LOCUS       sequence-470515-        4841 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0708m 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 4841)
  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 4841)
  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: AiP12715 - pAAV-AiE0708m-minBG-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..4841
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      315..324
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(321..323,324..326,327..1037)
                     /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(366..387)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(627..646)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     974..995
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1130..1150)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1337..1448
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1566..1706
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1566..1695
                     /label=AAV2 ITR
     rep_origin      1781..2236
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1868..1887)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2078..2099
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2253..2272)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2372..2394
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2432..2450)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2518..2622
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2623..2691,2692..3483)
                     /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(2841..2860)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3654..4242
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4143..4162
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   4304..4433
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 ctaacagctg ctggaaggtc agggtggatg cagacttgtg gcatcctctg ctttccctgg
       61 caaagcacta atcaaacatt gctagaattg tcttcaaata gagcttgaga gtctgaaaca
      121 ggtctgtcca tgtgtggagc atcaagccag acacatcagc aaaccactat ggaaaagcct
      181 tcatggcaaa aaaatggcaa gggccaactg tgcaggctgt ggagctcggg ctgggcataa
      241 aagtcagggc agagccatct attgcttaca tttgcttctg ggatccagat ctttcgaagc
      301 tagcgctacc ggtcgccacc atggtgagca agggcgagga gctgttcacc ggggtggtgc
      361 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
      421 gcgagggcga tgccacctac ggcaagctga ccctgaagct gatctgcacc accggcaagc
      481 tgcccgtgcc ctggcccacc ctcgtgacca ccctgggcta cggcgtgcag tgcttcgccc
      541 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
      601 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
      661 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
      721 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
      781 ccgccgacaa gcagaagaac ggcatcaagg ccaacttcaa gatccgccac aacatcgagg
      841 acggcggcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
      901 tgctgctgcc cgacaaccac tacctgagct accagtccaa gctgagcaaa gaccccaacg
      961 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
     1021 tggacgagct gtacaagtaa gtcgacggcg cgccgcggcc gcgaattcga tatcataatc
     1081 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
     1141 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
     1201 ctttcatttt ctcctccttg tataaatcct ggttagttct tgccacggcg gaactcatcg
     1261 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg
     1321 ctcgagagat cttcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     1381 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     1441 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     1501 agcaaggggg aggattggga agacaatagc aggcatgaga tctcacgtgc ggaccgagcg
     1561 gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac
     1621 tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag
     1681 cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc ttacgcatct
     1741 gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca
     1801 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta
     1861 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     1921 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     1981 cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     2041 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     2101 acaacactca actctatctc gggctattct tttgatttat aagggatttt gccgatttcg
     2161 gtctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     2221 ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     2281 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     2341 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     2401 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt
     2461 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     2521 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     2581 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     2641 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     2701 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     2761 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     2821 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     2881 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     2941 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     3001 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     3061 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     3121 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     3181 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     3241 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     3301 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     3361 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     3421 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     3481 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     3541 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     3601 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     3661 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     3721 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     3781 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     3841 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     3901 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     3961 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     4021 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     4081 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     4141 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     4201 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     4261 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgtcctgcag gcagctgcgc
     4321 gctcgctcgc tcactgaggc cgcccgggcg tcgggcgacc tttggtcgcc cggcctcagt
     4381 gagcgagcga gcgcgcagag agggagtggc caactccatc actaggggtt cctgcggccg
     4441 cacgcgttct ccagcatcac tttcacccac tgttgcagct gggaggcacc ttctgactgt
     4501 gcctgctcac cctcgaacca ggaagccaaa ggccacccac aggctacagt acacaggtca
     4561 cctcatgcca ctgaagggac ccgagaggga gagagcctgg ttgaggtggc agctgcttaa
     4621 tgacctagat gtcagagctg atgggctact actgtctgct catcttgagc agagaaactg
     4681 ggtcagccaa gacaagacac caagagccgt gtttggcaaa acaccattag ttccgtggaa
     4741 caaatcttga tagactcatt tagtgtctcc cgtctattaa tttaatggcc aataaaagca
     4801 atggctgggg agaaagctca gaacaagggc aaagctagtc g
//
