LOCUS       sequence-470601-        5661 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0790h 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 5661)
  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 5661)
  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: AiP14525 -
            pAAV-AiE0790h-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5661
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(339..1130,1131..1199))
                     /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     962..981
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1200..1304)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1372..1390
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1428..1450)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1550..1569
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(1586..2041)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1723..1744)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1935..1954
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     misc_feature    2110..2137
                     /gene="telL"
                     /label=telL
                     /note="telomeric sequence from coliphage N15"
     misc_feature    2138..2165
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     repeat_region   2172..2312
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2183..2312
                     /label=AAV2 ITR
     polyA_signal    2430..2541
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2728..2748
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2835..3212)
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     CDS             complement(3243..3308)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             complement(3321..3377)
                     /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             complement(join(3387..4097,4098..4100,4101..4103))
                     /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(3429..3450)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3778..3797
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     regulatory      4100..4109
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      4345..4354
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   4993..5122
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     misc_feature    5129..5156
                     /gene="telL"
                     /label=telL
                     /note="telomeric sequence from coliphage N15"
     misc_feature    5157..5184
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     rep_origin      complement(join(5241..5661,1..168))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5321..5340)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
       61 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
      121 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
      181 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
      241 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
      301 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
      361 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
      421 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
      481 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
      541 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
      601 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
      661 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
      721 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
      781 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
      841 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
      901 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
      961 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     1021 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     1081 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     1141 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     1201 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     1261 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     1321 gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt
     1381 atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
     1441 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt
     1501 cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag
     1561 cagattgtac tgagagtgca ccataaaatt gtaaacgtta atattttgtt aaaattcgcg
     1621 ttaaattttt gttaaatcag ctcatttttt aaccaataga ccgaaatcgg caaaatccct
     1681 tataaatcaa aagaatagcc cgagatagag ttgagtgttg ttccagtttg gaacaagagt
     1741 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     1801 ggcccactac gtgaaccatc acccaaatca agttttttgg ggtcgaggtg ccgtaaagca
