LOCUS       sequence-470507-        4790 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0713h 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 4790)
  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 4790)
  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: AiP12693 - pAAV-AiE0713h-minBG-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..4790
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      289..298
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(295..297,298..300,301..1011)
                     /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(340..361)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(601..620)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     948..969
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1104..1124)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1311..1422
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1540..1680
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1540..1669
                     /label=AAV2 ITR
     rep_origin      1755..2210
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1842..1861)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2052..2073
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2227..2246)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2346..2368
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2406..2424)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2492..2596
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2597..2665,2666..3457)
                     /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(2815..2834)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3628..4216
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4117..4136
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   4278..4407
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 tagaagtttg cttaatttct catttttgtc tgcagtgttt gcttttgttt gtctgtttga
       61 agtaatgcct gatgacagct taatatatgc aagttggctt ccagtagctt ctcttggttc
      121 tgtgttacca gttaagcctt gaggacctcc tgtgtgtcag gggtttcctg gatacagaaa
      181 ggcacagaga aagccgagct cgggctgggc ataaaagtca gggcagagcc atctattgct
      241 tacatttgct tctgggatcc agatctttcg aagctagcgc taccggtcgc caccatggtg
      301 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
      361 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
      421 ctgaccctga agctgatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
      481 accaccctgg gctacggcgt gcagtgcttc gcccgctacc ccgaccacat gaagcagcac
      541 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
      601 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
      661 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
      721 gagtacaact acaacagcca caacgtctat atcaccgccg acaagcagaa gaacggcatc
      781 aaggccaact tcaagatccg ccacaacatc gaggacggcg gcgtgcagct cgccgaccac
      841 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
      901 agctaccagt ccaagctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
      961 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaagtcgac
     1021 ggcgcgccgc ggccgcgaat tcgatatcat aatcaacctc tggattacaa aatttgtgaa
     1081 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     1141 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     1201 tcctggttag ttcttgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     1261 ggggctcggc tgttgggcac tgacaattcc gtggctcgag agatcttcga ctgtgccttc
     1321 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     1381 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     1441 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     1501 tagcaggcat gagatctcac gtgcggaccg agcggccgca ggaaccccta gtgatggagt
     1561 tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc
     1621 gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg
     1681 ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgtc
     1741 aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     1801 gcgcagcgtg accgctacac ttgccagcgc cttagcgccc gctcctttcg ctttcttccc
     1861 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     1921 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg
     1981 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     2041 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaactcta tctcgggcta
     2101 ttcttttgat ttataaggga ttttgccgat ttcggtctat tggttaaaaa atgagctgat
     2161 ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt tatggtgcac
     2221 tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc
     2281 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     2341 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg
     2401 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     2461 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     2521 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     2581 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     2641 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     2701 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     2761 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     2821 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     2881 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     2941 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     3001 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     3061 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     3121 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     3181 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     3241 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     3301 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     3361 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     3421 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     3481 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     3541 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     3601 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     3661 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     3721 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     3781 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     3841 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     3901 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     3961 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     4021 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     4081 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     4141 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     4201 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     4261 gccttttgct cacatgtcct gcaggcagct gcgcgctcgc tcgctcactg aggccgcccg
     4321 ggcgtcgggc gacctttggt cgcccggcct cagtgagcga gcgagcgcgc agagagggag
     4381 tggccaactc catcactagg ggttcctgcg gccgcacgcg tccttctctc tttgcactca
     4441 gcttcttcag atataagaca ggtgtagtgg gaataacatg cactgtcatt gtgggatgca
     4501 aaaacagaaa actacatgga gtgccctttg tacaccatca ggatttgcac agacatggat
     4561 ttttacagat acctgtgtca gtcataactc tcagtgtccc aggtggtaat cacctaacat
     4621 ctgtgcgggc acttagtagt agagctattg cttttttatt ttccccaagg aagcattgag
     4681 acaagtgctt gaagctagca caagagaggg ctctcaatgg aaatggtgta ggatggtgtg
     4741 cataacagca gcaagtggca aaattatagc tcaatccagt tgcattcata
//
