LOCUS       sequence-438956-        6145 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2190m 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 6145)
  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 6145)
  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: AiP1511 - pAAV-AiE2190m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..6145
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1127..1256
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(1318..1906)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1398..1417)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2077..2868,2869..2937))
                     /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     2700..2719
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2938..3042)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3110..3128
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3166..3188)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3288..3307
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(3324..3779)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3461..3482)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3673..3692
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   3854..3994
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3865..3994
                     /label=AAV2 ITR
     polyA_signal    4112..4223
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     4410..4430
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(4523..5233,5234..5236,5237..5239))
                     /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(4565..4586)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4914..4933
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5173..5194
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      5236..5245
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 tgaggaactg gtgctttctc ctgtggattc agggaactga acgctgtctg tcaggtttgc
       61 atagcaagtg cttctacctg ctgagacatc tcacctactc agaattcaca cttagaaatg
      121 aagctgtctg ctatgctcta cagcatgtaa gcattcaagt tgtaaaccac tggtagaatc
      181 atcctggcat catagatgga tttaaagtaa cagacacaaa gtgtgtgtgt gtgtgtgttg
      241 gggggggggt tgggggtgcg ctcccttcac ttagattgag gattctcaga attttccagt
      301 gactgacaag tctacagctc caaatcaggt ctcaaggctc ccagtttatc ttctcagtta
      361 caattaagga tgagaataga aatgtcttgg ccctgcagca agggaagata ccagaaggga
      421 acagtgaaaa gatgctagaa gctggagact tgagctggtg acaggctgat taaaggtact
      481 tgaatgttgt atgaagatta attctgcctg cagacctatt ttgcaagtgt ggtaggtatt
      541 gtagggggtg tgctggagct agtagagata gaggaagcct tgggctggct aacttgagtg
      601 actggagacc tcaaggttta tgacatagag atgaggtaga aactggcacc ttgtgccttg
      661 atgaggttac ttgacttgcg ccttttcttt ctgcatgcca aattgagagg ttactggtag
      721 ttttggggtt gggacttgct tctttgaaaa gcaggaaagt ccttaatgtc ccagatcata
      781 tggccttctc cacatccctc gtgcttgttg gccacccagc ctgcctgtat acacaaccta
      841 tttctcccaa aacaatgcca tgacttgagt cacacaagcc ctcagtgaag ggctgagaag
      901 tccttgatag cacctcttct gagcttactt cagtgtaagc ctcttcagtc ctgtactctg
      961 gggaccgagg ggaaatcctg gaccctccca actctgcttc tacttagttt tccaaacaca
     1021 tagctgtgcc atatcttcac gaggagccgt gttagagaag caaagcatac attcctgccc
     1081 agattaatcc gccggcgatt attactctta caacgcgtgc ggccgcagga acccctagtg
     1141 atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag
     1201 gtcgcccgac gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggacat
     1261 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     1321 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     1381 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     1441 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     1501 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     1561 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     1621 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     1681 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     1741 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     1801 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     1861 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     1921 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     1981 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     2041 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     2101 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     2161 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     2221 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     2281 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     2341 