LOCUS       sequence-461880-        5006 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0980h 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 5006)
  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 5006)
  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: AiP13488 -
            pAAV-AiE0980h-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5006
                     /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      472..481
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(478..480,481..483,484..1194)
                     /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(784..803)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1131..1152
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1204..1260
                     /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             1273..1338
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1369..1746
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(1833..1853)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2040..2151
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2269..2409
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2269..2398
                     /label=AAV2 ITR
     rep_origin      2484..2939
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2571..2590)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2781..2802
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2956..2975)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3075..3097
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3135..3153)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3221..3325
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3326..3394,3395..4186)
                     /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(3544..3563)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4357..4945
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4846..4865
                     /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 gcgtattgca ggagacattt tccccggctc ccactttctt
      181 cccactaacc acccccacca cgcaggtgct tcttatgtgg gccctttcca cagtcttgga
      241 atatctgctc actccaccag gtgctgctct cactcccagc accaccctcc cctcaccccc
      301 acattccatt ccctggagtg gctgcaagaa tcttttccag gagcataagc ctttatctca
      361 ggtgagaaga ctgcttgaga gctcgggctg ggcataaaag tcagggcaga gccatctatt
      421 gcttacattt gcttctggga tccagatctt tcgaagctag cgctaccggt cgccaccatg
      481 gtctccaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
      541 gatgtcaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
      601 aagctgaccc tgaagctgat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
      661 gtgaccaccc tgggctacgg cgtgcagtgc ttcgcccgct accccgacca catgaagcag
      721 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
      781 aaagacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
      841 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
      901 ctggagtaca actacaacag ccacaacgtc tatatcaccg ccgacaagca gaagaacggc
      961 atcaaggcca acttcaagat ccgccacaac atcgaggacg gcggcgtgca gctcgccgac
     1021 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     1081 ctgagctacc agtccaagct gagcaaagac cccaacgaga agcgcgatca catggtcctg
     1141 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagggctct
     1201 ggtgctacca acttctcact gttgaaacag gcaggggatg tagaggagaa tccagggcct
     1261 ggtgctagtg gagactacaa agaccatgac ggagattata aagatcatga catcgattac
     1321 aaggatgacg atgacaagtc cggactcaga tctggaggct ccggaggccc agagccagcg
     1381 aagtctgctc ccgccccgaa aaagggctcc aagaaggcgg tgactaaggc gcagaagaaa
     1441 ggcggcaaga agcgcaagcg cagccgcaag gagagctatt ccatctatgt gtacaaggtt
     1501 ctgaagcagg tccaccctga caccggcatt tcgtccaagg ccatgggcat catgaattcg
     1561 tttgtgaacg acattttcga gcgcatcgca ggagaggctt cccgcctggc gcattacaac
     1621 aagcgctcga ccatcacctc ccgggagatc cagacggccg tgcgcctgct gctgcctggg
     1681 gagttggcca agcacgccgt gtccgagggt actaaggcca tcaccaagta caccagcgct
     1741 aagtaatgag gcgcgccgcg gccgcgaatt cgatatcata atcaacctct ggattacaaa
     1801 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     1861 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     1921 ttgtataaat cctggttagt tcttgccacg gcggaactca tcgccgcctg ccttgcccgc
     1981 tgctggacag gggctcggct gttgggcact gacaattccg tggctcgaga gatcttcgac
     2041 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     2101 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     2161 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     2221 ggaagacaat agcaggcatg agatctcacg tgcggaccga gcggccgcag gaacccctag
     2281 tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa
     2341 aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct
     2401 gcctgcaggg gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     2461 gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     2521 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc
     2581 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     2641 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt
     2701 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     2761 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaactctat
     2821 ctcgggctat tcttttgatt tataagggat tttgccgatt tcggtctatt ggttaaaaaa
     2881 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt
     2941 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     3001 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     3061 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     3121 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     3181 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     3241 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     3301 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     3361 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     3421 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     3481 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     3541 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     3601 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     3661 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     3721 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     3781 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     3841 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     3901 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     3961 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     4021 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     4081 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     4141 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     4201 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     4261 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     4321 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     4381 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     4441 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     4501 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     4561 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     4621 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     4681 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     4741 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     4801 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     4861 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     4921 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     4981 cttttgctgg ccttttgctc acatgt
//
