LOCUS       sequence-466292-        4745 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2377m 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 4745)
  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 4745)
  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: AiP1700 - pAAV-AiE2377m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4745
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   107..236
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(298..886)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(378..397)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1057..1848,1849..1917))
                     /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     1680..1699
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1918..2022)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2090..2108
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2146..2168)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2268..2287
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2304..2759)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2441..2462)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2653..2672
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2834..2974
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2845..2974
                     /label=AAV2 ITR
     polyA_signal    3092..3203
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3390..3410
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(3503..4213,4214..4216,4217..4219))
                     /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(3545..3566)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3894..3913
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4153..4174
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4216..4225
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 atgagatgtg gtagggttta gctgttaaat ggagctggta acatttagat tcagaggatt
       61 gtactcattg tcttccgatt attactctta caacgcgtgc ggccgcagga acccctagtg
      121 atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag
      181 gtcgcccgac gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggacat
      241 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
      301 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
      361 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
      421 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
      481 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
      541 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
      601 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
      661 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
      721 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
      781 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
      841 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
      901 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
      961 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     1021 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     1081 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     1141 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     1201 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     1261 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     1321 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     1381 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     1441 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     1501 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     1561 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     1621 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     1681 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     1741 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     1801 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     1861 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     1921 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     1981 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     2041 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     2101 cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     2161 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     2221 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
     2281 gattgtactg agagtgcacc ataaaattgt aaacgttaat attttgttaa aattcgcgtt
     2341 aaatttttgt taaatcagct cattttttaa ccaatagacc gaaatcggca aaatccctta
     2401 taaatcaaaa gaatagcccg agatagagtt gagtgttgtt ccagtttgga acaagagtcc
     2461 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     2521 cccactacgt gaaccatcac ccaaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     2581 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     2641 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct aaggcgctgg caagtgtagc
     2701 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgta
     2761 ctatggttgc tttgacgtat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
     2821 cgcatcaggc gcccctgcag gcagctgcgc gctcgctcgc tcactgaggc cgcccgggca
     2881 aagcccgggc gtcgggcgac ctttggtcgc ccggcctcag tgagcgagcg agcgcgcaga
     2941 gagggagtgg ccaactccat cactaggggt tcctgcggcc gctcggtccg cacgtgagat
     3001 ctcatgcctg ctattgtctt cccaatcctc ccccttgctg tcctgcccca ccccaccccc
     3061 cagaatagaa tgacacctac tcagacaatg cgatgcaatt tcctcatttt attaggaaag
     3121 gacagtggga gtggcacctt ccagggtcaa ggaaggcacg ggggaggggc aaacaacaga
     3181 tggctggcaa ctagaaggca cagtcgaaga tctctcgagc cacggaattg tcagtgccca
     3241 acagccgagc ccctgtccag cagcgggcaa ggcaggcggc gatgagttcc gccgtggcaa
     3301 gaactaacca ggatttatac aaggaggaga aaatgaaagc catacgggaa gcaatagcat
     3361 gatacaaagg cattaaagca gcgtatccac atagcgtaaa aggagcaaca tagttaagaa
     3421 taccagtcaa tctttcacaa attttgtaat ccagaggttg attatgatat cgaattcgcg
     3481 gccgcggcgc gccgtcgact tacttgtaca gctcgtccat gccgagagtg atcccggcgg
     3541 cggtcacgaa ctccagcagg accatgtgat cgcgcttctc gttggggtct ttgctcagct
     3601 tggactggta gctcaggtag tggttgtcgg gcagcagcac ggggccgtcg ccgatggggg
     3661 tgttctgctg gtagtggtcg gcgagctgca cgccgccgtc ctcgatgttg tggcggatct
     3721 tgaagttggc cttgatgccg ttcttctgct tgtcggcggt gatatagacg ttgtggctgt
     3781 tgtagttgta ctccagcttg tgccccagga tgttgccgtc ctccttgaag tcgatgccct
     3841 tcagctcgat gcggttcacc agggtgtcgc cctcgaactt cacctcggcg cgggtcttgt
     3901 agttgccgtc gtccttgaag aagatggtgc gctcctggac gtagccttcg ggcatggcgg
     3961 acttgaagaa gtcgtgctgc ttcatgtggt cggggtagcg ggcgaagcac tgcacgccgt
     4021 agcccagggt ggtcacgagg gtgggccagg gcacgggcag cttgccggtg gtgcagatca
     4081 gcttcagggt cagcttgccg taggtggcat cgccctcgcc ctcgccggac acgctgaact
     4141 tgtggccgtt tacgtcgccg tccagctcga ccaggatggg caccaccccg gtgaacagct
     4201 cctcgccctt gctcaccatg gtggcgaccg gtagcgctag cttcgaaaga tctggatccc
     4261 agaagcaaat gtaagcaata gatggctctg ccctgacttt tatgcccagc ccgagctccg
     4321 gtcttcttgc tccacaacag tttctcttag gctccagact taggcttcca aatctctgga
     4381 ttttgctcct tcaatttgct gggctgctat gacatagaaa tatgcccaga aatggtaaaa
     4441 cggtgtataa tttgcttctc aatcttgaac tgattagaat ctcaaatgaa aactatcttg
     4501 taagttttga atgggcctat tattaaatgc tacagatatc agaaagtgtt attactgaat
     4561 tgaagagtca gcccaaggtg aaaacagtaa tgaattccaa gacgtgggtt ggatccctgg
     4621 ctggtcttcc tttagtcaac aaggaaacac tgagctagct agtttttccc catgatgatc
     4681 ctgagtctac atgcttatcc ccaaaagaag ataaaatcat ttctcacaca tgcataagca
     4741 cactg
//