     1861 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     1921 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctaaggcgct ggcaagtgta
     1981 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     2041 tactatggtt gctttgacgt atgcggtgtg aaataccgca cagatgcgta aggagaaaat
     2101 accgcatcat atcagcacac aatagtccat tatacgcgcg tataatgggc aattgtgtgc
     2161 tgataggcgc ccctgcaggc agctgcgcgc tcgctcgctc actgaggccg cccgggcaaa
     2221 gcccgggcgt cgggcgacct ttggtcgccc ggcctcagtg agcgagcgag cgcgcagaga
     2281 gggagtggcc aactccatca ctaggggttc ctgcggccgc tcggtccgca cgtgagatct
     2341 catgcctgct attgtcttcc caatcctccc ccttgctgtc ctgccccacc ccacccccca
     2401 gaatagaatg acacctactc agacaatgcg atgcaatttc ctcattttat taggaaagga
     2461 cagtgggagt ggcaccttcc agggtcaagg aaggcacggg ggaggggcaa acaacagatg
     2521 gctggcaact agaaggcaca gtcgaagatc tctcgagcca cggaattgtc agtgcccaac
     2581 agccgagccc ctgtccagca gcgggcaagg caggcggcga tgagttccgc cgtggcaaga
     2641 actaaccagg atttatacaa ggaggagaaa atgaaagcca tacgggaagc aatagcatga
     2701 tacaaaggca ttaaagcagc gtatccacat agcgtaaaag gagcaacata gttaagaata
     2761 ccagtcaatc tttcacaaat tttgtaatcc agaggttgat tatgatatcg aattcgcggc
     2821 cgcggcgcgc ctcattactt agcgctggtg tacttggtga tggccttagt accctcggac
     2881 acggcgtgct tggccaactc cccaggcagc agcaggcgca cggccgtctg gatctccctg
     2941 gaggtgatgg tcgagcgctt gttgtaatgc gccaggcggg aagcctcacc tgcgatgcgc
     3001 tcgaaaatgt cgttcacaaa cgagttcatg atgcccatgg ccttggacga aatgccggtg
     3061 tcagggtgga cctgcttcag aaccttatac acatagatgg aatagctctc cttgcggctg
     3121 cgcttgcgct tcttgccgcc tttcttctgc gccttagtca ccgccttctt ggagcccttt
     3181 ttcggggcgg gagcagactt cgctggctct gggcctccgg agcctccaga tctgagtccg
     3241 gacttgtcat cgtcatcctt gtaatcgatg tcatgatctt tataatctcc gtcatggtct
     3301 ttgtagtctc cactagcacc aggccctgga ttctcctcta catcccctgc ctgtttcaac
     3361 agtgagaagt tggtagcacc agagcccttg tacagctcgt ccatgccgag agtgatcccg
     3421 gcggcggtca cgaactccag caggaccatg tgatcgcgct tctcgttggg gtctttgctc
     3481 agcttggact ggtagctcag gtagtggttg tcgggcagca gcacggggcc gtcgccgatg
     3541 ggggtgttct gctggtagtg gtcggcgagc tgcacgccgc cgtcctcgat gttgtggcgg
     3601 atcttgaagt tggccttgat gccgttcttc tgcttgtcgg cggtgatata gacgttgtgg
     3661 ctgttgtagt tgtactccag cttgtgcccc aggatgttgc cgtcctcctt gaagtcgatg
     3721 cccttcagct cgatgcggtt caccagggtg tcgccctcga acttcacctc ggcgcgggtc
     3781 ttgtagttgc cgtcgtcttt gaagaagatg gtgcgctcct ggacgtagcc ttcgggcatg
     3841 gcggacttga agaagtcgtg ctgcttcatg tggtcggggt agcgggcgaa gcactgcacg
     3901 ccgtagccca gggtggtcac gagggtgggc cagggcacgg gcagcttgcc ggtggtgcag
     3961 atcagcttca gggtcagctt gccgtaggtg gcatcgccct cgccctcgcc ggacacgctg
     4021 aacttgtggc cgttgacatc gccgtccagc tcgaccagga tgggcaccac cccggtgaac
     4081 agctcctcgc ccttggagac catggtggcg accggtagcg ctagcttcga aagatctgga
     4141 tcccagaagc aaatgtaagc aatagatggc tctgccctga cttttatgcc cagcccgagc
     4201 tcttaccagc acagcactgc cctctaggaa ctttatccat tctgcaggga aatggaagat
     4261 catggaagaa agcagtgtgt ggatggtgaa gtgcccttca caggcacttt gattttggag
     4321 gcagaactgg gcctgccaga agaagccacc atggtctcat caggccaggc cccctcagac
     4381 accctggagg ttttgtctgt tgctgcttca ctttcaatgt gtgattccca actgcttagc
     4441 tgccttttac agcgatgggc tgaaatgtcc ccttggtctc ttgctccttc ccttattcct
     4501 ccagccccac ccaccaggag cccgcctgcc actaaattgc catgccagca attttctcag
     4561 ccgcttaaaa ggccatcaag acagcatcaa actgcctgtg catgctaaac ccaaacgctt
     4621 atggagcaga gcccagagtg ggatgctaaa ttggattcta tcaaatcaac atcagctgcg
     4681 tgcaagtcgc tgataatgaa aggctcctaa cgatggcgtt catcgctata aagagatgac
     4741 taagagctaa gaatggatgg ctgctggctc ttgctgggat gggaaggagg caccggggag
     4801 aggctgtggt gatggactat cccttgtgga tgctggctgg agaggggagg aaggaggagt
     4861 cagataaggg atgttctcca tttttggaag ggggatggat ataggagaga tagtcatgtt
     4921 ctccattctt tcttacacac actcaaaacc cagaaacaca ttgaagtcat gaaggtggac
     4981 gcgtgcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct
     5041 cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggcggcctc agtgagcgag
     5101 cgagcgcgca gctgcctgca ggacatgtta tcagcacaca atagtccatt atacgcgcgt
     5161 ataatgggca attgtgtgct gatatgagca aaaggccagc aaaaggccag gaaccgtaaa
     5221 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     5281 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     5341 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     5401 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     5461 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     5521 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     5581 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     5641 gagttcttga agtggtggcc t
//
