LOCUS       sequence-449734-        5103 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0552h is an enhancer sequence, designed to drive AAV-mediated 
            transgene expression in cortical L5_IT and L6_IT neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5103)
  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 5103)
  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: AiP13655 -
            pAAV-AiE0552h-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5103
                     /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      569..578
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(575..577,578..580,581..1291)
                     /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(881..900)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1228..1249
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1301..1357
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             1370..1435
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1466..1843
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(1930..1950)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2137..2248
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2366..2506
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2366..2495
                     /label=AAV2 ITR
     rep_origin      2581..3036
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2668..2687)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2878..2899
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3053..3072)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3172..3194
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3232..3250)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3318..3422
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3423..3491,3492..4283)
                     /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(3641..3660)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4454..5042
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4943..4962
                     /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 gcggccgcac gcgtaagcca tgttattgca agagcactga gagacaaact
      181 tttaaaaaag cagatcgcat ggcaagtcca taaaatcagg attacaaagg catctggaca
      241 cagacactat gaattctaga tctgaacggg aatagttaag ttcgggaaga gacaattgtc
      301 agaattcatg ggcttagaac agtggcctgc acagctccct ggcacgtggc tagatgaagc
      361 tggattaaaa ataaagatca acccaaacct atagcaaacc agactaaaca tatggacaga
      421 aattttatgg aaaaagaagt aaaaatatct cttagtcata tgaaaatata tttcagagct
      481 cgggctgggc ataaaagtca gggcagagcc atctattgct tacatttgct tctgggatcc
      541 agatctttcg aagctagcgc taccggtcgc caccatggtc tccaagggcg aggagctgtt
      601 caccggggtg gtgcccatcc tggtcgagct ggacggcgat gtcaacggcc acaagttcag
      661 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agctgatctg
      721 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctgg gctacggcgt
      781 gcagtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
      841 gcccgaaggc tacgtccagg agcgcaccat cttcttcaaa gacgacggca actacaagac
      901 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
      961 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca
     1021 caacgtctat atcaccgccg acaagcagaa gaacggcatc aaggccaact tcaagatccg
     1081 ccacaacatc gaggacggcg gcgtgcagct cgccgaccac taccagcaga acacccccat
     1141 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agctaccagt ccaagctgag
     1201 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
     1261 gatcactctc ggcatggacg agctgtacaa gggctctggt gctaccaact tctcactgtt
     1321 gaaacaggca ggggatgtag aggagaatcc agggcctggt gctagtggag actacaaaga
     1381 ccatgacgga gattataaag atcatgacat cgattacaag gatgacgatg acaagtccgg
     1441 actcagatct ggaggctccg gaggcccaga gccagcgaag tctgctcccg ccccgaaaaa
     1501 gggctccaag aaggcggtga ctaaggcgca gaagaaaggc ggcaagaagc gcaagcgcag
     1561 ccgcaaggag agctattcca tctatgtgta caaggttctg aagcaggtcc accctgacac
     1621 cggcatttcg tccaaggcca tgggcatcat gaattcgttt gtgaacgaca ttttcgagcg
     1681 catcgcagga gaggcttccc gcctggcgca ttacaacaag cgctcgacca tcacctcccg
     1741 ggagatccag acggccgtgc gcctgctgct gcctggggag ttggccaagc acgccgtgtc
     1801 cgagggtact aaggccatca ccaagtacac cagcgctaag taatgaggcg cgccgcggcc
     1861 gcgaattcga tatcataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     1921 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     1981 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttagttct
     2041 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     2101 gggcactgac aattccgtgg ctcgagagat cttcgactgt gccttctagt tgccagccat
     2161 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     2221 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     2281 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgaga
     2341 tctcacgtgc ggaccgagcg gccgcaggaa cccctagtga tggagttggc cactccctct
     2401 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt
     2461 gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg
     2521 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt
     2581 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     2641 ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     2701 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     2761 gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc
     2821 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     2881 gactcttgtt ccaaactgga acaacactca actctatctc gggctattct tttgatttat
     2941 aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta
     3001 acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct
     3061 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct
     3121 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     3181 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga
     3241 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     3301 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     3361 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     3421 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     3481 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     3541 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     3601 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     3661 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     3721 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     3781 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     3841 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     3901 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     3961 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     4021 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     4081 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     4141 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     4201 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     4261 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     4321 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     4381 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     4441 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     4501 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     4561 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     4621 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     4681 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     4741 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     4801 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     4861 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     4921 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     4981 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     5041 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     5101 tgt
//
