LOCUS       sequence-432579-        4706 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2570m 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 4706)
  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 4706)
  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: AiP1879 - pAAV-AiE2570m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4706
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   137..277
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   148..277
                     /label=AAV2 ITR
     polyA_signal    395..506
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     693..713
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(806..1516,1517..1519,1520..1522))
                     /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(848..869)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1197..1216
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1456..1477
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1519..1528
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   2116..2245
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(2307..2895)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2387..2406)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3066..3857,3858..3926))
                     /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     3689..3708
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3927..4031)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4099..4117
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4155..4177)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4277..4296
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(join(4313..4706,1..62))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4450..4471)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4662..4681
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
       61 gtactatggt tgctttgacg tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa
      121 taccgcatca ggcgcccctg caggcagctg cgcgctcgct cgctcactga ggccgcccgg
      181 gcaaagcccg ggcgtcgggc gacctttggt cgcccggcct cagtgagcga gcgagcgcgc
      241 agagagggag tggccaactc catcactagg ggttcctgcg gccgctcggt ccgcacgtga
      301 gatctcatgc ctgctattgt cttcccaatc ctcccccttg ctgtcctgcc ccaccccacc
      361 ccccagaata gaatgacacc tactcagaca atgcgatgca atttcctcat tttattagga
      421 aaggacagtg ggagtggcac cttccagggt caaggaaggc acgggggagg ggcaaacaac
      481 agatggctgg caactagaag gcacagtcga agatctctcg agccacggaa ttgtcagtgc
      541 ccaacagccg agcccctgtc cagcagcggg caaggcaggc ggcgatgagt tccgccgtgg
      601 caagaactaa ccaggattta tacaaggagg agaaaatgaa agccatacgg gaagcaatag
      661 catgatacaa aggcattaaa gcagcgtatc cacatagcgt aaaaggagca acatagttaa
      721 gaataccagt caatctttca caaattttgt aatccagagg ttgattatga tatcgaattc
      781 gcggccgcgg cgcgccgtcg acttacttgt acagctcgtc catgccgaga gtgatcccgg
      841 cggcggtcac gaactccagc aggaccatgt gatcgcgctt ctcgttgggg tctttgctca
      901 gcttggactg gtagctcagg tagtggttgt cgggcagcag cacggggccg tcgccgatgg
      961 gggtgttctg ctggtagtgg tcggcgagct gcacgccgcc gtcctcgatg ttgtggcgga
     1021 tcttgaagtt ggccttgatg ccgttcttct gcttgtcggc ggtgatatag acgttgtggc
     1081 tgttgtagtt gtactccagc ttgtgcccca ggatgttgcc gtcctccttg aagtcgatgc
     1141 ccttcagctc gatgcggttc accagggtgt cgccctcgaa cttcacctcg gcgcgggtct
     1201 tgtagttgcc gtcgtccttg aagaagatgg tgcgctcctg gacgtagcct tcgggcatgg
     1261 cggacttgaa gaagtcgtgc tgcttcatgt ggtcggggta gcgggcgaag cactgcacgc
     1321 cgtagcccag ggtggtcacg agggtgggcc agggcacggg cagcttgccg gtggtgcaga
     1381 tcagcttcag ggtcagcttg ccgtaggtgg catcgccctc gccctcgccg gacacgctga
     1441 acttgtggcc gtttacgtcg ccgtccagct cgaccaggat gggcaccacc ccggtgaaca
     1501 gctcctcgcc cttgctcacc atggtggcga ccggtagcgc tagcttcgaa agatctggat
     1561 cccagaagca aatgtaagca atagatggct ctgccctgac ttttatgccc agcccgagct
     1621 ccgttccggt ttgctgctga aactgtgtaa cgggtaacac ctcacctcct tctcaatctt
     1681 ccattggttc ctttgtggct accaaaaagt tctaaagttt ccaaaaccca ctcggtctgt
     1741 tcaacattgt gcccgtgacc atcatttgac aggcaggagg accacagcac ttctgagttc
     1801 tgcgcatgcc cagtgcatcc ttcctgttct ggggttccct ctctgctccg tcctgtgagc
     1861 ttatcaacag aaaacatcga gcactgcttg gtagcacttc agttccacct tggcttattg
     1921 gttacttttt tctcactttg tgcagaaaag caacttaagg aaggaaggac ttcttctggc
     1981 ttgcagttgg agagttagtc cgtcgtgaca agttagacta cagcagctca attagagcgt
     2041 tggcgacaga aagaagtgtg tgttgtctgg tcacagagac atatctccta ggtgattcta
     2101 aatccatgcg gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc
     2161 gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcggc ctcagtgagc
     2221 gagcgagcgc gcagctgcct gcaggacatg tgagcaaaag gccagcaaaa ggccaggaac
     2281 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     2341 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     2401 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     2461 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     2521 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     2581 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     2641 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     2701 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     2761 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     2821 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     2881 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     2941 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     3001 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     3061 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     3121 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     3181 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     3241 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     3301 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     3361 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     3421 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     3481 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     3541 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     3601 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     3661 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     3721 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     3781 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     3841 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     3901 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     3961 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     4021 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
     4081 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt
     4141 gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa
     4201 gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg
     4261 ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca taaaattgta
     4321 aacgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     4381 caatagaccg aaatcggcaa aatcccttat aaatcaaaag aatagcccga gatagagttg
     4441 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     4501 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc caaatcaagt
     4561 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     4621 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     4681 gcgggcgcta aggcgctggc aagtgt
//
