LOCUS       sequence-432539-        5875 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2135m 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 5875)
  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 5875)
  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: AiP1427 - pAAV-AiE2135m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..5875
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(47..635)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(127..146)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(806..1597,1598..1666))
                     /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     1429..1448
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1667..1771)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1839..1857
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1895..1917)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2017..2036
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2053..2508)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2190..2211)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2402..2421
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2583..2723
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2594..2723
                     /label=AAV2 ITR
     polyA_signal    2841..2952
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3139..3159
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(3252..3962,3963..3965,3966..3968))
                     /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(3294..3315)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3643..3662
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3902..3923
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3965..3974
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   5731..5860
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
       61 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
      121 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
      181 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
      241 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
      301 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
      361 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
      421 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
      481 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
      541 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
      601 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
      661 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
      721 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
      781 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
      841 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
      901 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
      961 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     1021 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     1081 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     1141 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     1201 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     1261 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     1321 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     1381 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     1441 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     1501 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     1561 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     1621 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     1681 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     1741 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     1801 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
     1861 ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga
     1921 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
     1981 cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga
     2041 gagtgcacca taaaattgta aacgttaata ttttgttaaa attcgcgtta aatttttgtt
     2101 aaatcagctc attttttaac caatagaccg aaatcggcaa aatcccttat aaatcaaaag
     2161 aatagcccga gatagagttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     2221 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     2281 aaccatcacc caaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     2341 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     2401 aagggaagaa agcgaaagga gcgggcgcta aggcgctggc aagtgtagcg gtcacgctgc
