LOCUS       sequence-433114-        4578 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2572m 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 4578)
  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 4578)
  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: AiP1885 - pAAV-AiE2572m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4578
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     156..175
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   337..477
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   348..477
                     /label=AAV2 ITR
     polyA_signal    595..706
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     893..913
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(1006..1716,1717..1719,1720..1722))
                     /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(1048..1069)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1397..1416
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1656..1677
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1719..1728
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   2188..2317
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(2379..2967)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2459..2478)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3138..3929,3930..3998))
                     /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     3761..3780
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3999..4103)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4171..4189
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4227..4249)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4349..4368
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(join(4385..4578,1..262))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4522..4543)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 aaaaccgtct atcagggcga tggcccacta cgtgaaccat cacccaaatc aagttttttg
       61 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
      121 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
      181 gctaaggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
      241 aatgcgccgc tacagggcgc gtactatggt tgctttgacg tatgcggtgt gaaataccgc
      301 acagatgcgt aaggagaaaa taccgcatca ggcgcccctg caggcagctg cgcgctcgct
      361 cgctcactga ggccgcccgg gcaaagcccg ggcgtcgggc gacctttggt cgcccggcct
      421 cagtgagcga gcgagcgcgc agagagggag tggccaactc catcactagg ggttcctgcg
      481 gccgctcggt ccgcacgtga gatctcatgc ctgctattgt cttcccaatc ctcccccttg
      541 ctgtcctgcc ccaccccacc ccccagaata gaatgacacc tactcagaca atgcgatgca
      601 atttcctcat tttattagga aaggacagtg ggagtggcac cttccagggt caaggaaggc
      661 acgggggagg ggcaaacaac agatggctgg caactagaag gcacagtcga agatctctcg
      721 agccacggaa ttgtcagtgc ccaacagccg agcccctgtc cagcagcggg caaggcaggc
      781 ggcgatgagt tccgccgtgg caagaactaa ccaggattta tacaaggagg agaaaatgaa
      841 agccatacgg gaagcaatag catgatacaa aggcattaaa gcagcgtatc cacatagcgt
      901 aaaaggagca acatagttaa gaataccagt caatctttca caaattttgt aatccagagg
      961 ttgattatga tatcgaattc gcggccgcgg cgcgccgtcg acttacttgt acagctcgtc
     1021 catgccgaga gtgatcccgg cggcggtcac gaactccagc aggaccatgt gatcgcgctt
     1081 ctcgttgggg tctttgctca gcttggactg gtagctcagg tagtggttgt cgggcagcag
     1141 cacggggccg tcgccgatgg gggtgttctg ctggtagtgg tcggcgagct gcacgccgcc
     1201 gtcctcgatg ttgtggcgga tcttgaagtt ggccttgatg ccgttcttct gcttgtcggc
     1261 ggtgatatag acgttgtggc tgttgtagtt gtactccagc ttgtgcccca ggatgttgcc
     1321 gtcctccttg aagtcgatgc ccttcagctc gatgcggttc accagggtgt cgccctcgaa
     1381 cttcacctcg gcgcgggtct tgtagttgcc gtcgtccttg aagaagatgg tgcgctcctg
     1441 gacgtagcct tcgggcatgg cggacttgaa gaagtcgtgc tgcttcatgt ggtcggggta
     1501 gcgggcgaag cactgcacgc cgtagcccag ggtggtcacg agggtgggcc agggcacggg
     1561 cagcttgccg gtggtgcaga tcagcttcag ggtcagcttg ccgtaggtgg catcgccctc
     1621 gccctcgccg gacacgctga acttgtggcc gtttacgtcg ccgtccagct cgaccaggat
     1681 gggcaccacc ccggtgaaca gctcctcgcc cttgctcacc atggtggcga ccggtagcgc
     1741 tagcttcgaa agatctggat cccagaagca aatgtaagca atagatggct ctgccctgac
     1801 ttttatgccc agcccgagct ccgcccttat attcagcagg ccttggagct ggtacaagat
     1861 ggatattctg cccacactgt gctcgaaggt gtttcttgct tagcagtgtc ctgggtggga
     1921 ggttaagtga gtggtgattg gctcagaatt tgggattctt ctgtgttctc atccctctcg
     1981 gcagctaagg tgtttattaa aaatttggct gatttcttgg tgattacata tgtcccactg
     2041 ctctgtctgt ctgtgcctag aatacaacat cccatctcct ttcacctggg aatttagtct
     2101 ctttgagcat tgctgagtcc tctctagtag ttttaaataa aattgtgatg ttaaaagttt
     2161 acttggtgtt tttctttcat cggccgcagg aacccctagt gatggagttg gccactccct
     2221 ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggcg
     2281 gcctcagtga gcgagcgagc gcgcagctgc ctgcaggaca tgtgagcaaa aggccagcaa
     2341 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     2401 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     2461 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     2521 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     2581 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     2641 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     2701 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     2761 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     2821 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     2881 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     2941 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     3001 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     3061 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     3121 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     3181 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     3241 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     3301 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     3361 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     3421 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     3481 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     3541 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     3601 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     3661 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     3721 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     3781 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     3841 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     3901 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     3961 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     4021 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     4081 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     4141 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc
     4201 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
     4261 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
     4321 ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac
     4381 cataaaattg taaacgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     4441 tcatttttta accaatagac cgaaatcggc aaaatccctt ataaatcaaa agaatagccc
     4501 gagatagagt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     4561 tccaacgtca aagggcga
//
