LOCUS       sequence-432550-        5043 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2383m 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 5043)
  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 5043)
  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: AiP1704 - pAAV-AiE2383m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..5043
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     132..153
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(307..326)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     426..448
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(486..504)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        572..676
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(677..745,746..1537)
                     /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(895..914)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1708..2296
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2197..2216
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2358..2487
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      3412..3421
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3418..3420,3421..3423,3424..4134)
                     /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(3463..3484)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3724..3743)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4071..4092
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4227..4247)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    4434..4545
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4663..4803
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4663..4792
                     /label=AAV2 ITR
     rep_origin      join(4878..5043,1..290)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4965..4984)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg
       61 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
      121 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaactcta tctcgggcta
      181 ttcttttgat ttataaggga ttttgccgat ttcggtctat tggttaaaaa atgagctgat
      241 ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt tatggtgcac
      301 tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc
      361 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
      421 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg
      481 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
      541 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
      601 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
      661 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
      721 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
      781 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
      841 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
      901 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
      961 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     1021 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     1081 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     1141 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     1201 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     1261 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     1321 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     1381 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     1441 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     1501 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     1561 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     1621 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     1681 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     1741 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     1801 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     1861 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     1921 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     1981 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     2041 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     2101 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     2161 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     2221 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     2281 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     2341 gccttttgct cacatgtcct gcaggcagct gcgcgctcgc tcgctcactg aggccgcccg
     2401 ggcgtcgggc gacctttggt cgcccggcct cagtgagcga gcgagcgcgc agagagggag
     2461 tggccaactc catcactagg ggttcctgcg gccgcacgcg ttgtaagagt aataatcgca
     2521 ctgggagaag agcagaggaa gccagcttca cccccagtct tcaggtcttg gtggtggcaa
     2581 ggatggagac acttgactga gctgttcccg gctggtagtc aacaatgagg tttaaaggga
     2641 gaggggctga tgagttattg gcaaggggtt caggagaaac acctattgag ctagacctgg
     2701 catgtgactt tctgagaaat cacaggttac agcatttctt agagtgcctg ccttggctgc
     2761 ctctggaaag tgggctgctg gacaacctca gcttactgct gggcctggtg tgtgtgtgtg
     2821 tgtgtgtgtg tgtgtgtgtg tgtgcatgcg cgcgtgtctg ggtatgtgac tgcagcctgc
     2881 attttctccc ctaccagcct tcctcctccc ctccagccac cctgggattg gtgactctca
     2941 gcccctcccc ttgactcccc caaaccctcc cagaaccttt atcagggagc tgggactggt
     3001 ttctgccacc ttccagagcc cctgcactct tgtctctgtc tctgcagcag catcagccag
     3061 ctccctgatc ctgtcagcta tccagctcca caggtaggtt cagggaggac attgcggtag
     3121 cctgaagctg gcaccaaaga tgaggggtgt gagttatgag gggacaaaag ggacagtagc
     3181 ctctgatggg caagcagggc accagaaagg agatcaggaa atgcctggca gggtgggagc
     3241 tgttaatgct tggcatctgg tgccagaatc aagactgatt ctgacaggtc agaaagtttc
     3301 tgtataagtc ttgggtcgga gctcgggctg ggcataaaag tcagggcaga gccatctatt
     3361 gcttacattt gcttctggga tccagatctt tcgaagctag cgctaccggt cgccaccatg
     3421 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
     3481 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
     3541 aagctgaccc tgaagctgat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
     3601 gtgaccaccc tgggctacgg cgtgcagtgc ttcgcccgct accccgacca catgaagcag
     3661 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
     3721 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
     3781 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
     3841 ctggagtaca actacaacag ccacaacgtc tatatcaccg ccgacaagca gaagaacggc
     3901 atcaaggcca acttcaagat ccgccacaac atcgaggacg gcggcgtgca gctcgccgac
     3961 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     4021 ctgagctacc agtccaagct gagcaaagac cccaacgaga agcgcgatca catggtcctg
     4081 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaagtc
     4141 gacggcgcgc cgcggccgcg aattcgatat cataatcaac ctctggatta caaaatttgt
     4201 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     4261 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     4321 aaatcctggt tagttcttgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     4381 acaggggctc ggctgttggg cactgacaat tccgtggctc gagagatctt cgactgtgcc
     4441 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     4501 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     4561 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     4621 caatagcagg catgagatct cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg
     4681 agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg
     4741 cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc
     4801 aggggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac
     4861 gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     4921 tacgcgcagc gtgaccgcta cacttgccag cgccttagcg cccgctcctt tcgctttctt
     4981 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     5041 ttt
//
