LOCUS       sequence-432537-        4843 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2112m 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 4843)
  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 4843)
  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: AiP1420 - pAAV-AiE2112m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4843
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     172..191
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   353..493
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   364..493
                     /label=AAV2 ITR
     polyA_signal    611..722
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     909..929
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(1022..1732,1733..1735,1736..1738))
                     /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(1064..1085)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1413..1432
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1672..1693
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1735..1744
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   2469..2598
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(2660..3248)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2740..2759)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3419..4210,4211..4279))
                     /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     4042..4061
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4280..4384)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4452..4470
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4508..4530)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4630..4649
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(join(4666..4843,1..278))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4803..4824)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
       61 caaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      121 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      181 agcgaaagga gcgggcgcta aggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      241 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtac tatggttgct ttgacgtatg
      301 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg cccctgcagg
      361 cagctgcgcg ctcgctcgct cactgaggcc gcccgggcaa agcccgggcg tcgggcgacc
      421 tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc
      481 actaggggtt cctgcggccg ctcggtccgc acgtgagatc tcatgcctgc tattgtcttc
      541 ccaatcctcc cccttgctgt cctgccccac cccacccccc agaatagaat gacacctact
      601 cagacaatgc gatgcaattt cctcatttta ttaggaaagg acagtgggag tggcaccttc
      661 cagggtcaag gaaggcacgg gggaggggca aacaacagat ggctggcaac tagaaggcac
      721 agtcgaagat ctctcgagcc acggaattgt cagtgcccaa cagccgagcc cctgtccagc
      781 agcgggcaag gcaggcggcg atgagttccg ccgtggcaag aactaaccag gatttataca
      841 aggaggagaa aatgaaagcc atacgggaag caatagcatg atacaaaggc attaaagcag
      901 cgtatccaca tagcgtaaaa ggagcaacat agttaagaat accagtcaat ctttcacaaa
      961 ttttgtaatc cagaggttga ttatgatatc gaattcgcgg ccgcggcgcg ccgtcgactt
     1021 acttgtacag ctcgtccatg ccgagagtga tcccggcggc ggtcacgaac tccagcagga
     1081 ccatgtgatc gcgcttctcg ttggggtctt tgctcagctt ggactggtag ctcaggtagt
     1141 ggttgtcggg cagcagcacg gggccgtcgc cgatgggggt gttctgctgg tagtggtcgg
     1201 cgagctgcac gccgccgtcc tcgatgttgt ggcggatctt gaagttggcc ttgatgccgt
     1261 tcttctgctt gtcggcggtg atatagacgt tgtggctgtt gtagttgtac tccagcttgt
     1321 gccccaggat gttgccgtcc tccttgaagt cgatgccctt cagctcgatg cggttcacca
     1381 gggtgtcgcc ctcgaacttc acctcggcgc gggtcttgta gttgccgtcg tccttgaaga
     1441 agatggtgcg ctcctggacg tagccttcgg gcatggcgga cttgaagaag tcgtgctgct
     1501 tcatgtggtc ggggtagcgg gcgaagcact gcacgccgta gcccagggtg gtcacgaggg
     1561 tgggccaggg cacgggcagc ttgccggtgg tgcagatcag cttcagggtc agcttgccgt
     1621 aggtggcatc gccctcgccc tcgccggaca cgctgaactt gtggccgttt acgtcgccgt
     1681 ccagctcgac caggatgggc accaccccgg tgaacagctc ctcgcccttg ctcaccatgg
     1741 tggcgaccgg tagcgctagc ttcgaaagat ctggatccca gaagcaaatg taagcaatag
     1801 atggctctgc cctgactttt atgcccagcc cgagctccgc cggcggactc tttcaagcca
     1861 actagctaaa tggggtgata gaattctgca ttttcacaga agcagctaga gtttgaggtc
     1921 tgttgcctta cgtagaatga gaaggtgctg cacaagtcaa ggcactgcta ctctgagctt
     1981 ttaggttatg atacatccat aaggttcctt cctcagcaag gatagaagac agatgttaac
     2041 ctgaaagcca gccataagga gctttggcac ttgtgtggca gataagctgt ttccagactg
     2101 cgggacacag ctagtgccac gttgggaaaa ggtcaacaga agctgacttt ccattccagc
     2161 tttatgcttt ctagaagcct gagaagctga agaacagcct gttcctgcat agttacaatt
     2221 cttctgggct ctttcttaag tgttctttgg ggttagcaga aggagtgggc gcacacacac
     2281 acgcgcacgc gtgcacacac acacacacgt tcttttccaa gacagttgcc ctttggggaa
     2341 ctggaagcat acacttagat ataaacagga gcagggttct aaagtttact gagcccggag
     2401 ggctgctcct ctcttggagg gctgtgtcct gcgccggcga ttattactct tacaacgcgt
     2461 gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct
     2521 cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc ggcctcagtg agcgagcgag
     2581 cgcgcagctg cctgcaggac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     2641 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     2701 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     2761 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     2821 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     2881 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     2941 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     3001 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     3061 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     3121 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     3181 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     3241 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     3301 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     3361 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     3421 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     3481 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     3541 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     3601 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     3661 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     3721 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     3781 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     3841 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     3901 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     3961 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     4021 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     4081 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     4141 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     4201 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     4261 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     4321 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     4381 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
     4441 taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg
     4501 gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg
     4561 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc
     4621 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccataaaatt gtaaacgtta
     4681 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaataga
     4741 ccgaaatcgg caaaatccct tataaatcaa aagaatagcc cgagatagag ttgagtgttg
     4801 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctc
//