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     2401 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     2461 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     2521 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     2581 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     2641 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     2701 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     2761 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     2821 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     2881 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     2941 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     3001 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     3061 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     3121 cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     3181 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     3241 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
     3301 gattgtactg agagtgcacc ataaaattgt aaacgttaat attttgttaa aattcgcgtt
     3361 aaatttttgt taaatcagct cattttttaa ccaatagacc gaaatcggca aaatccctta
     3421 taaatcaaaa gaatagcccg agatagagtt gagtgttgtt ccagtttgga acaagagtcc
     3481 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     3541 cccactacgt gaaccatcac ccaaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     3601 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     3661 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct aaggcgctgg caagtgtagc
     3721 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgta
     3781 ctatggttgc tttgacgtat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
     3841 cgcatcaggc gcccctgcag gcagctgcgc gctcgctcgc tcactgaggc cgcccgggca
     3901 aagcccgggc gtcgggcgac ctttggtcgc ccggcctcag tgagcgagcg agcgcgcaga
     3961 gagggagtgg ccaactccat cactaggggt tcctgcggcc gctcggtccg cacgtgagat
     4021 ctcatgcctg ctattgtctt cccaatcctc ccccttgctg tcctgcccca ccccaccccc
     4081 cagaatagaa tgacacctac tcagacaatg cgatgcaatt tcctcatttt attaggaaag
     4141 gacagtggga gtggcacctt ccagggtcaa ggaaggcacg ggggaggggc aaacaacaga
     4201 tggctggcaa ctagaaggca cagtcgaaga tctctcgagc cacggaattg tcagtgccca
     4261 acagccgagc ccctgtccag cagcgggcaa ggcaggcggc gatgagttcc gccgtggcaa
     4321 gaactaacca ggatttatac aaggaggaga aaatgaaagc catacgggaa gcaatagcat
     4381 gatacaaagg cattaaagca gcgtatccac atagcgtaaa aggagcaaca tagttaagaa
     4441 taccagtcaa tctttcacaa attttgtaat ccagaggttg attatgatat cgaattcgcg
     4501 gccgcggcgc gccgtcgact tacttgtaca gctcgtccat gccgagagtg atcccggcgg
     4561 cggtcacgaa ctccagcagg accatgtgat cgcgcttctc gttggggtct ttgctcagct
     4621 tggactggta gctcaggtag tggttgtcgg gcagcagcac ggggccgtcg ccgatggggg
     4681 tgttctgctg gtagtggtcg gcgagctgca cgccgccgtc ctcgatgttg tggcggatct
     4741 tgaagttggc cttgatgccg ttcttctgct tgtcggcggt gatatagacg ttgtggctgt
     4801 tgtagttgta ctccagcttg tgccccagga tgttgccgtc ctccttgaag tcgatgccct
     4861 tcagctcgat gcggttcacc agggtgtcgc cctcgaactt cacctcggcg cgggtcttgt
     4921 agttgccgtc gtccttgaag aagatggtgc gctcctggac gtagccttcg ggcatggcgg
     4981 acttgaagaa gtcgtgctgc ttcatgtggt cggggtagcg ggcgaagcac tgcacgccgt
     5041 agcccagggt ggtcacgagg gtgggccagg gcacgggcag cttgccggtg gtgcagatca
     5101 gcttcagggt cagcttgccg taggtggcat cgccctcgcc ctcgccggac acgctgaact
     5161 tgtggccgtt tacgtcgccg tccagctcga ccaggatggg caccaccccg gtgaacagct
     5221 cctcgccctt gctcaccatg gtggcgaccg gtagcgctag cttcgaaaga tctggatccc
     5281 agaagcaaat gtaagcaata gatggctctg ccctgacttt tatgcccagc ccgagctccg
     5341 ccggcgcaca caattaagga gtaactctgg ccaaaactga gtttgcaaag ctggtctaga
     5401 gccaatgaga agcaacatca acatgaatct cctttggttt ttctctgtag caccagaaat
     5461 aatgattgca agtttgaagc ctttactgtg aaagtcaaag gtaggtgaga gccctgttct
     5521 ataacagaca ctcaccaggc actaaatccc atctttcagt atctatcttg tttgatgacc
     5581 atcaaactgc ttcctgtgga acttcacgta tctaccttca tagaccccag aggacattaa
     5641 agagagacca acagccaact catgacagcc tcatttaact gaatgatgaa gcctctccag
     5701 taagcaggaa ggtaattcaa agaagatctt atggaaatgt ttcacaaaca tcacaggtta
     5761 cagtgcagct tccaggactc tacttccaaa tccattggat cagagttttt ctgagtagat
     5821 ctcaggaaga atcattttga tcaacttatt ataaactgaa gacacacata tacataaact
     5881 tttaagcata gtagagatag tagtagatgc cttcatgatt agtatatgca ggtcacattc
     5941 aaggggtaca gaatttatat ggctttaggt tcttatgtat tagctttatc ataagacagt
     6001 tgtttataaa attgcttggt ttagaaaatg gtattcctac ttaaaatatt ttgttattgt
     6061 ccatatatgg atgtgtgtgc atatgagtga aagcacatgt gcaccataca tgtgcaggtg
     6121 acatctgcag gtaagcagac aactc
//
