LOCUS       sequence-470444-        4726 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0398h 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 4726)
  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 4726)
  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: AiP12160 - pAAV-AiE0398h-minBG-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..4726
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     168..187
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   329..458
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      1066..1075
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1072..1074,1075..1077,1078..1788)
                     /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(1117..1138)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1378..1397)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1725..1746
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1881..1901)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2088..2199
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2317..2457
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2317..2446
                     /label=AAV2 ITR
     rep_origin      2532..2987
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2619..2638)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2829..2850
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3004..3023)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3123..3145
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3183..3201)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3269..3373
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3374..3442,3443..4234)
                     /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(3592..3611)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(4405..4726,1..267)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
       61 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
      121 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
      181 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
      241 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
      301 gccttttgct ggccttttgc tcacatgtcc tgcaggcagc tgcgcgctcg ctcgctcact
      361 gaggccgccc gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg
      421 cagagaggga gtggccaact ccatcactag gggttcctgc ggccgcacgc gtgctccggc
      481 ctctctattc ccactgctcc atccaggaaa accacgccca gcatggcaaa gcaaaccctt
      541 cctggggtcc ccctcactgg ggacacatct gggcaagcca aggacactca gaactcagcc
      601 tggccaggct cggtccccac ccccgctctg gcccttctcc ctacagtgaa agggacttta
      661 tggggacaaa aagccaccca ttgtatcaca cgagacaaag gggcagggga cagaggctcc
      721 gggcgcctaa gctggccacg acacagccac acaccacccc ctgactgtgt gagagggagg
      781 gcaccttgga attcccatgc catgcattcc agcattcgag gatcctggag tcatccccgc
      841 tctccaggaa ggcgacagca gactcggaac ccagccccag atccactaga cctgggcagt
      901 actgaatagc cctaagctct cttttctcgt cggaaaaatg ggcatgtaag ttgccaaacg
      961 gagtcattca gtgagctcgg gctgggcata aaagtcaggg cagagccatc tattgcttac
     1021 atttgcttct gggatccaga tctttcgaag ctagcgctac cggtcgccac catggtgagc
     1081 aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta
     1141 aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg
     1201 accctgaagc tgatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc
     1261 accctgggct acggcgtgca gtgcttcgcc cgctaccccg accacatgaa gcagcacgac
     1321 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac
     1381 gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
     1441 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag
     1501 tacaactaca acagccacaa cgtctatatc accgccgaca agcagaagaa cggcatcaag
     1561 gccaacttca agatccgcca caacatcgag gacggcggcg tgcagctcgc cgaccactac
     1621 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc
     1681 taccagtcca agctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag
     1741 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta agtcgacggc
     1801 gcgccgcggc cgcgaattcg atatcataat caacctctgg attacaaaat ttgtgaaaga
     1861 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
     1921 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
     1981 tggttagttc ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     2041 gctcggctgt tgggcactga caattccgtg gctcgagaga tcttcgactg tgccttctag
     2101 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     2161 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     2221 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     2281 caggcatgag atctcacgtg cggaccgagc ggccgcagga acccctagtg atggagttgg
     2341 ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac
     2401 gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc
     2461 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa
     2521 gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     2581 cagcgtgacc gctacacttg ccagcgcctt agcgcccgct cctttcgctt tcttcccttc
     2641 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     2701 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc
     2761 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     2821 ctttaatagt ggactcttgt tccaaactgg aacaacactc aactctatct cgggctattc
     2881 ttttgattta taagggattt tgccgatttc ggtctattgg ttaaaaaatg agctgattta
     2941 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct
     3001 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc
     3061 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     3121 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa
     3181 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     3241 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     3301 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     3361 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     3421 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     3481 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     3541 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     3601 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     3661 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     3721 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     3781 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     3841 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     3901 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     3961 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     4021 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     4081 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     4141 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     4201 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     4261 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     4321 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     4381 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     4441 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     4501 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     4561 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     4621 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     4681 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacg
//
