LOCUS       sequence-470670-        5135 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0978h 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 5135)
  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 5135)
  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: AiP13462 -
            pAAV-AiE0978h-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5135
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      587..1175
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1076..1095
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   1237..1366
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      1837..1846
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1843..1845,1846..1848,1849..2559)
                     /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(2149..2168)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2496..2517
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2569..2625
                     /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             2638..2703
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2734..3111
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(3198..3218)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    3405..3516
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3634..3774
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3634..3763
                     /label=AAV2 ITR
     rep_origin      3849..4304
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3936..3955)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4146..4167
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4321..4340)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4440..4462
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4500..4518)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4586..4690
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4691..4759,4760..5135,1..416)
                     /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(4909..4928)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
       61 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
      121 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
      181 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
      241 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
      301 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
      361 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
      421 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
      481 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
      541 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
      601 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
      661 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
      721 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
      781 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
      841 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
      901 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
      961 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     1021 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     1081 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     1141 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     1201 ggttcctggc cttttgctgg ccttttgctc acatgtcctg caggcagctg cgcgctcgct
     1261 cgctcactga ggccgcccgg gcgtcgggcg acctttggtc gcccggcctc agtgagcgag
     1321 cgagcgcgca gagagggagt ggccaactcc atcactaggg gttcctgcgg ccgcacgcgt
     1381 tgtgtgcctt gtgaggaatc caggagatct ccaacccgac accaatgcca ggaggggcct
     1441 ggtaaaggcc ttcatgcaat gcagacctcc ttgctctgtg tttcttccca aggcctgtgt
     1501 gcaatgcaga ctttctccct ctatgctcta attttcttcc tggtttcctc tcagcaactc
     1561 gcacctccac tcccacaatc tgttggaaat tttcctgttc cctccgttcc tctctctttt
     1621 gctcaccttt cttgcttttc tcccgccaca gccctacaca acgcacacct ctccctccca
     1681 caatccagag caatttgtgt agcggccacc ccatgtaata tattgaatct gggcacaagc
     1741 agggagctcg ggctgggcat aaaagtcagg gcagagccat ctattgctta catttgcttc
     1801 tgggatccag atctttcgaa gctagcgcta ccggtcgcca ccatggtctc caagggcgag
     1861 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgatgt caacggccac
     1921 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
     1981 ctgatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgggc
     2041 tacggcgtgc agtgcttcgc ccgctacccc gaccacatga agcagcacga cttcttcaag
     2101 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaaaga cgacggcaac
     2161 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
     2221 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
     2281 aacagccaca acgtctatat caccgccgac aagcagaaga acggcatcaa ggccaacttc
     2341 aagatccgcc acaacatcga ggacggcggc gtgcagctcg ccgaccacta ccagcagaac
     2401 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag ctaccagtcc
     2461 aagctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     2521 gccgccggga tcactctcgg catggacgag ctgtacaagg gctctggtgc taccaacttc
     2581 tcactgttga aacaggcagg ggatgtagag gagaatccag ggcctggtgc tagtggagac
     2641 tacaaagacc atgacggaga ttataaagat catgacatcg attacaagga tgacgatgac
     2701 aagtccggac tcagatctgg aggctccgga ggcccagagc cagcgaagtc tgctcccgcc
     2761 ccgaaaaagg gctccaagaa ggcggtgact aaggcgcaga agaaaggcgg caagaagcgc
     2821 aagcgcagcc gcaaggagag ctattccatc tatgtgtaca aggttctgaa gcaggtccac
     2881 cctgacaccg gcatttcgtc caaggccatg ggcatcatga attcgtttgt gaacgacatt
     2941 ttcgagcgca tcgcaggaga ggcttcccgc ctggcgcatt acaacaagcg ctcgaccatc
     3001 acctcccggg agatccagac ggccgtgcgc ctgctgctgc ctggggagtt ggccaagcac
     3061 gccgtgtccg agggtactaa ggccatcacc aagtacacca gcgctaagta atgaggcgcg
     3121 ccgcggccgc gaattcgata tcataatcaa cctctggatt acaaaatttg tgaaagattg
     3181 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     3241 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     3301 ttagttcttg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
     3361 cggctgttgg gcactgacaa ttccgtggct cgagagatct tcgactgtgc cttctagttg
     3421 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     3481 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     3541 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
     3601 gcatgagatc tcacgtgcgg accgagcggc cgcaggaacc cctagtgatg gagttggcca
     3661 ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc
     3721 cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc
     3781 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata cgtcaaagca
     3841 accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     3901 cgtgaccgct acacttgcca gcgccttagc gcccgctcct ttcgctttct tcccttcctt
     3961 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     4021 ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg
     4081 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     4141 taatagtgga ctcttgttcc aaactggaac aacactcaac tctatctcgg gctattcttt
     4201 tgatttataa gggattttgc cgatttcggt ctattggtta aaaaatgagc tgatttaaca
     4261 aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt gcactctcag
     4321 tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga
     4381 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     4441 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg
     4501 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     4561 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     4621 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     4681 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     4741 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     4801 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     4861 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     4921 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     4981 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     5041 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     5101 gacaacgatc ggaggaccga aggagctaac cgctt
//
