LOCUS       sequence-438939-        4876 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2545m 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 4876)
  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 4876)
  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: AiP1966 - pAAV-AiE2545m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4876
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   33..173
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   44..173
                     /label=AAV2 ITR
     polyA_signal    291..402
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     589..609
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(702..1412,1413..1415,1416..1418))
                     /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(744..765)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1093..1112
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1352..1373
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1415..1424
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   2182..2311
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(2373..2961)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2453..2472)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3132..3923,3924..3992))
                     /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     3755..3774
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3993..4097)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4165..4183
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4221..4243)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4343..4362
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(4379..4834)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4516..4537)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4728..4747
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 atgcgtaagg agaaaatacc gcatcaggcg cccctgcagg cagctgcgcg ctcgctcgct
       61 cactgaggcc gcccgggcaa agcccgggcg tcgggcgacc tttggtcgcc cggcctcagt
      121 gagcgagcga gcgcgcagag agggagtggc caactccatc actaggggtt cctgcggccg
      181 ctcggtccgc acgtgagatc tcatgcctgc tattgtcttc ccaatcctcc cccttgctgt
      241 cctgccccac cccacccccc agaatagaat gacacctact cagacaatgc gatgcaattt
      301 cctcatttta ttaggaaagg acagtgggag tggcaccttc cagggtcaag gaaggcacgg
      361 gggaggggca aacaacagat ggctggcaac tagaaggcac agtcgaagat ctctcgagcc
      421 acggaattgt cagtgcccaa cagccgagcc cctgtccagc agcgggcaag gcaggcggcg
      481 atgagttccg ccgtggcaag aactaaccag gatttataca aggaggagaa aatgaaagcc
      541 atacgggaag caatagcatg atacaaaggc attaaagcag cgtatccaca tagcgtaaaa
      601 ggagcaacat agttaagaat accagtcaat ctttcacaaa ttttgtaatc cagaggttga
      661 ttatgatatc gaattcgcgg ccgcggcgcg ccgtcgactt acttgtacag ctcgtccatg
      721 ccgagagtga tcccggcggc ggtcacgaac tccagcagga ccatgtgatc gcgcttctcg
      781 ttggggtctt tgctcagctt ggactggtag ctcaggtagt ggttgtcggg cagcagcacg
      841 gggccgtcgc cgatgggggt gttctgctgg tagtggtcgg cgagctgcac gccgccgtcc
      901 tcgatgttgt ggcggatctt gaagttggcc ttgatgccgt tcttctgctt gtcggcggtg
      961 atatagacgt tgtggctgtt gtagttgtac tccagcttgt gccccaggat gttgccgtcc
     1021 tccttgaagt cgatgccctt cagctcgatg cggttcacca gggtgtcgcc ctcgaacttc
     1081 acctcggcgc gggtcttgta gttgccgtcg tccttgaaga agatggtgcg ctcctggacg
     1141 tagccttcgg gcatggcgga cttgaagaag tcgtgctgct tcatgtggtc ggggtagcgg
     1201 gcgaagcact gcacgccgta gcccagggtg gtcacgaggg tgggccaggg cacgggcagc
     1261 ttgccggtgg tgcagatcag cttcagggtc agcttgccgt aggtggcatc gccctcgccc
     1321 tcgccggaca cgctgaactt gtggccgttt acgtcgccgt ccagctcgac caggatgggc
     1381 accaccccgg tgaacagctc ctcgcccttg ctcaccatgg tggcgaccgg tagcgctagc
     1441 ttcgaaagat ctggatccca gaagcaaatg taagcaatag atggctctgc cctgactttt
     1501 atgcccagcc cgagctccga gttttgagac cagcctgggc aacataccga gatcctgtct
     1561 taaacgaatg aggagttagg agttagtaga gggcttgcct agcatacaca gagacctgag
     1621 attaatccca gcaccaagca aactgagggt gctggctccc acctgtaatc ccatactcag
     1681 gaggtgaggt cagcctgggc tacatgaaac cctgtagccc ctcagggctc tgagatggtc
     1741 ctgtagatat ggagcgtcct cagagagcga gtggacagct tggtccttgg tgctttattt
     1801 gggggtctgg gggaggctga cccagttacc cagccgcacc ctgtctgttc ctcacagctc
     1861 ggtctgagag atgagcatat ttcctttttc acacgttgcc tccacacttt gacaagtcta
     1921 ggcataatca gattgtctgt tttctaagta tggtgacaag aataggactt ttagttcctc
     1981 atcttcaaac caccaaggga aaaaagcctt cagagcattg cctagaaagt taacagttac
     2041 gaacacgatg ccaggacacc gcatttaaat ttttatgtgt acatttttgt caagcacctt
     2101 ggaagtttat agcatggaaa atgaatctgc cactaacttc agatctttaa cgattattac
     2161 tcttacaacg cgtgcggccg caggaacccc tagtgatgga gttggccact ccctctctgc
     2221 gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggcggcctca
     2281 gtgagcgagc gagcgcgcag ctgcctgcag gacatgtgag caaaaggcca gcaaaaggcc
     2341 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     2401 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     2461 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     2521 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     2581 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     2641 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     2701 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     2761 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     2821 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     2881 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     2941 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     3001 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     3061 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     3121 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     3181 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     3241 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     3301 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     3361 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     3421 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     3481 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     3541 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     3601 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     3661 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     3721 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     3781 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     3841 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     3901 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     3961 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     4021 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     4081 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     4141 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt
     4201 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt
     4261 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg
     4321 tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccataaa
     4381 attgtaaacg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     4441 tttaaccaat agaccgaaat cggcaaaatc ccttataaat caaaagaata gcccgagata
     4501 gagttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     4561 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcacccaaa
     4621 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     4681 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     4741 aaaggagcgg gcgctaaggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     4801 cccgccgcgc ttaatgcgcc gctacagggc gcgtactatg gttgctttga cgtatgcggt
     4861 gtgaaatacc gcacag
//
