LOCUS       sequence-466349-        4785 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE3056m 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 4785)
  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 4785)
  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: AiP20019 - pAAV-AiE3056m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4785
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      415..424
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(421..423,424..426,427..1137)
                     /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(466..487)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(727..746)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1074..1095
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1230..1250)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1437..1548
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1666..1806
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1666..1795
                     /label=AAV2 ITR
     rep_origin      1881..2336
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1968..1987)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2178..2199
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2353..2372)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2472..2494
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2532..2550)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2618..2722
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2723..2791,2792..3583)
                     /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(2941..2960)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3754..4342
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4243..4262
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   4404..4533
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 ggctttaaat gacatgatca gatttgcatt ttaataagag ctcagtggct gctgtggaga
       61 gcatgaacca cagtgcgggt ggtgcatggg gagagccagg gtaggaaggg aatgaggaca
      121 cctgtgcagg gggcattggt gggtctgaga cgcacttcct gtcagatctt ctttgagatg
      181 ctaggtaaga agtgtccctt gctgagtgcc ttggagccga ttcacaccgc ctcattaatc
      241 tttgaaacag ctaacagagg cagtgagaac agcaatactg tgtccatctg gagggagagg
      301 aagcagaggc tcacgccggc ggagctcggg ctgggcataa aagtcagggc agagccatct
      361 attgcttaca tttgcttctg ggatccagat ctttcgaagc tagcgctacc ggtcgccacc
      421 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
      481 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
      541 ggcaagctga ccctgaagct gatctgcacc accggcaagc tgcccgtgcc ctggcccacc
      601 ctcgtgacca ccctgggcta cggcgtgcag tgcttcgccc gctaccccga ccacatgaag
      661 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
      721 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
      781 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
      841 aagctggagt acaactacaa cagccacaac gtctatatca ccgccgacaa gcagaagaac
      901 ggcatcaagg ccaacttcaa gatccgccac aacatcgagg acggcggcgt gcagctcgcc
      961 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     1021 tacctgagct accagtccaa gctgagcaaa gaccccaacg agaagcgcga tcacatggtc
     1081 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa
     1141 gtcgacggcg cgccgcggcc gcgaattcga tatcataatc aacctctgga ttacaaaatt
     1201 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     1261 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     1321 tataaatcct ggttagttct tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     1381 tggacagggg ctcggctgtt gggcactgac aattccgtgg ctcgagagat cttcgactgt
     1441 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     1501 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     1561 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     1621 agacaatagc aggcatgaga tctcacgtgc ggaccgagcg gccgcaggaa cccctagtga
     1681 tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg
     1741 tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc
     1801 tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     1861 tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     1921 ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt
     1981 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     2041 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg
     2101 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     2161 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca actctatctc
     2221 gggctattct tttgatttat aagggatttt gccgatttcg gtctattggt taaaaaatga
     2281 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg
     2341 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
     2401 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     2461 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     2521 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     2581 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     2641 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     2701 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     2761 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     2821 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     2881 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     2941 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     3001 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     3061 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     3121 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     3181 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     3241 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     3301 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     3361 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     3421 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     3481 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     3541 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     3601 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     3661 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     3721 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     3781 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     3841 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     3901 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     3961 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     4021 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     4081 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     4141 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     4201 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     4261 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     4321 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     4381 ttgctggcct tttgctcaca tgtcctgcag gcagctgcgc gctcgctcgc tcactgaggc
     4441 cgcccgggcg tcgggcgacc tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag
     4501 agggagtggc caactccatc actaggggtt cctgcggccg cacgcgtaga tgactggagg
     4561 agttgaccag gctgtggctg gggtgctggg agcaggagaa agaggagaag tgggtagcct
     4621 aggagaggaa acagcctgtg ataaaaaaga acagtcagga cctctaggca tccagaggaa
     4681 ggcagctggg ctggcacaca ccagatgtag ctgctgtggg acactgtggg caggaccagg
     4741 tcaaggtttt ggattgggtc ccaggactgc tgggaagcca ttgag
//
