LOCUS       sequence-466255-        5441 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2170m 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 5441)
  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 5441)
  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: AiP1399 - pAAV-AiE2170m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..5441
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   455..584
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(646..1234)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(726..745)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1405..2196,2197..2265))
                     /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     2028..2047
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2266..2370)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2438..2456
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2494..2516)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2616..2635
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2652..3107)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2789..2810)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3001..3020
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   3182..3322
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3193..3322
                     /label=AAV2 ITR
     polyA_signal    3440..3551
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3738..3758
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(3851..4561,4562..4564,4565..4567))
                     /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(3893..3914)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4242..4261
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4501..4522
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4564..4573
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 tcttagaata atgcagacac tgctggcaca gaataaacca agaatgcatg agtttagtgg
       61 gcttaataaa tctgactggt aaaaaagata tgaggttaaa aaaaaaaaaa actaagagga
      121 aatccatatc aatgtataca gatacttgaa gaaaaactag aattacaaaa ctttcctttc
      181 aagaggttgg agatggtgca atggttaaga gcatgatatg ctcttacaga ggaccttagg
      241 tttccttacc gagcagatta tgtcctcctg gaaccaatcc caaggattac ccaccccttc
      301 acccccattt tgaacaccct ggcaccagcc caccaatgtg cacataccca ctgttgtcaa
      361 atcccttttt taaagggaaa ctacgcagta tcaactcatc ttcctaaact tccaatgaca
      421 aactgaagca caactctacc acgcgtgcgg ccgcaggaac ccctagtgat ggagttggcc
      481 actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc
      541 ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg caggacatgt gagcaaaagg
      601 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
      661 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
      721 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
      781 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
      841 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
      901 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
      961 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     1021 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     1081 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     1141 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     1201 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     1261 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     1321 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     1381 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     1441 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     1501 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     1561 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     1621 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     1681 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     1741 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     1801 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     1861 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     1921 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     1981 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     2041 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     2101 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     2161 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     2221 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     2281 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     2341 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     2401 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
     2461 tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac
     2521 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
     2581 gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag
     2641 agtgcaccat aaaattgtaa acgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     2701 aatcagctca ttttttaacc aatagaccga aatcggcaaa atcccttata aatcaaaaga
     2761 atagcccgag atagagttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     2821 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     2881 accatcaccc aaatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     2941 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     3001 agggaagaaa gcgaaaggag cgggcgctaa ggcgctggca agtgtagcgg tcacgctgcg
     3061 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtact atggttgctt
     3121 tgacgtatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc
     3181 ccctgcaggc agctgcgcgc tcgctcgctc actgaggccg cccgggcaaa gcccgggcgt
     3241 cgggcgacct ttggtcgccc ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc
     3301 aactccatca ctaggggttc ctgcggccgc tcggtccgca cgtgagatct catgcctgct
     3361 attgtcttcc caatcctccc ccttgctgtc ctgccccacc ccacccccca gaatagaatg
     3421 acacctactc agacaatgcg atgcaatttc ctcattttat taggaaagga cagtgggagt
     3481 ggcaccttcc agggtcaagg aaggcacggg ggaggggcaa acaacagatg gctggcaact
     3541 agaaggcaca gtcgaagatc tctcgagcca cggaattgtc agtgcccaac agccgagccc
     3601 ctgtccagca gcgggcaagg caggcggcga tgagttccgc cgtggcaaga actaaccagg
     3661 atttatacaa ggaggagaaa atgaaagcca tacgggaagc aatagcatga tacaaaggca
     3721 ttaaagcagc gtatccacat agcgtaaaag gagcaacata gttaagaata ccagtcaatc
     3781 tttcacaaat tttgtaatcc agaggttgat tatgatatcg aattcgcggc cgcggcgcgc
     3841 cgtcgactta cttgtacagc tcgtccatgc cgagagtgat cccggcggcg gtcacgaact
     3901 ccagcaggac catgtgatcg cgcttctcgt tggggtcttt gctcagcttg gactggtagc
     3961 tcaggtagtg gttgtcgggc agcagcacgg ggccgtcgcc gatgggggtg ttctgctggt
     4021 agtggtcggc gagctgcacg ccgccgtcct cgatgttgtg gcggatcttg aagttggcct
     4081 tgatgccgtt cttctgcttg tcggcggtga tatagacgtt gtggctgttg tagttgtact
     4141 ccagcttgtg ccccaggatg ttgccgtcct ccttgaagtc gatgcccttc agctcgatgc
     4201 ggttcaccag ggtgtcgccc tcgaacttca cctcggcgcg ggtcttgtag ttgccgtcgt
     4261 ccttgaagaa gatggtgcgc tcctggacgt agccttcggg catggcggac ttgaagaagt
     4321 cgtgctgctt catgtggtcg gggtagcggg cgaagcactg cacgccgtag cccagggtgg
     4381 tcacgagggt gggccagggc acgggcagct tgccggtggt gcagatcagc ttcagggtca
     4441 gcttgccgta ggtggcatcg ccctcgccct cgccggacac gctgaacttg tggccgttta
     4501 cgtcgccgtc cagctcgacc aggatgggca ccaccccggt gaacagctcc tcgcccttgc
     4561 tcaccatggt ggcgaccggt agcgctagct tcgaaagatc tggatcccag aagcaaatgt
     4621 aagcaataga tggctctgcc ctgactttta tgcccagccc gagctcgctt acttcacaga
     4681 gcaaagagta cttgtatttt accaaattat ctagaaggat taaagactta cacatacagt
     4741 gtattagata ctaaaatgtg aatactagca aaaatgtttc catccttcag tcattgatct
     4801 cctgtatcta gagtcttcat ttttagggac aaagcctcca aatttccaat cttttaactt
     4861 ataagcaatc catattttca gtgggagctg ctaaaaaata tttgtacttc taaaacaatt
     4921 tttttggttc aattcaaagc tatatgcatt caatacatgc tagcaatgga tgcccgttta
     4981 ttaactatag ggtctaaagc acatcacctc atttcttaca gtcattgttt gaggtgggag
     5041 gaacagttca gcaaggaaac agctgcaaga acaatggaca aatctgaagc tagcgaaatg
     5101 tacagtgcaa ctaacaatta catcaagtca caaagcttca tctatgacag gtgtgctcca
     5161 aagcgactgt tctgtgcgat gacacctaca gaaagctagt ttaacaggga ggcatgcttt
     5221 attattaact gagtggggtg tgggactcct agaaacaaag tagtagttaa ctgagatgat
     5281 acgggcttat tttcttcttc tgccgacccc ctgcttttcg tcttattctt cctttccttc
     5341 actgttggag gagcatccta ataaaaacat cgaagtgaca ctgttatcac actctctgct
     5401 tttcttcatg ctcatgattc catgactaac aggacagatg g
//
