LOCUS       sequence-433182-        4903 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2357m 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 4903)
  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 4903)
  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: AiP1930 - pAAV-AiE2357m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4903
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      915..924
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(921..923,924..926,927..1637)
                     /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(966..987)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1227..1246)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1574..1595
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1730..1750)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1937..2048
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2166..2306
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2166..2295
                     /label=AAV2 ITR
     rep_origin      2381..2836
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2468..2487)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2678..2699
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2853..2872)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2972..2994
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3032..3050)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3118..3222
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3223..3291,3292..4083)
                     /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(3441..3460)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4254..4842
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4743..4762
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcaa cctaaaatct ctgtttctcc agatgccatc tctaatgaga
      181 tttccaaggc atgactatgt ttcctaatgc ttgatgttat tgtcgaaaat tggcctgtgt
      241 agttctcaac atctttattt tcagaatgat acagaaagga ccaagtgcct ggttaaaagc
      301 ttagaattaa gcctctgctc acctctgact tgctctgctg gtgtgaatct cagcttctct
      361 aggacttcat ttcttcaacc ataaagctca gagactggat gaggcagcac acagagtctc
      421 tccctgtgct aagattctct ggtttcattt gctggttgtt ttctccccaa acatctggcc
      481 tgggagacag ttggggctgc cagtgtattc tttagtcatg cttagatcaa aaaattatgt
      541 ggcagcccaa tttaaagcat gctctttttt ttttttttta aaaggagcac tgttgcagac
      601 ctaaaaaaat aatgtaggat tcccatttac ctttcataaa gaaagggcga attgttctcc
      661 aaaggctcgg gttgttgtta aatatgatgc cgttctcatg catgccgatg cactgcagcc
      721 cacgcttgct gccgaatcgg gagatgtagt gactgtgctt catgacatgg aacatgcttg
      781 aggacctgtc aagaaagcga gccgtactgt caatccgacc ggagctcggg ctgggcataa
      841 aagtcagggc agagccatct attgcttaca tttgcttctg ggatccagat ctttcgaagc
      901 tagcgctacc ggtcgccacc atggtgagca agggcgagga gctgttcacc ggggtggtgc
      961 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
     1021 gcgagggcga tgccacctac ggcaagctga ccctgaagct gatctgcacc accggcaagc
     1081 tgcccgtgcc ctggcccacc ctcgtgacca ccctgggcta cggcgtgcag tgcttcgccc
     1141 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     1201 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     1261 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     1321 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
     1381 ccgccgacaa gcagaagaac ggcatcaagg ccaacttcaa gatccgccac aacatcgagg
     1441 acggcggcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     1501 tgctgctgcc cgacaaccac tacctgagct accagtccaa gctgagcaaa gaccccaacg
     1561 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
     1621 tggacgagct gtacaagtaa gtcgacggcg cgccgcggcc gcgaattcga tatcataatc
     1681 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
     1741 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
     1801 ctttcatttt ctcctccttg tataaatcct ggttagttct tgccacggcg gaactcatcg
     1861 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg
     1921 ctcgagagat cttcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     1981 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     2041 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     2101 agcaaggggg aggattggga agacaatagc aggcatgaga tctcacgtgc ggaccgagcg
     2161 gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac
     2221 tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag
     2281 cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc ttacgcatct
     2341 gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca
     2401 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta
     2461 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     2521 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     2581 cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     2641 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     2701 acaacactca actctatctc gggctattct tttgatttat aagggatttt gccgatttcg
     2761 gtctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     2821 ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     2881 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     2941 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     3001 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt
     3061 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     3121 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     3181 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     3241 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     3301 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     3361 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     3421 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     3481 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     3541 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     3601 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     3661 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     3721 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     3781 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     3841 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     3901 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     3961 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     4021 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     4081 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     4141 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     4201 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     4261 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     4321 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     4381 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     4441 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     4501 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     4561 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     4621 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     4681 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     4741 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     4801 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     4861 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgt
//
