LOCUS       sequence-463078-        4753 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE3308m 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 4753)
  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 4753)
  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: AiP20153 - pAAV-AiE3308m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4753
                     /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      765..774
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(771..773,774..776,777..1487)
                     /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(816..837)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1077..1096)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1424..1445
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1580..1600)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1787..1898
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2016..2156
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2016..2145
                     /label=AAV2 ITR
     rep_origin      2231..2686
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2318..2337)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2528..2549
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2703..2722)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2822..2844
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2882..2900)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2968..3072
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3073..3141,3142..3933)
                     /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(3291..3310)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4104..4692
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4593..4612
                     /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 gcgtactgtc tttcatttcc tgtaacatat tgagatattt
      181 ggtgccagcc catttctccc catgccagga aggctggtgt gaaggacctt ttaggtgcat
      241 tgttttctga gggagaattc acagccagtc ttgaagacat gcttaggcaa catctgctaa
      301 tagaagttct agaattatct taacatctaa gaagactcac tgaacaaaaa ttagatttcc
      361 tatatgccaa cgcgcctcgt caactgaaag ctggaagcta aatttggccg taagaaactg
      421 gcttcgaact ccagaagttc tggggccaaa ttcagcctcg ttttacttgt attccttgtt
      481 tcatagggaa aaaaaaatag gtctgctcca gtgacagaga gtctcccatt tattcatctc
      541 aaagtaataa gatcccagaa tggagccttc atgacacaaa gtgcttttaa gttctccctg
      601 aggggagcac aatatacaca atttatgtaa aaagacgaaa aaatgaaagc tgagtattct
      661 aggcgccggc ggagctcggg ctgggcataa aagtcagggc agagccatct attgcttaca
      721 tttgcttctg ggatccagat ctttcgaagc tagcgctacc ggtcgccacc atggtgagca
      781 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
      841 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
      901 ccctgaagct gatctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
      961 ccctgggcta cggcgtgcag tgcttcgccc gctaccccga ccacatgaag cagcacgact
     1021 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
     1081 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     1141 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     1201 acaactacaa cagccacaac gtctatatca ccgccgacaa gcagaagaac ggcatcaagg
     1261 ccaacttcaa gatccgccac aacatcgagg acggcggcgt gcagctcgcc gaccactacc
     1321 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagct
     1381 accagtccaa gctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     1441 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa gtcgacggcg
     1501 cgccgcggcc gcgaattcga tatcataatc aacctctgga ttacaaaatt tgtgaaagat
     1561 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     1621 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     1681 ggttagttct tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
     1741 ctcggctgtt gggcactgac aattccgtgg ctcgagagat cttcgactgt gccttctagt
     1801 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     1861 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     1921 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     1981 aggcatgaga tctcacgtgc ggaccgagcg gccgcaggaa cccctagtga tggagttggc
     2041 cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg
     2101 cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg
     2161 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag
     2221 caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     2281 agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc
     2341 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     2401 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca
     2461 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     2521 tttaatagtg gactcttgtt ccaaactgga acaacactca actctatctc gggctattct
     2581 tttgatttat aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa
     2641 caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc
     2701 agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct
     2761 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     2821 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag
     2881 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     2941 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     3001 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     3061 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     3121 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     3181 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     3241 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     3301 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     3361 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     3421 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     3481 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     3541 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     3601 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     3661 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     3721 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     3781 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     3841 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     3901 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     3961 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     4021 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     4081 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     4141 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     4201 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     4261 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     4321 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     4381 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     4441 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     4501 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     4561 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     4621 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     4681 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     4741 tttgctcaca tgt
//
