LOCUS       sequence-433030-        6087 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2193m 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 6087)
  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 6087)
  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: AiP1514 - pAAV-AiE2193m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..6087
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     235..256
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(391..411)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    598..709
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   827..967
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   827..956
                     /label=AAV2 ITR
     rep_origin      1042..1497
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1129..1148)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1339..1360
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1514..1533)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1633..1655
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1693..1711)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1779..1883
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1884..1952,1953..2744)
                     /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(2102..2121)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2915..3503
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3404..3423
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   3565..3694
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     CDS             complement(4608..4619)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     regulatory      5663..5672
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(5669..5671,5672..5674,5675..6087,1..298)
                     /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(5714..5735)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5975..5994)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
ORIGIN
        1 caagctggag tacaactaca acagccacaa cgtctatatc accgccgaca agcagaagaa
       61 cggcatcaag gccaacttca agatccgcca caacatcgag gacggcggcg tgcagctcgc
      121 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
      181 ctacctgagc taccagtcca agctgagcaa agaccccaac gagaagcgcg atcacatggt
      241 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta
      301 agtcgacggc gcgccgcggc cgcgaattcg atatcataat caacctctgg attacaaaat
      361 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
      421 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
      481 gtataaatcc tggttagttc ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
      541 ctggacaggg gctcggctgt tgggcactga caattccgtg gctcgagaga tcttcgactg
      601 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
      661 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
      721 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
      781 aagacaatag caggcatgag atctcacgtg cggaccgagc ggccgcagga acccctagtg
      841 atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag
      901 gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc
      961 ctgcaggggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     1021 atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     1081 tggttacgcg cagcgtgacc gctacacttg ccagcgcctt agcgcccgct cctttcgctt
     1141 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     1201 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg
     1261 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     1321 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aactctatct
     1381 cgggctattc ttttgattta taagggattt tgccgatttc ggtctattgg ttaaaaaatg
     1441 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat
     1501 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc
     1561 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     1621 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     1681 cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
     1741 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     1801 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     1861 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     1921 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     1981 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     2041 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     2101 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     2161 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     2221 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     2281 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     2341 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     2401 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     2461 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     2521 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     2581 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     2641 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     2701 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     2761 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     2821 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     2881 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     2941 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     3001 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     3061 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     3121 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     3181 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     3241 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     3301 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     3361 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     3421 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     3481 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     3541 tttgctggcc ttttgctcac atgtcctgca ggcagctgcg cgctcgctcg ctcactgagg
     3601 ccgcccgggc gtcgggcgac ctttggtcgc ccggcctcag tgagcgagcg agcgcgcaga
     3661 gagggagtgg ccaactccat cactaggggt tcctgcggcc gcacgcgttg taagagtaat
     3721 aatcgccggc gccaattgga tatagttttc acctctcagt tattgttagt agtgctaaga
     3781 caaacttgga agtacaatgc ttccccgaca ctgatttcat tccctttgta gatattccaa
     3841 gtgacagact actgcatcat atggtattac tatttttaac attaaggggc catcacaacc
     3901 tctttctttt atataatagc tgtaaaatgt acattgctgc taatactgta caagaactct
     3961 cttttgtctt ccctgtcact tgttctccct ttgttaatac ccatttttta tgggtggcat
     4021 taaaacgcag tgtgattttg agctacatgt tcctaatgat tgggaatgtt gaaagatttt
     4081 tttttttaac atacaacttt tggccatccc catgttcttt tttgagaaat gtttgtttaa
     4141 ggtctgtcac ttagttttta ggtgagctaa gtgtctgtta gatattgagt tgtataaaat
     4201 ctttttgcag tctgggcatt aactttttaa atttaattca atttatttta gttattcact
     4261 tcacatcctg ctcactgccc tcctacaagc cacctcctcc cacaatactt cccccttatg
     4321 agtattcccc ctaccaccct ggcacctcaa gtctctgcaa ggcaaggcta ggcaacctag
     4381 ctattaggtc atatcccagg caagtgcttt tggatagccc ttgttccagt tgttcaggac
     4441 ccacatgaag atcaagctaa acatctgcta catatgagcc gggagttcta ggtctagccc
     4501 tgtatgttct ttgattgatg gttcagactg ggagcgccct gagagtccag gttaatggac
     4561 tcacttgatc ttcctgtggc gtccctatcc ctttcggggt ccacaatcct tccttctatt
     4621 gttccataag agaccccaag ttccatccac tgtttggctg tgggtgtctg tatatctctg
     4681 agtcagctgc tgtgtgaagc ctctcagagg aaaatatgct actttcagta agcatgacag
     4741 agtatcatta gtagtgctag ggattagtgc ttgcacatgg gatgggtttc aagttggaac
     4801 agttattggt tgaccattcc ctcagtctct gttcctctcc atgtgcctgt cttacttgta
     4861 gacaggattt tgagttgaaa gttttgtggg tgggttggca tctctatcac tccactgggg
     4921 ttcctgtctg tcaacaggag gtagcctata cagcttccat atccccaatg ccatgagtca
     4981 tagctaaggt cacccccact cgtcctggag atgcccccac aaccttaccc ccatcagctg
     5041 cacatttcca ttcatttttt caataacttc ttatatgtct agtatggaat ggagtttttt
     5101 cctttttagt ctcataccta gagtttcttt agtgtgaaat ccagcacttc tcaaattagt
     5161 attcactgcc aagctttaaa aagggggcaa tgtaaacctt gttctgtcta taaataacag
     5221 gaaaagtgac aaactgactt aaagagtaag atttggtgtt tcttcttaga cagaaggtag
     5281 agtggcccca ctttggccaa tttggtagct tcacagtgaa atcaaagaat tatgacctgt
     5341 ccttctcttt catatttctt gtattatcta cattcaagtc ttgaacaact cttgtttacc
     5401 atgcttccag gtaaccttgc taagggggga aaaatcacct gtaaaaggga attctttaat
     5461 taagaggaga aaaaatttcc ctcagagatc tcatctctgg tcatactgcc cagacatgag
     5521 tggcactgcc ccctccccca tcaaccatga ccagagtgaa tcgccggcgg agctcgggct
     5581 gggcataaaa gtcagggcag agccatctat tgcttacatt tgcttctggg atccagatct
     5641 ttcgaagcta gcgctaccgg tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg
     5701 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc
     5761 cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagctga tctgcaccac
     5821 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgggctacg gcgtgcagtg
     5881 cttcgcccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
     5941 aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc
     6001 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt
     6061 caaggaggac ggcaacatcc tggggca
//
