LOCUS       sequence-470458-        4752 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0131h 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 4752)
  AUTHORS   Ben-Simon Y, Hooper M, Narayan S, Daigle T, Dwivedi D, Way SW, Oster
            A, Stafford DA, Mich JK, Taormina MJ, Martinez RA, Opitz-Araya X, 
            Roth JR, Allen S, Ayala A, Bakken TE, Barcelli T, Barta S, Bendrick 
            J, Bertagnolli D, Bowlus J, Boyer G, Brouner K, Casian B, Casper T, 
            Chakka AB, Chakrabarty R, Chance RK, Chavan S, Departee M, Donadio 
            N, Dotson N, Egdorf T, Gabitto M, Garcia J, Gary A, Gasperini M, 
            Goldy J, Gore BB, Graybuck L, Greisman N, Haeseleer F, Halterman C, 
            Helback O, Hockemeyer D, Huang C, Huff S, Hunker A, Johansen N, 
            Juneau Z, Kalmbach B, Khem S, Kussick E, Kutsal R, Larsen R, Lee C, 
            Lee AY, Leibly M, Lenz GH, Liang E, Lusk N, Malone J, Mollenkopf T, 
            Morin E, Newman D, Ng L, Ngo K, Omstead V, Oyama A, Pham T, Pom CA, 
            Potekhina L, Ransford S, Rette D, Rimorin C, Rocha D, Ruiz A, 
            Sanchez REA, Sedeno-Cortes A, Sevigny JP, Shapovalova N, Shulga L, 
            Sigler AR, Siverts LA, Somasundaram S, Stewart K, Szelenyi E, Tieu 
            M, Trader C, van Velthoven CTJ, Walker M, 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 J, Tasic B
  TITLE     A suite of enhancer AAVs and transgenic mouse lines for genetic 
            access to cortical cell types (preprint).
  JOURNAL   bioRxiv [Preprint]. 2024 Sep 26:2024.06.10.597244. doi: 
            10.1101/2024.06.10.597244.
  PUBMED    38915722
REFERENCE   2  (bases 1 to 4752)
  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: "bioRxiv 
            [Preprint]. 2024 Sep 26:2024.06.10.597244. doi: 
            10.1101/2024.06.10.597244."
COMMENT     Sequence Label: AiP12310 - pAAV-AiE0131h-minBG-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..4752
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      270..279
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(276..278,279..281,282..992)
                     /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(321..342)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(582..601)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     929..950
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1085..1105)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1292..1403
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1521..1661
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1521..1650
                     /label=AAV2 ITR
     rep_origin      1736..2191
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1823..1842)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2033..2054
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2208..2227)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2327..2349
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2387..2405)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2473..2577
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2578..2646,2647..3438)
                     /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(2796..2815)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3609..4197
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4098..4117
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   4259..4388
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 tgaaacttct gacttaaagc tctgaaagaa atgccactat atatattttt ctagcccata
       61 gaaggataaa ctttttgccc tctgattgca gtggggatga ggagtccaag taacatgata
      121 ttcatattgt taaaactgat ccctcgcatt gcctctttag ggtcagtgac tgctaggagc
      181 tcgggctggg cataaaagtc agggcagagc catctattgc ttacatttgc ttctgggatc
      241 cagatctttc gaagctagcg ctaccggtcg ccaccatggt gagcaagggc gaggagctgt
      301 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
      361 gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagctgatct
      421 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg ggctacggcg
      481 tgcagtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
      541 tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga
      601 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
      661 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc
      721 acaacgtcta tatcaccgcc gacaagcaga agaacggcat caaggccaac ttcaagatcc
      781 gccacaacat cgaggacggc ggcgtgcagc tcgccgacca ctaccagcag aacaccccca
      841 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagctaccag tccaagctga
      901 gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg
      961 ggatcactct cggcatggac gagctgtaca agtaagtcga cggcgcgccg cggccgcgaa
     1021 ttcgatatca taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     1081 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     1141 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggtta gttcttgcca
     1201 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca
     1261 ctgacaattc cgtggctcga gagatcttcg actgtgcctt ctagttgcca gccatctgtt
     1321 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     1381 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     1441 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgagatctca
     1501 cgtgcggacc gagcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg
     1561 cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg
     1621 ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt
     1681 tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg
     1741 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     1801 cttgccagcg ccttagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     1861 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     1921 ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg
     1981 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     2041 ttgttccaaa ctggaacaac actcaactct atctcgggct attcttttga tttataaggg
     2101 attttgccga tttcggtcta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     2161 aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct
     2221 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     2281 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     2341 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
     2401 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     2461 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     2521 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     2581 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     2641 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     2701 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     2761 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     2821 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     2881 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     2941 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     3001 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     3061 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     3121 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     3181 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     3241 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     3301 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     3361 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     3421 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     3481 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     3541 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     3601 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     3661 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     3721 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     3781 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     3841 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     3901 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     3961 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     4021 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     4081 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     4141 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     4201 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgtcc
     4261 tgcaggcagc tgcgcgctcg ctcgctcact gaggccgccc gggcgtcggg cgacctttgg
     4321 tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga gtggccaact ccatcactag
     4381 gggttcctgc ggccgcacgc gttattttgg gatgtataaa tcagacgaaa agtcacaaaa
     4441 tcgtgtgcca ttcaaaggtg gtggaagttg aagaggaagc gtatctaaat gacagataat
     4501 gagtctttat gaagttacga aaaagaggtc agcacctgag aaacagacta aactagaata
     4561 gtcatatgac atctgatgtt cactgtaaaa tgagatgagt gctgttggtg taatgaggtt
     4621 ccttagtatg ctactaagtt taactgcata aattatacca attagctcca aaatgaatgt
     4681 aaatcctcaa ctataaaatg tttcaatgtg tacttagtgc taataggaaa tgtttgtggc
     4741 tgtgagccat cc
//
