LOCUS       sequence-461851-        5127 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0719h 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 5127)
  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 5127)
  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: AiP12652 - pAAV-AiE0719h-minBG-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..5127
                     /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      1139..1148
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1145..1147,1148..1150,1151..1861)
                     /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(1190..1211)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1451..1470)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1798..1819
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1954..1974)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2161..2272
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2390..2530
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2390..2519
                     /label=AAV2 ITR
     rep_origin      2605..3060
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2692..2711)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2902..2923
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3077..3096)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3196..3218
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3256..3274)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3342..3446
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3447..3515,3516..4307)
                     /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(3665..3684)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4478..5066
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4967..4986
                     /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 gcgttccatc attccaccca taaaccaatc aacatttatg
      181 agatgtctcc tgtgaatcac atactgtgcc actttttgaa ataaaatgta gagaagttac
      241 aggcctttgt agaagagaaa gtggcataaa aggataattt taagatatgt gacaaattta
      301 agccagtttt gaggactact gctatagata taatttattt aagatggagg ttttctggca
      361 aggtgggatg tgattctatg ggaatctggg ctgctggtga tagttcttgt gaacaactgt
      421 gttgtccaga ttgggctgcg ggtggaaggg tgggtgggga gtgtgggaag gaaggagagg
      481 aagagagagg gagagagtac aagcctaata gactgggaga aatgtagcta tcagaggagt
      541 agaggtctta atatgtgtca gggcggagag tgagaaagct ggaaagatag gaaatgggga
      601 ttggtggttg tgattgtggt gatggggggg gaattttaga tgtcaaagat gaagacaggg
      661 tttaatgtgt ggatagggga gacgatggaa gagctggagg ggaagtggaa gtctcagcat
      721 tgaggatgta gaataattat tatgcagggg tgctgagtag gctctgtgaa tgtggtgaga
      781 gttgccttag acttgggatg tcatgaagac cacaggccag gtatcagtga gtatgtggaa
      841 gtgactaggc gatctttata aaacaagaca ggcgtaaaat ctggaagtag cccaaggagt
      901 aacatattca catctgagac agagattgtg agttgacttt ccatatctgc tcttccttaa
      961 aaatataagc ataattttta gctgagcaca tggccaccca gagcaagact acatttctca
     1021 gaccaccttg caactagtgt gaccagagct cgggctgggc ataaaagtca gggcagagcc
     1081 atctattgct tacatttgct tctgggatcc agatctttcg aagctagcgc taccggtcgc
     1141 caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     1201 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
     1261 ctacggcaag ctgaccctga agctgatctg caccaccggc aagctgcccg tgccctggcc
     1321 caccctcgtg accaccctgg gctacggcgt gcagtgcttc gcccgctacc ccgaccacat
     1381 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
     1441 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     1501 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     1561 gcacaagctg gagtacaact acaacagcca caacgtctat atcaccgccg acaagcagaa
     1621 gaacggcatc aaggccaact tcaagatccg ccacaacatc gaggacggcg gcgtgcagct
     1681 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     1741 ccactacctg agctaccagt ccaagctgag caaagacccc aacgagaagc gcgatcacat
     1801 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     1861 gtaagtcgac ggcgcgccgc ggccgcgaat tcgatatcat aatcaacctc tggattacaa
     1921 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     1981 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     2041 cttgtataaa tcctggttag ttcttgccac ggcggaactc atcgccgcct gccttgcccg
     2101 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggctcgag agatcttcga
     2161 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     2221 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     2281 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     2341 gggaagacaa tagcaggcat gagatctcac gtgcggaccg agcggccgca ggaaccccta
     2401 gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca
     2461 aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc
     2521 tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     2581 cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     2641 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cttagcgccc gctcctttcg
     2701 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     2761 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     2821 tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     2881 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaactcta
     2941 tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggtctat tggttaaaaa
     3001 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt
     3061 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc
     3121 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
     3181 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
     3241 gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
     3301 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     3361 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     3421 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     3481 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     3541 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     3601 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     3661 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     3721 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     3781 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     3841 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     3901 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     3961 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     4021 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     4081 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     4141 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     4201 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     4261 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     4321 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     4381 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     4441 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     4501 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     4561 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     4621 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     4681 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     4741 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     4801 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     4861 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     4921 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     4981 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     5041 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     5101 ccttttgctg gccttttgct cacatgt
//
