaccatgg
     2821 tggcgaccgg tagcgctagc ttcgaaagat ctggatccca gaagcaaatg taagcaatag
     2881 atggctctgc cctgactttt atgcccagcc cgagctccgc cggcgatcag tgagatgctt
     2941 gggaggcttg cagaagggct gggaaggtgg agagagacgt cagcaagatg aacaggacgg
     3001 ctggcgacgt atgcaggaga tgctagtaag aggtaagctt cctattctga gggataatga
     3061 catcagcctt cagggaagcc acagctgaga atcctagaaa gatgaaggcc cactggcacc
     3121 tgaaaaaaaa agaaagaaaa gaaattgcag aaaaaaagca tccgagcctc aaatcttctc
     3181 cattctgtta aatcctgctg gaggcagata cagcagccgc gagaccagct gattccagga
     3241 aactgtcaaa aggaggaatc accctcccta agcagcaaga aagccacatc ccagcagggc
     3301 agaaacccag ccaggtgtca ctgagaaggc ctggcaatgg ctacagaagc caggctgttc
     3361 gtgggtatcg atcctcactg tctctctttg tccctccaag tgattccaaa cgcgtgcggc
     3421 cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg
     3481 aggccgggcg accaaaggtc gcccgacgcc cgggcggcct cagtgagcga gcgagcgcgc
     3541 agctgcctgc aggacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     3601 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     3661 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     3721 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     3781 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     3841 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     3901 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     3961 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     4021 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     4081 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     4141 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     4201 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     4261 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     4321 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     4381 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     4441 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     4501 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     4561 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     4621 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     4681 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtt
//
