LOCUS       sequence-461848-        4803 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0762m 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 4803)
  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 4803)
  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: AiP12598 - pAAV-AiE0762m-minBG-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..4803
                     /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      815..824
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(821..823,824..826,827..1537)
                     /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(866..887)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1127..1146)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1474..1495
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1630..1650)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1837..1948
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2066..2206
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2066..2195
                     /label=AAV2 ITR
     rep_origin      2281..2736
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2368..2387)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2578..2599
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2753..2772)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2872..2894
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2932..2950)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3018..3122
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3123..3191,3192..3983)
                     /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(3341..3360)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4154..4742
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4643..4662
                     /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 gcgtactatg gatgaaataa aatgtggaaa ggcccacttg
      181 tatttaagtc taaacattga ggatttaaat cagcttcagc tgacgcctag ggaaaaggcc
      241 agtagaattt taatcactaa ttgagcattt ccagttattt ttaaaatgag tgtcataagc
      301 aatgatagac cattctttat attgcttcaa aagtaaaagt cttcagatcc atatgaagct
      361 acttttaaaa tctaggtaca ttgatgtgga tacaatgctt ccaaagaagt attttaaaca
      421 gaactttata ttactctgtg atttaccaga tgcgcaattc atcaggctgt tctatgatct
      481 tggagcaaga agtcacaggt gctgactaga acacacattt aaaagaagag tctgtgctcc
      541 aatcaccatg tattctacag aagttaaatt aaggcagatg ctacagagac aaacactcat
      601 ctccataata tcaaccagcg caagcctagt aacttctaac gccataagtg cgttagaccc
      661 ctgaggatag gtccgaaaaa agaaaagaaa aacattttcc ttgctttctt ccattttacc
      721 agagctcggg ctgggcataa aagtcagggc agagccatct attgcttaca tttgcttctg
      781 ggatccagat ctttcgaagc tagcgctacc ggtcgccacc atggtgagca agggcgagga
      841 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
      901 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagct
      961 gatctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgggcta
     1021 cggcgtgcag tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     1081 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
     1141 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     1201 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
     1261 cagccacaac gtctatatca ccgccgacaa gcagaagaac ggcatcaagg ccaacttcaa
     1321 gatccgccac aacatcgagg acggcggcgt gcagctcgcc gaccactacc agcagaacac
     1381 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagct accagtccaa
     1441 gctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
     1501 cgccgggatc actctcggca tggacgagct gtacaagtaa gtcgacggcg cgccgcggcc
     1561 gcgaattcga tatcataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     1621 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     1681 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttagttct
     1741 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     1801 gggcactgac aattccgtgg ctcgagagat cttcgactgt gccttctagt tgccagccat
     1861 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     1921 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     1981 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgaga
     2041 tctcacgtgc ggaccgagcg gccgcaggaa cccctagtga tggagttggc cactccctct
     2101 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt
     2161 gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg
     2221 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt
     2281 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     2341 ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     2401 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     2461 gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc
     2521 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     2581 gactcttgtt ccaaactgga acaacactca actctatctc gggctattct tttgatttat
     2641 aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta
     2701 acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct
     2761 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct
     2821 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     2881 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga
     2941 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     3001 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     3061 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     3121 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     3181 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     3241 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     3301 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     3361 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     3421 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     3481 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     3541 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     3601 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     3661 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     3721 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     3781 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     3841 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     3901 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     3961 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     4021 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     4081 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     4141 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     4201 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     4261 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     4321 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     4381 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     4441 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     4501 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     4561 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     4621 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     4681 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     4741 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     4801 tgt
//
