LOCUS       sequence-470501-        4573 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0648h 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 4573)
  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 4573)
  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: AiP12686 - pAAV-AiE0648h-minBG-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..4573
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(335..1126,1127..1195))
                     /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     958..977
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1196..1300)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1368..1386
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1424..1446)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1546..1565
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(1582..2037)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1719..1740)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1931..1950
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2112..2252
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2123..2252
                     /label=AAV2 ITR
     polyA_signal    2370..2481
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2668..2688
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(2781..3491,3492..3494,3495..3497))
                     /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(2823..2844)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3172..3191
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3431..3452
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3494..3503
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   3958..4087
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(join(4149..4573,1..164))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4229..4248)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
       61 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
      121 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
      181 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
      241 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
      301 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
      361 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
      421 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
      481 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
      541 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
      601 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
      661 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
      721 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
      781 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
      841 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
      901 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
      961 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     1021 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     1081 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     1141 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     1201 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     1261 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     1321 cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
     1381 cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
     1441 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
     1501 gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga
     1561 ttgtactgag agtgcaccat aaaattgtaa acgttaatat tttgttaaaa ttcgcgttaa
     1621 atttttgtta aatcagctca ttttttaacc aatagaccga aatcggcaaa atcccttata
     1681 aatcaaaaga atagcccgag atagagttga gtgttgttcc agtttggaac aagagtccac
     1741 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     1801 cactacgtga accatcaccc aaatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     1861 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     1921 cgagaaagga agggaagaaa gcgaaaggag cgggcgctaa ggcgctggca agtgtagcgg
     1981 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtact
     2041 atggttgctt tgacgtatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
     2101 catcaggcgc ccctgcaggc agctgcgcgc tcgctcgctc actgaggccg cccgggcaaa
     2161 gcccgggcgt cgggcgacct ttggtcgccc ggcctcagtg agcgagcgag cgcgcagaga
     2221 gggagtggcc aactccatca ctaggggttc ctgcggccgc tcggtccgca cgtgagatct
     2281 catgcctgct attgtcttcc caatcctccc ccttgctgtc ctgccccacc ccacccccca
     2341 gaatagaatg acacctactc agacaatgcg atgcaatttc ctcattttat taggaaagga
     2401 cagtgggagt ggcaccttcc agggtcaagg aaggcacggg ggaggggcaa acaacagatg
     2461 gctggcaact agaaggcaca gtcgaagatc tctcgagcca cggaattgtc agtgcccaac
     2521 agccgagccc ctgtccagca gcgggcaagg caggcggcga tgagttccgc cgtggcaaga
     2581 actaaccagg atttatacaa ggaggagaaa atgaaagcca tacgggaagc aatagcatga
     2641 tacaaaggca ttaaagcagc gtatccacat agcgtaaaag gagcaacata gttaagaata
     2701 ccagtcaatc tttcacaaat tttgtaatcc agaggttgat tatgatatcg aattcgcggc
     2761 cgcggcgcgc cgtcgactta cttgtacagc tcgtccatgc cgagagtgat cccggcggcg
     2821 gtcacgaact ccagcaggac catgtgatcg cgcttctcgt tggggtcttt gctcagcttg
     2881 gactggtagc tcaggtagtg gttgtcgggc agcagcacgg ggccgtcgcc gatgggggtg
     2941 ttctgctggt agtggtcggc gagctgcacg ccgccgtcct cgatgttgtg gcggatcttg
     3001 aagttggcct tgatgccgtt cttctgcttg tcggcggtga tatagacgtt gtggctgttg
     3061 tagttgtact ccagcttgtg ccccaggatg ttgccgtcct ccttgaagtc gatgcccttc
     3121 agctcgatgc ggttcaccag ggtgtcgccc tcgaacttca cctcggcgcg ggtcttgtag
     3181 ttgccgtcgt ccttgaagaa gatggtgcgc tcctggacgt agccttcggg catggcggac
     3241 ttgaagaagt cgtgctgctt catgtggtcg gggtagcggg cgaagcactg cacgccgtag
     3301 cccagggtgg tcacgagggt gggccagggc acgggcagct tgccggtggt gcagatcagc
     3361 ttcagggtca gcttgccgta ggtggcatcg ccctcgccct cgccggacac gctgaacttg
     3421 tggccgttta cgtcgccgtc cagctcgacc aggatgggca ccaccccggt gaacagctcc
     3481 tcgcccttgc tcaccatggt ggcgaccggt agcgctagct tcgaaagatc tggatcccag
     3541 aagcaaatgt aagcaataga tggctctgcc ctgactttta tgcccagccc gagctcatgg
     3601 caggaaaatg gaatgggctc ttccaagaag ttcccaaggc agctgaaaat attggagtta
     3661 agactgtggg gcctgcgttc aagtcctggc tctgcccata tgaactgact tggctttgaa
     3721 taaagccctt tttcctctct ctctctcttt tttttaatct cttttctgtc ctgtaaaatg
     3781 gacatctgac acctacctca tagtgttctt gtgagaattt catgaaacga tgcagaagaa
     3841 agacctacgt aagcctcaaa gcacaggagg aaacgcattc ctaatgctac tcatgctgct
     3901 gctgctgttg tggataatga taaaggccct taaagtggtg cctacgcgtg cggccgcagg
     3961 aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg
     4021 ggcgaccaaa ggtcgcccga cgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc
     4081 ctgcaggaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     4141 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     4201 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     4261 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     4321 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     4381 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     4441 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     4501 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     4561 tggtggccta act
//
