LOCUS       sequence-470709-        5558 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE1018h 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 5558)
  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 5558)
  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: AiP13610 -
            pAAV-AiE1018h-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5558
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      938..947
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(944..946,947..949,950..1660)
                     /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(1250..1269)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1597..1618
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1670..1726
                     /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             1739..1804
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1835..2212
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(2299..2319)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2506..2617
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2735..2875
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2735..2864
                     /label=AAV2 ITR
     rep_origin      2950..3405
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3037..3056)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3247..3268
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3422..3441)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3541..3563
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3601..3619)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3687..3791
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3792..3860,3861..4652)
                     /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(4010..4029)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4823..5411
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5312..5331
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   join(5473..5558,1..44)
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 agcgcgcaga gagggagtgg ccaactccat cactaggggt tcctgcggcc gcacgcgttc
       61 gccagtgcct tagtaggaat tataattaca tcaggagacc ccaggcaacc aagtaaactg
      121 attagctact gctccgagtt gatagatttt tttctctcta ggtcataaat agggctgtgg
      181 gagatggtgg aagcatgctg gtgggaacag gtcgagatgg gctgtacatt caggaagccc
      241 aaccaagtgg tgtgggcagc tccccctgcc ccaccctgcc accggaagag acagtctgac
      301 aggtgcccaa ggtagagttc agttccctcg gccagggact gcactcctga tcttgcgtgc
      361 tgcagagagc tgtcctcaga gcaggctgag ggtggctgaa ctgctaggcg atgctctggg
      421 aagccctccg ccaggaaagc cgatgctttc tgccaggcat aattattgca tggataatta
      481 cacgattcag agcctgctgg agaatcctgg gctggtgcca gagtagaaag gcagggttcc
      541 ggaactgcag ccagtggctg attctcagag tcgactttga aagaagaggg taatcattcc
      601 tcctcctaag actcctacct ggctgggcca ggacatggca aacgtaaagc taagggaggc
      661 ggggagttga tgagttctct cctccaccca ctcccttaca ggagggggtc cccagagttc
      721 tgacctaatc cctcttcttt actccctgca cgtgtcacct ccctgggaca atccacccct
      781 gcccattagc acatatatgc ggatgacacc aagtgtcttt ccagccaggc atctgcaaat
      841 gcatgagctc gggctgggca taaaagtcag ggcagagcca tctattgctt acatttgctt
      901 ctgggatcca gatctttcga agctagcgct accggtcgcc accatggtct ccaagggcga
      961 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgatg tcaacggcca
     1021 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
     1081 gctgatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctggg
     1141 ctacggcgtg cagtgcttcg cccgctaccc cgaccacatg aagcagcacg acttcttcaa
     1201 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaaag acgacggcaa
     1261 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     1321 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
     1381 caacagccac aacgtctata tcaccgccga caagcagaag aacggcatca aggccaactt
     1441 caagatccgc cacaacatcg aggacggcgg cgtgcagctc gccgaccact accagcagaa
     1501 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gctaccagtc
     1561 caagctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
     1621 cgccgccggg atcactctcg gcatggacga gctgtacaag ggctctggtg ctaccaactt
     1681 ctcactgttg aaacaggcag gggatgtaga ggagaatcca gggcctggtg ctagtggaga
     1741 ctacaaagac catgacggag attataaaga tcatgacatc gattacaagg atgacgatga
     1801 caagtccgga ctcagatctg gaggctccgg aggcccagag ccagcgaagt ctgctcccgc
     1861 cccgaaaaag ggctccaaga aggcggtgac taaggcgcag aagaaaggcg gcaagaagcg
     1921 caagcgcagc cgcaaggaga gctattccat ctatgtgtac aaggttctga agcaggtcca
     1981 ccctgacacc ggcatttcgt ccaaggccat gggcatcatg aattcgtttg tgaacgacat
     2041 tttcgagcgc atcgcaggag aggcttcccg cctggcgcat tacaacaagc gctcgaccat
     2101 cacctcccgg gagatccaga cggccgtgcg cctgctgctg cctggggagt tggccaagca
     2161 cgccgtgtcc gagggtacta aggccatcac caagtacacc agcgctaagt aatgaggcgc
     2221 gccgcggccg cgaattcgat atcataatca acctctggat tacaaaattt gtgaaagatt
     2281 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     2341 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     2401 gttagttctt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     2461 tcggctgttg ggcactgaca attccgtggc tcgagagatc ttcgactgtg ccttctagtt
     2521 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     2581 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     2641 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     2701 ggcatgagat ctcacgtgcg gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc
     2761 actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc
     2821 ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc
     2881 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc
     2941 aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     3001 gcgtgaccgc tacacttgcc agcgccttag cgcccgctcc tttcgctttc ttcccttcct
     3061 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     3121 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac
     3181 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     3241 ttaatagtgg actcttgttc caaactggaa caacactcaa ctctatctcg ggctattctt
     3301 ttgatttata agggattttg ccgatttcgg tctattggtt aaaaaatgag ctgatttaac
     3361 aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca
     3421 gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg
     3481 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     3541 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg
     3601 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     3661 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     3721 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     3781 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     3841 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     3901 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     3961 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     4021 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     4081 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     4141 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     4201 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     4261 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     4321 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     4381 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     4441 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     4501 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     4561 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     4621 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     4681 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     4741 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     4801 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     4861 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     4921 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     4981 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     5041 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     5101 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     5161 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     5221 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     5281 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     5341 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     5401 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     5461 ttgctcacat gtcctgcagg cagctgcgcg ctcgctcgct cactgaggcc gcccgggcgt
     5521 cgggcgacct ttggtcgccc ggcctcagtg agcgagcg
//
