LOCUS       sequence-470896-        5261 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0089m 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 5261)
  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 5261)
  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: AiP11583 -
            pAAV-hsA2-AiE0089m-minRho-SYFP2-WPR...
FEATURES             Location/Qualifiers
     source          1..5261
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      1298..1307
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1304..1306,1307..1309,1310..2020)
                     /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(1349..1370)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1610..1629)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1957..1978
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2088..2108)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2295..2406
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2524..2664
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2524..2653
                     /label=AAV2 ITR
     rep_origin      2739..3194
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2826..2845)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3036..3057
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3211..3230)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3330..3352
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3390..3408)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3476..3580
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3581..3649,3650..4441)
                     /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     complement(3799..3818)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4612..5200
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5101..5120
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcaa cgcgtttaga acaatggctg gcccatagta aatgccgtgt
      181 tagtgtgtta gttgctgttc ttccacgtca gaagaggcac agacaaatta ccaccaggtg
      241 gcgctcagag tctgcggagg catcacaaca gccctgaatt tgaatcctgc tctgccactg
      301 cctagttgag accttttact acctgactag ctgtttgtgt attttaggtg tttgtttatg
      361 ggtgggcaag agacattctg aagtgagctg gtttccagga atatgtacaa acagctcaaa
      421 tggggataac ttagaagtgg cagtctgagg aaacagctgg atttgacagt taggaggtaa
      481 gagatgcaaa tcatgcattc acagtgcatg ctcctactct gagcatccaa cccgaggacc
      541 acgtgtggct gtggcctgga tgccagcgtg ctatgaaact gatgcttgta tgtgtgcgtg
      601 cacgagctct tcaagtgctt tatgcttctg gcttattcat gtggaaattt tcttaaagta
      661 atggattttg caagctttgc agaaaactga gtgcgtacac gagagccaca ttttctactg
      721 atcgtttgag ctatgatttg aaaagagtaa atatggcctt cttgccgcag ggtctgcttc
      781 ccctctttcc cttagacatt aaagtgactt gtaattacct ctcttaagct ggattctgga
      841 agaataacag tgattcgcaa gcacggtgaa gctctgctga aacccaggct gtatttaccg
      901 ctattgtctc cgagatcaga ccctcgctgg gcgagatagg agagaagaaa cggatagcac
      961 atttctaacc gacagtagca tttcttagga attgttatgg aaggagctgg aagcaaaatg
     1021 gttttcaaaa atataagatt gcggcataaa ttcaattttt aaaaagccgt ttagttccaa
     1081 taaaatcaga gtcttggtgc aaaaggaatt cgatatcata atcaaccata ggtaccgagc
     1141 tcgggattca gccgggagct tagggagggg aggtcacttc ataagggcct ggggggggag
     1201 ttggagccac gagtcgtcca gccggagccc cgtgtggctg agctccggcc tcagaagcat
     1261 ccccgggttg gatccttcga agctagcgct accggtcgcc accatggtga gcaagggcga
     1321 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
     1381 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
     1441 gctgatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctggg
     1501 ctacggcgtg cagtgcttcg cccgctaccc cgaccacatg aagcagcacg acttcttcaa
     1561 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa
     1621 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     1681 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
     1741 caacagccac aacgtctata tcaccgccga caagcagaag aacggcatca aggccaactt
     1801 caagatccgc cacaacatcg aggacggcgg cgtgcagctc gccgaccact accagcagaa
     1861 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gctaccagtc
     1921 caagctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
     1981 cgccgccggg atcactctcg gcatggacga gctgtacaag taagtcgaca tcataatcaa
     2041 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     2101 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     2161 ttcattttct cctccttgta taaatcctgg ttagttcttg ccacggcgga actcatcgcc
     2221 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggct
     2281 cgagagatct tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     2341 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     2401 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     2461 caagggggag gattgggaag acaatagcag gcatgagatc tcacgtgcgg accgagcggc
     2521 cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg
     2581 aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg
     2641 agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt acgcatctgt
     2701 gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt
     2761 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccttagc
     2821 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     2881 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     2941 caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     3001 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     3061 aacactcaac tctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggt
     3121 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gggaatttta acaaaatatt
     3181 aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     3241 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     3301 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     3361 gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     3421 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     3481 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     3541 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     3601 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     3661 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     3721 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     3781 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     3841 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     3901 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     3961 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     4021 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     4081 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     4141 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     4201 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     4261 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     4321 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     4381 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     4441 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     4501 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     4561 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     4621 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     4681 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     4741 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     4801 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     4861 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     4921 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     4981 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     5041 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     5101 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     5161 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     5221 tttacggttc ctggcctttt gctggccttt tgctcacatg t
//
