LOCUS       sequence-432540-        5149 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2167m 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 5149)
  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 5149)
  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: AiP1438 - pAAV-AiE2167m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..5149
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     189..210
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(364..383)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     483..505
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(543..561)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        629..733
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(734..802,803..1594)
                     /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(952..971)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1765..2353
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2254..2273
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2415..2544
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      3575..3584
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3581..3583,3584..3586,3587..4297)
                     /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(3626..3647)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3887..3906)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4234..4255
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4390..4410)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    4597..4708
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4826..4966
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4826..4955
                     /label=AAV2 ITR
     rep_origin      join(5041..5149,1..347)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5128..5147)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
       61 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc
      121 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
      181 ctttaatagt ggactcttgt tccaaactgg aacaacactc aactctatct cgggctattc
      241 ttttgattta taagggattt tgccgatttc ggtctattgg ttaaaaaatg agctgattta
      301 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct
      361 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc
      421 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
      481 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa
      541 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
      601 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
      661 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
      721 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
      781 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
      841 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
      901 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
      961 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     1021 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     1081 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     1141 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     1201 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     1261 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     1321 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     1381 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     1441 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     1501 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     1561 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     1621 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     1681 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     1741 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     1801 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     1861 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     1921 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     1981 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     2041 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     2101 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     2161 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     2221 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     2281 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     2341 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     2401 ttttgctcac atgtcctgca ggcagctgcg cgctcgctcg ctcactgagg ccgcccgggc
     2461 gtcgggcgac ctttggtcgc ccggcctcag tgagcgagcg agcgcgcaga gagggagtgg
     2521 ccaactccat cactaggggt tcctgcggcc gcacgcgttg taagagtaat aatcgccggc
     2581 gagtaactgt gcaagggtga aaaaaagtcc tagagtgagc acagggctgg catgcagggt
     2641 ctgatgctag agtgcatctc ctaggctctt agcctctttt tgctcagtct agctttcttt
     2701 acactctact ggagctcaaa gtttatcttt taatgataaa gtaaaaatgc tgtagccata
     2761 tctgttggag ccagttgtga ggcagggaat gtagtccctg gtttttcttt gttttttgtt
     2821 tttgtttgtt tgtttgtttg tttgttttgt tttgagacag ggtttctctg tgtagctcta
     2881 ggtgtcctgg aacttgctct atagtcctag gctggcttct tactcagaga tctcccttct
     2941 ctgcctccca agtgctgtga tcaaaggcat gccccaccac tgcacagttc cagtctctga
     3001 tttaatgtgt agacatgtat tgctctgtgt ttagaattaa gcaacttaga cccaaagaaa
     3061 gcaggatgag ttttaaaggg gctgtcatca ttctgactca caggtgatgt ggcagtcgaa
     3121 caagacagca tctgcccttt ctaagacagg accagggaaa tcagaacctt tgcctgatga
     3181 ctgcttagtg tgaagcagct agacacagcc ttgatttgtt taatgcatct ttctctgcca
     3241 tctcattagt gacacgagct gctaattatg tatccagaag ggctaatttc ccctgttcct
     3301 ctgtcacttt ttgagaggat tcaggcctgg gtgtagaagc aggagaatgg tcattaagtc
     3361 catgatgctt cagggatgtg gttacacact tacaaatggc aagtcaaaag gataatcctt
     3421 aagagccttg atttcgagag ctttagttga gtgtactgca cttgacaact gagcgccggc
     3481 ggagctcggg ctgggcataa aagtcagggc agagccatct attgcttaca tttgcttctg
     3541 ggatccagat ctttcgaagc tagcgctacc ggtcgccacc atggtgagca agggcgagga
     3601 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
     3661 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagct
     3721 gatctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgggcta
     3781 cggcgtgcag tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     3841 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
     3901 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     3961 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
     4021 cagccacaac gtctatatca ccgccgacaa gcagaagaac ggcatcaagg ccaacttcaa
     4081 gatccgccac aacatcgagg acggcggcgt gcagctcgcc gaccactacc agcagaacac
     4141 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagct accagtccaa
     4201 gctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
     4261 cgccgggatc actctcggca tggacgagct gtacaagtaa gtcgacggcg cgccgcggcc
     4321 gcgaattcga tatcataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     4381 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     4441 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttagttct
     4501 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     4561 gggcactgac aattccgtgg ctcgagagat cttcgactgt gccttctagt tgccagccat
     4621 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     4681 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     4741 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgaga
     4801 tctcacgtgc ggaccgagcg gccgcaggaa cccctagtga tggagttggc cactccctct
     4861 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt
     4921 gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg
     4981 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt
     5041 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     5101 ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttc
//
