LOCUS       sequence-466243-        5490 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2083m 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 5490)
  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 5490)
  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: AiP1360 - pAAV-AiE2083m-minBG-SYFP2-4X2C-WPRE3...
FEATURES             Location/Qualifiers
     source          1..5490
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     164..183
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   325..454
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      1606..1615
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1612..1614,1615..1617,1618..2328)
                     /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(1657..1678)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1918..1937)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2265..2286
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2641..2661)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2848..2959
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3077..3217
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3077..3206
                     /label=AAV2 ITR
     rep_origin      3292..3747
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3379..3398)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3589..3610
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3764..3783)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3883..3905
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3943..3961)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4029..4133
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4134..4202,4203..4994)
                     /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(4352..4371)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(5165..5490,1..263)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
       61 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
      121 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
      181 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
      241 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
      301 tttgctggcc ttttgctcac atgtcctgca ggcagctgcg cgctcgctcg ctcactgagg
      361 ccgcccgggc gtcgggcgac ctttggtcgc ccggcctcag tgagcgagcg agcgcgcaga
      421 gagggagtgg ccaactccat cactaggggt tcctgcggcc gcacgcgtct ggctgtcctg
      481 gaactcactc tgtagaccag gctggcctca aactcagaaa tctgcctgcc tctgcctccc
      541 aagtgctggg attaaaggca tgtgccacca cacctggcct agaaataatg acttttaaaa
      601 ttatctaaag gttaccagat gaaattatca acctcagctc tgtggtataa tttaaatcta
      661 gttctgattc agtaagagtg ttattttcag taccctcagt agcagaatcc tattgcctct
      721 tatatctaag caggagaggc aattcagaaa caaagaccca aggacaggag ccctattgca
      781 tctactctca ataattaagg cctttgatga ctaaaataat ttagataata agattggttt
      841 gggtaaataa atatcaaatt agggtttatg gcagtcacag gtgtttggca ggtctggcct
      901 ataatcacag agaaaatgaa tgcattttat ttagcataat tgggaaaagg aatgaagctt
      961 atatgcaaat tgtgtaccac ttttgttaaa gatagctggt cagactcatg atgataatcc
     1021 aaacctttat cccagtgaaa attttcaaca tatctatttt ctgtggaatt ttaccacatc
     1081 tgactgcatt ctcccagtct tctgagatgg atttcagtgt ccttggtcac catctagcct
     1141 ttctattttg ttgacaggat gcctgagttt agctgtttct gtttatttgt ttgtttttaa
     1201 gcttgatttc tcctactcgt gtctatagct ggaatcggag gttacatgag atttctcagc
     1261 atcagcatcc agcactgcca caaaggggat gtgggagatg agaagggaaa acaagacagt
     1321 gaagagaata aaaatgaaat cctggaggca agataattaa gagaccagaa aatagagttg
     1381 aatttccatt aggaacattt acaagaatgt gttcacaggc acacacacag gaatccccaa
     1441 ctgctagctt tggaatgccc atagatgcca ctgctactgg actctgtgat cagcgctctt
     1501 gactaggact acgagctcgg gctgggcata aaagtcaggg cagagccatc tattgcttac
     1561 atttgcttct gggatccaga tctttcgaag ctagcgctac cggtcgccac catggtgagc
     1621 aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta
     1681 aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg
     1741 accctgaagc tgatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc
     1801 accctgggct acggcgtgca gtgcttcgcc cgctaccccg accacatgaa gcagcacgac
     1861 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac
     1921 gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
     1981 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag
     2041 tacaactaca acagccacaa cgtctatatc accgccgaca agcagaagaa cggcatcaag
     2101 gccaacttca agatccgcca caacatcgag gacggcggcg tgcagctcgc cgaccactac
     2161 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc
     2221 taccagtcca agctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag
     2281 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta agtcgacggc
     2341 gcgccgcggc cttaaagaga ccggttcact gtgacagtaa aagagaccgg ttcactgtga
     2401 gaatgaaaga gaccggttca ctgtgatcgg aaaagagacc ggttcactgt gagcggcctt
     2461 gaaacccagc agacaatgta gctcagtaga aacccagcag acaatgtagc tgaatggaaa
     2521 cccagcagac aatgtagctt cggagaaacc cagcagacaa tgtagctgtc gacgaattcg
     2581 atatcataat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     2641 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     2701 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttagttc ttgccacggc
     2761 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     2821 caattccgtg gctcgagaga tcttcgactg tgccttctag ttgccagcca tctgttgttt
     2881 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     2941 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     3001 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgag atctcacgtg
     3061 cggaccgagc ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct
     3121 cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg
     3181 gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc
     3241 cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct
     3301 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     3361 ccagcgcctt agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     3421 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     3481 ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct
     3541 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     3601 tccaaactgg aacaacactc aactctatct cgggctattc ttttgattta taagggattt
     3661 tgccgatttc ggtctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     3721 ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg
     3781 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     3841 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     3901 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat
     3961 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     4021 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     4081 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     4141 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     4201 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     4261 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     4321 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     4381 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     4441 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     4501 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     4561 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     4621 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     4681 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     4741 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     4801 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     4861 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     4921 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     4981 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     5041 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     5101 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     5161 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     5221 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     5281 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     5341 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     5401 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     5461 ataaggcgca gcggtcgggc tgaacggggg
//
