LOCUS       sequence-470487-        4559 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0758m 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 4559)
  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 4559)
  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: AiP12640 - pAAV-AiE0758m-minBG-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..4559
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      603..1191
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1092..1111
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   1253..1382
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      1823..1832
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1829..1831,1832..1834,1835..2545)
                     /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(1874..1895)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2135..2154)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2482..2503
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2638..2658)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2845..2956
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3074..3214
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3074..3203
                     /label=AAV2 ITR
     rep_origin      3289..3744
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3376..3395)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3586..3607
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3761..3780)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3880..3902
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3940..3958)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4026..4130
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4131..4199,4200..4559,1..432)
                     /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(4349..4368)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
       61 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
      121 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
      181 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
      241 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
      301 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
      361 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
      421 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
      481 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
      541 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
      601 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
      661 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
      721 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
      781 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
      841 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
      901 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
      961 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     1021 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     1081 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     1141 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     1201 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gtcctgcagg
     1261 cagctgcgcg ctcgctcgct cactgaggcc gcccgggcgt cgggcgacct ttggtcgccc
     1321 ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc aactccatca ctaggggttc
     1381 ctgcggccgc acgcgtagga gactcaggga tgcacaaagg caccgttgaa tgtcttcgga
     1441 ggatgatttg gcgagaggac tgtaaagcct ggatttacaa atattacaca agtgaacact
     1501 gttactgcac tcaggctgca aaaccctgct caaatccaat aagtggttta agatgtgcaa
     1561 atagaggctt ctgcggggag tcaggcatta gaagccacct ggagcatgtg gtagagattg
     1621 tatccatcaa tacagcagct gggctgaaat tataaagaga gctgcttttc agcacgcttg
     1681 ttttaatgca tacatttctt ctgtcactgg caccccagga aaactactag agctcgggct
     1741 gggcataaaa gtcagggcag agccatctat tgcttacatt tgcttctggg atccagatct
     1801 ttcgaagcta gcgctaccgg tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg
     1861 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc
     1921 cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagctga tctgcaccac
     1981 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgggctacg gcgtgcagtg
     2041 cttcgcccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
     2101 aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc
     2161 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt
     2221 caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt
     2281 ctatatcacc gccgacaagc agaagaacgg catcaaggcc aacttcaaga tccgccacaa
     2341 catcgaggac ggcggcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga
     2401 cggccccgtg ctgctgcccg acaaccacta cctgagctac cagtccaagc tgagcaaaga
     2461 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
     2521 tctcggcatg gacgagctgt acaagtaagt cgacggcgcg ccgcggccgc gaattcgata
     2581 tcataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     2641 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     2701 ccgtatggct ttcattttct cctccttgta taaatcctgg ttagttcttg ccacggcgga
     2761 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     2821 ttccgtggct cgagagatct tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     2881 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     2941 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     3001 ggcaggacag caagggggag gattgggaag acaatagcag gcatgagatc tcacgtgcgg
     3061 accgagcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc
     3121 tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc
     3181 tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt
     3241 acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta
     3301 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     3361 gcgccttagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     3421 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     3481 acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat
     3541 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     3601 aaactggaac aacactcaac tctatctcgg gctattcttt tgatttataa gggattttgc
     3661 cgatttcggt ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     3721 acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg
     3781 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     3841 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     3901 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt
     3961 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     4021 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     4081 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     4141 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     4201 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     4261 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     4321 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     4381 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     4441 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     4501 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccg
//
