LOCUS       sequence-432552-        5047 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2412m 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 5047)
  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 5047)
  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: AiP1711 - pAAV-AiE2412m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..5047
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     15..34
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   176..305
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      1234..1243
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1240..1242,1243..1245,1246..1956)
                     /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(1285..1306)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1546..1565)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1893..1914
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2049..2069)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2256..2367
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2485..2625
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2485..2614
                     /label=AAV2 ITR
     rep_origin      2700..3155
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2787..2806)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2997..3018
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3172..3191)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3291..3313
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3351..3369)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3437..3541
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3542..3610,3611..4402)
                     /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(3760..3779)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(4573..5047,1..114)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
       61 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
      121 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgtcctgc
      181 aggcagctgc gcgctcgctc gctcactgag gccgcccggg cgtcgggcga cctttggtcg
      241 cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg
      301 ttcctgcggc cgcacgcgtt gtaagagtaa taatcggatc agctccataa caaagaacta
      361 actggtccag aatgtcaata gtgctgaggc tggaaactgg tctcacacac acacacacac
      421 acacacacag agcctaaact ccttcatact gcaaaatgtt tcagaatcat agttggagca
      481 agaaaataga tctgccatcc attatgttat gtgagactct gttgcatgca tggtaggagc
      541 caatatgtta attagttgtc tttttcctaa gtgttcctgc cagtcagtca cctcctgctc
      601 cccaactaac acatattctg aaataagatc tgcctccagt attggcagcg cacaaccata
      661 actctcgaca tttagatgca ggcacttttt gtttcaagga cacaagacgc tgagaaacac
      721 cattctggga cctgaatgat gtcatttcct agaaagcatg ttcaaggtga catcatgctg
      781 catcaggctt gagtagcagg tcctgaggga tggggctacc catgctctcc ctcctaaact
      841 agagtttatg acttgtaaat gggactgtag gtgcaactcc ccagacagga tcttctctcc
      901 gggcaaaggt gataccaaga gatgtgtccc agtatccatc ttggatccca agcagaactc
      961 cagagagcag gaaagctgtt aacaccagat cgagagcact ttccccagac tcctgcggaa
     1021 cagcatgctg tttggctgcc aaaatattag gactaaaagc ggctggcaaa gctgctcatt
     1081 ggggaggtga agctgcacag aggagcctga ggaaaaactt catctttgaa tacggcttcg
     1141 gagctcgggc tgggcataaa agtcagggca gagccatcta ttgcttacat ttgcttctgg
     1201 gatccagatc tttcgaagct agcgctaccg gtcgccacca tggtgagcaa gggcgaggag
     1261 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     1321 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagctg
     1381 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgggctac
     1441 ggcgtgcagt gcttcgcccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     1501 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
     1561 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     1621 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
     1681 agccacaacg tctatatcac cgccgacaag cagaagaacg gcatcaaggc caacttcaag
     1741 atccgccaca acatcgagga cggcggcgtg cagctcgccg accactacca gcagaacacc
     1801 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcta ccagtccaag
     1861 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     1921 gccgggatca ctctcggcat ggacgagctg tacaagtaag tcgacggcgc gccgcggccg
     1981 cgaattcgat atcataatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     2041 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     2101 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttagttctt
     2161 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     2221 ggcactgaca attccgtggc tcgagagatc ttcgactgtg ccttctagtt gccagccatc
     2281 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     2341 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     2401 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgagat
     2461 ctcacgtgcg gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc
     2521 tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg
     2581 cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt
     2641 attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta
     2701 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     2761 tacacttgcc agcgccttag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     2821 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     2881 tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc
     2941 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     3001 actcttgttc caaactggaa caacactcaa ctctatctcg ggctattctt ttgatttata
     3061 agggattttg ccgatttcgg tctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     3121 cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg
     3181 ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg
     3241 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     3301 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat
     3361 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
     3421 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     3481 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     3541 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     3601 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     3661 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     3721 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     3781 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     3841 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     3901 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     3961 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     4021 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     4081 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     4141 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     4201 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     4261 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     4321 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     4381 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     4441 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     4501 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     4561 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     4621 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     4681 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     4741 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     4801 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     4861 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     4921 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     4981 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     5041 gcgcacg
//
