LOCUS       sequence-474062-        5548 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0174h 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 5548)
  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 5548)
  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: AiP11667 -
            pAAV-hsA2-AiE0174h-minRho-SYFP2-WPR...
FEATURES             Location/Qualifiers
     source          1..5548
                     /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      1585..1594
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1591..1593,1594..1596,1597..2307)
                     /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(1636..1657)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1897..1916)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2244..2265
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2375..2395)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2582..2693
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2811..2951
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2811..2940
                     /label=AAV2 ITR
     rep_origin      3026..3481
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3113..3132)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3323..3344
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3498..3517)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3617..3639
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3677..3695)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3763..3867
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3868..3936,3937..4728)
                     /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(4086..4105)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4899..5487
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5388..5407
                     /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 tttgttttgc
      361 taccctaaat attccctgtg ctccatctat tcatttctcc tcccttccat cttgaaacca
      421 ctgatctttc tgcttcctct ataattttgc cttttccttt tccagaatgt tataagggtg
      481 gaatcatata gtgtgcagcc ttgaaagagt ggcttcttcc accaaataat gtgcatttaa
      541 ggttccttca tgtctttaca tggcttgata gctcactttt tatccctgaa taataactca
      601 ttatacagat gtatcaccat ttgtttaccc attcatctgt tgaagggcat gttgattgct
      661 tccagctttt gtcaattatg gataggctgc tataaacatt catgtgtagg cttttaccaa
      721 cacatttgga caaactccaa ggagcacaat tgctggatta tgtaagtaga ctatgtttca
      781 ttacgtaaga agctttcaac ctgtcttgta aagtggttct atcattttgc attcccacta
      841 gcaataaagg agagttgttc tacatttttg gaacatttgg tgttgtcata ttgggcaagg
      901 ggtattttag ctattctaaa agggatctca taagtttaat ttgcaatatc ctaataaaat
      961 atgatgttga acatcttttc atacgattat ttgctatttg tatattgtct ttgatgaggt
     1021 gcctactcag acattttccc cactgttgaa atggattttt tgttttccaa tttgtttcat
     1081 gtggacaaaa ttataattta tgttatattg ccttgtagaa tgctgtgtag ataatgtaac
     1141 aggctaagga atagaaataa tttgatggaa aaacaaatct actttttgtt ttacaaagaa
     1201 atgttatgaa agccctttct gaggagaaag cattttactt gaaaattttt taatagaggc
     1261 agggtcttgt tctgttcccc aggttggagt gcagtgacac aaccatagct cattgcagcc
     1321 ttgaactcct gagctcaagc gatcctccag agtagctagc actttaggct agagccacta
     1381 taacccagcc aggaattcga tatcataatc aaccataggt accgagctcg ggattcagcc
     1441 gggagcttag ggaggggagg tcacttcata agggcctggg gggggagttg gagccacgag
     1501 tcgtccagcc ggagccccgt gtggctgagc tccggcctca gaagcatccc cgggttggat
     1561 ccttcgaagc tagcgctacc ggtcgccacc atggtgagca agggcgagga gctgttcacc
     1621 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
     1681 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagct gatctgcacc
     1741 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgggcta cggcgtgcag
     1801 tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     1861 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
     1921 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     1981 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac
     2041 gtctatatca ccgccgacaa gcagaagaac ggcatcaagg ccaacttcaa gatccgccac
     2101 aacatcgagg acggcggcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     2161 gacggccccg tgctgctgcc cgacaaccac tacctgagct accagtccaa gctgagcaaa
     2221 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     2281 actctcggca tggacgagct gtacaagtaa gtcgacatca taatcaacct ctggattaca
     2341 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat
     2401 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct
     2461 ccttgtataa atcctggtta gttcttgcca cggcggaact catcgccgcc tgccttgccc
     2521 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggctcga gagatcttcg
     2581 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     2641 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     2701 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     2761 tgggaagaca atagcaggca tgagatctca cgtgcggacc gagcggccgc aggaacccct
     2821 agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc
     2881 aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag
     2941 ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     3001 ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt
     3061 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc cgctcctttc
     3121 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     3181 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     3241 ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     3301 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaactct
     3361 atctcgggct attcttttga tttataaggg attttgccga tttcggtcta ttggttaaaa
     3421 aatgagctga tttaacaaaa atttaacggg aattttaaca aaatattaac gtttacaatt
     3481 ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac
     3541 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     3601 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     3661 cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     3721 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     3781 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     3841 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     3901 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     3961 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     4021 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     4081 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     4141 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     4201 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     4261 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     4321 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     4381 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     4441 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     4501 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     4561 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     4621 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     4681 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     4741 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     4801 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     4861 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     4921 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     4981 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     5041 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     5101 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     5161 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     5221 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     5281 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     5341 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     5401 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     5461 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     5521 gccttttgct ggccttttgc tcacatgt
//
