LOCUS       sequence-470533-        4538 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0660m 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 4538)
  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 4538)
  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: AiP12851 - pAAV-AiE0660m-minBG-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..4538
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      163..172
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(169..171,172..174,175..885)
                     /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(214..235)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(475..494)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     822..843
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(978..998)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1185..1296
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1414..1554
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1414..1543
                     /label=AAV2 ITR
     rep_origin      1629..2084
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1716..1735)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1926..1947
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2101..2120)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2220..2242
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2280..2298)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2366..2470
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2471..2539,2540..3331)
                     /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(2689..2708)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3502..4090
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3991..4010
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   4152..4281
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 aatattattt gtggcctact agagactgag cctaaggcta taaacatgtt agtcgtgctc
       61 taccagtgag agctcgggct gggcataaaa gtcagggcag agccatctat tgcttacatt
      121 tgcttctggg atccagatct ttcgaagcta gcgctaccgg tcgccaccat ggtgagcaag
      181 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
      241 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
      301 ctgaagctga tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
      361 ctgggctacg gcgtgcagtg cttcgcccgc taccccgacc acatgaagca gcacgacttc
      421 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
      481 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
      541 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
      601 aactacaaca gccacaacgt ctatatcacc gccgacaagc agaagaacgg catcaaggcc
      661 aacttcaaga tccgccacaa catcgaggac ggcggcgtgc agctcgccga ccactaccag
      721 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagctac
      781 cagtccaagc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
      841 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaagt cgacggcgcg
      901 ccgcggccgc gaattcgata tcataatcaa cctctggatt acaaaatttg tgaaagattg
      961 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     1021 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     1081 ttagttcttg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
     1141 cggctgttgg gcactgacaa ttccgtggct cgagagatct tcgactgtgc cttctagttg
     1201 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     1261 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     1321 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
     1381 gcatgagatc tcacgtgcgg accgagcggc cgcaggaacc cctagtgatg gagttggcca
     1441 ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc
     1501 cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc
     1561 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata cgtcaaagca
     1621 accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     1681 cgtgaccgct acacttgcca gcgccttagc gcccgctcct ttcgctttct tcccttcctt
     1741 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     1801 ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg
     1861 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     1921 taatagtgga ctcttgttcc aaactggaac aacactcaac tctatctcgg gctattcttt
     1981 tgatttataa gggattttgc cgatttcggt ctattggtta aaaaatgagc tgatttaaca
     2041 aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt gcactctcag
     2101 tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga
     2161 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     2221 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg
     2281 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     2341 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     2401 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     2461 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     2521 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     2581 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     2641 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     2701 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     2761 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     2821 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     2881 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     2941 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     3001 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     3061 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     3121 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     3181 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     3241 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     3301 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     3361 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     3421 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     3481 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     3541 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     3601 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     3661 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     3721 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     3781 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     3841 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     3901 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     3961 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     4021 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     4081 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     4141 tgctcacatg tcctgcaggc agctgcgcgc tcgctcgctc actgaggccg cccgggcgtc
     4201 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
     4261 actccatcac taggggttcc tgcggccgca cgcgttactg tccaggccag ctttgaacag
     4321 actctgtatt ccagaaagac tttcagcttg tgcttctttt ccccaaacct gccttgagta
     4381 gctgacatca taggcttgca ctaccaggcc tggcttaaaa tacatactcc caatttacat
     4441 atctgatgag taacattctc atagaagcac aaaagtacat acatggattc taatataggt
     4501 tattttcttg ggcattacac actacagata tactattc
//
