LOCUS       sequence-449745-        5165 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0353h is an enhancer sequence, designed to drive AAV-mediated 
            transgene expression in cortical Pvalb+ neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5165)
  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 5165)
  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: AiP14521 -
            pAAV-AiE0353h-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5165
                     /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      518..527
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(524..526,527..529,530..1240)
                     /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(830..849)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1177..1198
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1250..1306
                     /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             1319..1384
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1415..1792
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(1879..1899)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2086..2197
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2315..2455
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2315..2444
                     /label=AAV2 ITR
     misc_feature    2462..2489
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     misc_feature    2490..2517
                     /gene="telL"
                     /label=telL
                     /note="telomeric sequence from coliphage N15"
     rep_origin      2586..3041
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2673..2692)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2883..2904
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3058..3077)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3177..3199
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3237..3255)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3323..3427
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3428..3496,3497..4288)
                     /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(3646..3665)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4459..5047
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4948..4967
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5104..5131
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     misc_feature    5132..5159
                     /gene="telL"
                     /label=telL
                     /note="telomeric sequence from coliphage N15"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtcctttc ttgctctatg tatttgagcc tagaaaaaga
      181 cacactccca agaaattaga ataaagggaa aaatgacttt gagatgtact ttcttaaaat
      241 aactgaagta atgaccgcat gagccagctg atgggttttt aaatggatac gtttctatca
      301 gcctgctgct tgatgccaga gccaaatata tatggagtgt attatatcat atcctgtagg
      361 caggagactg ttggggcatt gggggactta gagaggtgaa gtggcaggct tggcacagga
      421 attagagctc gggctgggca taaaagtcag ggcagagcca tctattgctt acatttgctt
      481 ctgggatcca gatctttcga agctagcgct accggtcgcc accatggtct ccaagggcga
      541 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgatg tcaacggcca
      601 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
      661 gctgatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctggg
      721 ctacggcgtg cagtgcttcg cccgctaccc cgaccacatg aagcagcacg acttcttcaa
      781 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaaag acgacggcaa
      841 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
      901 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
      961 caacagccac aacgtctata tcaccgccga caagcagaag aacggcatca aggccaactt
     1021 caagatccgc cacaacatcg aggacggcgg cgtgcagctc gccgaccact accagcagaa
     1081 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gctaccagtc
     1141 caagctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
     1201 cgccgccggg atcactctcg gcatggacga gctgtacaag ggctctggtg ctaccaactt
     1261 ctcactgttg aaacaggcag gggatgtaga ggagaatcca gggcctggtg ctagtggaga
     1321 ctacaaagac catgacggag attataaaga tcatgacatc gattacaagg atgacgatga
     1381 caagtccgga ctcagatctg gaggctccgg aggcccagag ccagcgaagt ctgctcccgc
     1441 cccgaaaaag ggctccaaga aggcggtgac taaggcgcag aagaaaggcg gcaagaagcg
     1501 caagcgcagc cgcaaggaga gctattccat ctatgtgtat aaggttctga agcaggtcca
     1561 ccctgacacc ggcatttcgt ccaaggccat gggcatcatg aactcgtttg tgaacgacat
     1621 tttcgagcgc atcgcaggtg aggcttcccg cctggcgcat tacaacaagc gctcgaccat
     1681 cacctccagg gagatccaga cggccgtgcg cctgctgctg cctggggagt tggccaagca
     1741 cgccgtgtcc gagggtacta aggccatcac caagtacacc agcgctaagt aatgaggcgc
     1801 gccgcggccg cgaattcgat atcataatca acctctggat tacaaaattt gtgaaagatt
     1861 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     1921 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     1981 gttagttctt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     2041 tcggctgttg ggcactgaca attccgtggc tcgagagatc ttcgactgtg ccttctagtt
     2101 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     2161 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     2221 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     2281 ggcatgagat ctcacgtgcg gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc
     2341 actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc
     2401 ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc
     2461 ctatcagcac acaattgccc attatacgcg cgtataatgg actattgtgt gctgatatga
     2521 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc
     2581 atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     2641 gaccgctaca cttgccagcg ccttagcgcc cgctcctttc gctttcttcc cttcctttct
     2701 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     2761 atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag
     2821 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     2881 tagtggactc ttgttccaaa ctggaacaac actcaactct atctcgggct attcttttga
     2941 tttataaggg attttgccga tttcggtcta ttggttaaaa aatgagctga tttaacaaaa
     3001 atttaacgcg aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac
     3061 aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc
     3121 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
     3181 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct
     3241 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     3301 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     3361 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     3421 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     3481 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     3541 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     3601 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     3661 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     3721 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     3781 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     3841 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     3901 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     3961 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     4021 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     4081 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     4141 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     4201 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     4261 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     4321 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     4381 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     4441 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     4501 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     4561 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     4621 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     4681 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     4741 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     4801 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     4861 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     4921 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     4981 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     5041 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     5101 tcatatcagc acacaattgc ccattatacg cgcgtataat ggactattgt gtgctgataa
     5161 catgt
//
