LOCUS       sequence-432565-        4925 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2487m 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 4925)
  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 4925)
  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: AiP1834 - pAAV-AiE2487m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4925
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     91..112
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(266..285)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     385..407
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(445..463)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        531..635
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(636..704,705..1496)
                     /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(854..873)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1667..2255
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2156..2175
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2317..2446
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      3253..3262
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3259..3261,3262..3264,3265..3975)
                     /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(3304..3325)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3565..3584)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3912..3933
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4068..4088)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    4275..4386
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4504..4644
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4504..4633
                     /label=AAV2 ITR
     rep_origin      join(4719..4925,1..249)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4806..4825)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
       61 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
      121 tcaactctat ctcgggctat tcttttgatt tataagggat tttgccgatt tcggtctatt
      181 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt
      241 ttacaatttt atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc
      301 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
      361 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
      421 caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca
      481 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
      541 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
      601 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
      661 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
      721 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
      781 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
      841 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
      901 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
      961 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     1021 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     1081 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     1141 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     1201 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     1261 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     1321 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     1381 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     1441 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     1501 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     1561 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     1621 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     1681 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     1741 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     1801 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     1861 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     1921 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     1981 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     2041 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     2101 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     2161 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     2221 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     2281 ggttcctggc cttttgctgg ccttttgctc acatgtcctg caggcagctg cgcgctcgct
     2341 cgctcactga ggccgcccgg gcgtcgggcg acctttggtc gcccggcctc agtgagcgag
     2401 cgagcgcgca gagagggagt ggccaactcc atcactaggg gttcctgcgg ccgcacgcgt
     2461 tgtaagagta ataatcgatg caattggaga tagatcaagc catctaagcc agtcttgaaa
     2521 agatcagttt ctatgatttt ctaagaaaag atttctatga tttgtggttc ctaaatttta
     2581 tataaaacca catgtgtata tatgaaatta agtcagaagt gaagttatcc agggggagaa
     2641 cagggacaaa tgtgagggag tgtggaagga agggacagca ggacatacac tagagatgca
     2701 gtaaacacct gtctgaaatg ttaaaaaata aaatacaaaa gaaagagtgc tagagaaagc
     2761 gaatgagatc aaagcatatt atccatccct gtgctctctt ttgatctggg ttgctgtgac
     2821 aaaatgccta gcaaacagca cagcaggacg ggaaagggct tggctttcaa tcgcaggcta
     2881 ccatctgtca tgtcagggaa gacaaggcag aaacacatgc aactaatcac atgactccca
     2941 cagtcaagag cggagactaa agaaagcccc cacactgcct atagctggct tagcacccag
     3001 ccggcttctt ctactctcat gtaatccagg ccttgcccag agaatgaatg gcaccaccaa
     3061 caatgaaata cgtcttccta tcaagctaat aatcaagaca tgcccacggg tcaatgtgct
     3121 ctacttagtt ctccctgtta ggctctttcc ccaatgacgg agctcgggct gggcataaaa
     3181 gtcagggcag agccatctat tgcttacatt tgcttctggg atccagatct ttcgaagcta
     3241 gcgctaccgg tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
     3301 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc
     3361 gagggcgatg ccacctacgg caagctgacc ctgaagctga tctgcaccac cggcaagctg
     3421 cccgtgccct ggcccaccct cgtgaccacc ctgggctacg gcgtgcagtg cttcgcccgc
     3481 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
     3541 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
     3601 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
     3661 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcacc
     3721 gccgacaagc agaagaacgg catcaaggcc aacttcaaga tccgccacaa catcgaggac
     3781 ggcggcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     3841 ctgctgcccg acaaccacta cctgagctac cagtccaagc tgagcaaaga ccccaacgag
     3901 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
     3961 gacgagctgt acaagtaagt cgacggcgcg ccgcggccgc gaattcgata tcataatcaa
     4021 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     4081 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     4141 ttcattttct cctccttgta taaatcctgg ttagttcttg ccacggcgga actcatcgcc
     4201 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggct
     4261 cgagagatct tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     4321 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     4381 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     4441 caagggggag gattgggaag acaatagcag gcatgagatc tcacgtgcgg accgagcggc
     4501 cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg
     4561 aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg
     4621 agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt acgcatctgt
     4681 gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt
     4741 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccttagc
     4801 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     4861 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     4921 caaaa
//
