LOCUS       sequence-466296-        4944 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2396m 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 4944)
  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 4944)
  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: AiP1727 - pAAV-AiE2396m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4944
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   732..861
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(923..1511)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1003..1022)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1682..2473,2474..2542))
                     /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     2305..2324
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2543..2647)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2715..2733
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2771..2793)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2893..2912
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2929..3384)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3066..3087)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3278..3297
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   3459..3599
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3470..3599
                     /label=AAV2 ITR
     polyA_signal    3717..3828
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     4015..4035
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(4128..4838,4839..4841,4842..4844))
                     /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(4170..4191)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4519..4538
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4778..4799
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4841..4850
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 ggtttttttt ttgctagctt atatgacact catttttttt acccgttgaa taattcatgt
       61 accactgacc agcacaataa gatatttcac tttgagcgtg catctacatt tcttaactta
      121 gctctcaatt ctaattattg cagccaattt ctgttccttg atgagaagag gaatcctaca
      181 tagacctgaa caacaggtaa tgagcttcat taaataggcc agctgagcga ggcttcccta
      241 gacttacaca agcgttcaat ttccttcctg agtcagaact tgagcaaact gttaagcagg
      301 gagaagatta tccacagact tggcagtcgg aggtggtaat actctcctgt cttcactctt
      361 cttcacagtc ctgcctggat gccagaaaag ggagaaacag gcttttcccc ctgcttcatc
      421 ctcacacatg aacacgagtg tctgtgtgtg tgcatgcgtg tacacacaca cacacacaca
      481 cacacacaca cacacacaca caccactctt ctccaaactg ttgttctcag tgagtttgga
      541 gaaagaggaa gagtgggttg ggctatattc cctgaggtcc acatccatgc tttccctgaa
      601 tcttgggtgg gaccgtcctg gtcctcatgg ttcaagaaat ggggagatta tcagagtgct
      661 tctcagagac gcttaagaga gagatagaac ctgctgagct cgattattac tcttacaacg
      721 cgtgcggccg caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc
      781 gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggcggcctca gtgagcgagc
      841 gagcgcgcag ctgcctgcag gacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
      901 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
      961 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     1021 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     1081 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     1141 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     1201 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     1261 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     1321 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     1381 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     1441 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     1501 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     1561 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     1621 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     1681 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     1741 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     1801 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     1861 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     1921 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     1981 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     2041 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     2101 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     2161 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     2221 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     2281 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     2341 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     2401 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     2461 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     2521 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     2581 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     2641 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     2701 cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg
     2761 acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg
     2821 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct
     2881 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccataaa attgtaaacg
     2941 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     3001 agaccgaaat cggcaaaatc ccttataaat caaaagaata gcccgagata gagttgagtg
     3061 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     3121 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcacccaaa tcaagttttt
     3181 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     3241 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     3301 gcgctaaggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     3361 ttaatgcgcc gctacagggc gcgtactatg gttgctttga cgtatgcggt gtgaaatacc
     3421 gcacagatgc gtaaggagaa aataccgcat caggcgcccc tgcaggcagc tgcgcgctcg
     3481 ctcgctcact gaggccgccc gggcaaagcc cgggcgtcgg gcgacctttg gtcgcccggc
     3541 ctcagtgagc gagcgagcgc gcagagaggg agtggccaac tccatcacta ggggttcctg
     3601 cggccgctcg gtccgcacgt gagatctcat gcctgctatt gtcttcccaa tcctccccct
     3661 tgctgtcctg ccccacccca ccccccagaa tagaatgaca cctactcaga caatgcgatg
     3721 caatttcctc attttattag gaaaggacag tgggagtggc accttccagg gtcaaggaag
     3781 gcacggggga ggggcaaaca acagatggct ggcaactaga aggcacagtc gaagatctct
     3841 cgagccacgg aattgtcagt gcccaacagc cgagcccctg tccagcagcg ggcaaggcag
     3901 gcggcgatga gttccgccgt ggcaagaact aaccaggatt tatacaagga ggagaaaatg
     3961 aaagccatac gggaagcaat agcatgatac aaaggcatta aagcagcgta tccacatagc
     4021 gtaaaaggag caacatagtt aagaatacca gtcaatcttt cacaaatttt gtaatccaga
     4081 ggttgattat gatatcgaat tcgcggccgc ggcgcgccgt cgacttactt gtacagctcg
     4141 tccatgccga gagtgatccc ggcggcggtc acgaactcca gcaggaccat gtgatcgcgc
     4201 ttctcgttgg ggtctttgct cagcttggac tggtagctca ggtagtggtt gtcgggcagc
     4261 agcacggggc cgtcgccgat gggggtgttc tgctggtagt ggtcggcgag ctgcacgccg
     4321 ccgtcctcga tgttgtggcg gatcttgaag ttggccttga tgccgttctt ctgcttgtcg
     4381 gcggtgatat agacgttgtg gctgttgtag ttgtactcca gcttgtgccc caggatgttg
     4441 ccgtcctcct tgaagtcgat gcccttcagc tcgatgcggt tcaccagggt gtcgccctcg
     4501 aacttcacct cggcgcgggt cttgtagttg ccgtcgtcct tgaagaagat ggtgcgctcc
     4561 tggacgtagc cttcgggcat ggcggacttg aagaagtcgt gctgcttcat gtggtcgggg
     4621 tagcgggcga agcactgcac gccgtagccc agggtggtca cgagggtggg ccagggcacg
     4681 ggcagcttgc cggtggtgca gatcagcttc agggtcagct tgccgtaggt ggcatcgccc
     4741 tcgccctcgc cggacacgct gaacttgtgg ccgtttacgt cgccgtccag ctcgaccagg
     4801 atgggcacca ccccggtgaa cagctcctcg cccttgctca ccatggtggc gaccggtagc
     4861 gctagcttcg aaagatctgg atcccagaag caaatgtaag caatagatgg ctctgccctg
     4921 acttttatgc ccagcccgag ctcc
//