     2461 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtac tatggttgct
     2521 ttgacgtatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
     2581 cccctgcagg cagctgcgcg ctcgctcgct cactgaggcc gcccgggcaa agcccgggcg
     2641 tcgggcgacc tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc
     2701 caactccatc actaggggtt cctgcggccg ctcggtccgc acgtgagatc tcatgcctgc
     2761 tattgtcttc ccaatcctcc cccttgctgt cctgccccac cccacccccc agaatagaat
     2821 gacacctact cagacaatgc gatgcaattt cctcatttta ttaggaaagg acagtgggag
     2881 tggcaccttc cagggtcaag gaaggcacgg gggaggggca aacaacagat ggctggcaac
     2941 tagaaggcac agtcgaagat ctctcgagcc acggaattgt cagtgcccaa cagccgagcc
     3001 cctgtccagc agcgggcaag gcaggcggcg atgagttccg ccgtggcaag aactaaccag
     3061 gatttataca aggaggagaa aatgaaagcc atacgggaag caatagcatg atacaaaggc
     3121 attaaagcag cgtatccaca tagcgtaaaa ggagcaacat agttaagaat accagtcaat
     3181 ctttcacaaa ttttgtaatc cagaggttga ttatgatatc gaattcgcgg ccgcggcgcg
     3241 ccgtcgactt acttgtacag ctcgtccatg ccgagagtga tcccggcggc ggtcacgaac
     3301 tccagcagga ccatgtgatc gcgcttctcg ttggggtctt tgctcagctt ggactggtag
     3361 ctcaggtagt ggttgtcggg cagcagcacg gggccgtcgc cgatgggggt gttctgctgg
     3421 tagtggtcgg cgagctgcac gccgccgtcc tcgatgttgt ggcggatctt gaagttggcc
     3481 ttgatgccgt tcttctgctt gtcggcggtg atatagacgt tgtggctgtt gtagttgtac
     3541 tccagcttgt gccccaggat gttgccgtcc tccttgaagt cgatgccctt cagctcgatg
     3601 cggttcacca gggtgtcgcc ctcgaacttc acctcggcgc gggtcttgta gttgccgtcg
     3661 tccttgaaga agatggtgcg ctcctggacg tagccttcgg gcatggcgga cttgaagaag
     3721 tcgtgctgct tcatgtggtc ggggtagcgg gcgaagcact gcacgccgta gcccagggtg
     3781 gtcacgaggg tgggccaggg cacgggcagc ttgccggtgg tgcagatcag cttcagggtc
     3841 agcttgccgt aggtggcatc gccctcgccc tcgccggaca cgctgaactt gtggccgttt
     3901 acgtcgccgt ccagctcgac caggatgggc accaccccgg tgaacagctc ctcgcccttg
     3961 ctcaccatgg tggcgaccgg tagcgctagc ttcgaaagat ctggatccca gaagcaaatg
     4021 taagcaatag atggctctgc cctgactttt atgcccagcc cgagctccgc cggcgaaggt
     4081 ctgccgtgcc acttgatcag aagtgtgatc tgaggggagg accggacaga atgagctcct
     4141 gagatgaggc tctccgctcc aatctaacct cagtttctgg gtgactttga acacattgct
     4201 tgccctctct ctgtgccttg agagacttct gcagtgccac aggagtgaga ctaacacccc
     4261 ccccacacac acaccacccc agaagctcat gatgctgtga ctggcagagg ggaaggacaa
     4321 ataaaggtgg cttttgagtg ttatttgggc tcctcacgac ccgcctgggc accacagcca
     4381 gttctgtggc cttagtttgt gtctgggcag acttttattt aaatgccttt actaatgtta
     4441 gcttgcagtg gaatccacaa tggacaagaa agtaattggg aaatatttct tcaacaaata
     4501 aaagagccaa cggggcctgt cagttcacag cttgaaccgg gctctgggcc cagtttcctc
     4561 tctagcctct gtgagctgta aaatgagaac atgcatacat atgcacacgc acgtgtacac
     4621 gcgcatgcgc gcacgcacgc acacacacac acacaaacac acactcccct gtgatctatg
     4681 atttagagac agggtcttgc cttgaagatc cggttaactt caaactcatg attctcctgc
     4741 ctcagcatcc caagtactag aatcacagtt acatgccact gaggcctaat acgtttgcag
     4801 tctctgacgt acctgttatg gtcactaaaa tcctgcccgc tatctcatct cttgtcctgg
     4861 tgcctcctga ggcctgcctt gtccagatca ccaggtagaa gagcctgggt ctcaatgaac
     4921 tcggtgatgg gaactcgggt gggcagagaa tgaggaagga ccacagtgcc ccaaggacag
     4981 gttgctccct ctttggcccc taagtccacc ctccccccaa gtggctgggg gcctggcctg
     5041 agatcagagg tccaccagcg tcaagccgac cctgcccagc gcgtcaagag gcgccttctc
     5101 agcgcagaac aatgccggtt ctttgaagga ccgggcagca ggggatgggt gttctccaaa
     5161 cactgggcct cctttgcaaa acaggctttg attcccaggc ccagaggccc caggtgcctg
     5221 ttgggcggca gtcgggtggc tgggggcttt gtttggcagg aaagggggcc ttggagactc
     5281 aggaacaaag gcggggacag agctggatgg agttgtggtg gcctgggtag gcaatcaggc
     5341 ttacaaaggt gggggtgggg tctgtacctc tctggcagtc cctcttcggc ctgtcccact
     5401 cagggcagtc cctgttctct gtaactctgt cacggataca gcatgtgcag tcggacttcc
     5461 aggttggcct ggtagcccct gggttccaaa aaacctctaa tttggaaatg aagtcgataa
     5521 gaccggagat cagaatccaa gaagcaggag ccagagagcc tagaacggaa agtccctggc
     5581 ctgtgcccca agtcttaggg tacagtgtta aaagtgaggg gtctgatttg tgtcagatat
     5641 catgctgagt gctggagaca cctggctctc tcacttgacc cccatgttcc cgacgccggc
     5701 gattattact cttacaacgc gtgcggccgc aggaacccct agtgatggag ttggccactc
     5761 cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg
     5821 gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg acatgtgagc aaaag
//
