LOCUS       sequence-466260-        4582 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2141m 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 4582)
  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 4582)
  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: AiP1430 - pAAV-AiE2141m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4582
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      162..750
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     651..670
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   812..941
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      1405..1414
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1411..1413,1414..1416,1417..2127)
                     /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(1456..1477)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1717..1736)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2064..2085
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2220..2240)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2427..2538
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2656..2796
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2656..2785
                     /label=AAV2 ITR
     rep_origin      2871..3326
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2958..2977)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3168..3189
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3343..3362)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3462..3484
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3522..3540)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3608..3712
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3713..3781,3782..4573)
                     /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(3931..3950)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
       61 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
      121 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
      181 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
      241 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
      301 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
      361 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
      421 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
      481 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
      541 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
      601 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
      661 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
      721 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
      781 ctggcctttt gctggccttt tgctcacatg tcctgcaggc agctgcgcgc tcgctcgctc
      841 actgaggccg cccgggcgtc gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc
      901 gcgcagagag ggagtggcca actccatcac taggggttcc tgcggccgca cgcgttgtaa
      961 gagtaataat cgccggcgag ggagacacac acagacacac acacacagag agagagagag
     1021 agagagagag acagacagac agacagacag actgactgac tgactgactg agggagcagc
     1081 tggagctcaa gccccagctc atgtggaagt atccaagctg ctgttcctcg tttggttttg
     1141 gcatgaagca ctgtagtgta gtagtggaaa gaatggaagc gatgcactct tctgttacat
     1201 ctaaccacta cattaaaaat ggatatcctg atacctgacc cgaggcttga cacaaaacag
     1261 actcacttta caagcactgc aggaaatctc agaaaagtgg agacgccggc ggagctcggg
     1321 ctgggcataa aagtcagggc agagccatct attgcttaca tttgcttctg ggatccagat
     1381 ctttcgaagc tagcgctacc ggtcgccacc atggtgagca agggcgagga gctgttcacc
     1441 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
     1501 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagct gatctgcacc
     1561 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgggcta cggcgtgcag
     1621 tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     1681 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
     1741 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     1801 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac
     1861 gtctatatca ccgccgacaa gcagaagaac ggcatcaagg ccaacttcaa gatccgccac
     1921 aacatcgagg acggcggcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     1981 gacggccccg tgctgctgcc cgacaaccac tacctgagct accagtccaa gctgagcaaa
     2041 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     2101 actctcggca tggacgagct gtacaagtaa gtcgacggcg cgccgcggcc gcgaattcga
     2161 tatcataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     2221 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     2281 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttagttct tgccacggcg
     2341 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     2401 aattccgtgg ctcgagagat cttcgactgt gccttctagt tgccagccat ctgttgtttg
     2461 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     2521 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     2581 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgaga tctcacgtgc
     2641 ggaccgagcg gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc
     2701 gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg
     2761 cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc
     2821 ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg
     2881 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     2941 cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     3001 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     3061 gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg
     3121 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     3181 ccaaactgga acaacactca actctatctc gggctattct tttgatttat aagggatttt
     3241 gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     3301 taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc
     3361 cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     3421 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     3481 gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt
     3541 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     3601 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     3661 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     3721 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     3781 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     3841 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     3901 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     3961 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     4021 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     4081 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     4141 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     4201 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     4261 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     4321 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     4381 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     4441 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     4501 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     4561 taagcattgg taactgtcag ac
//
