LOCUS       sequence-466248-        4759 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2143m 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 4759)
  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 4759)
  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: AiP1372 - pAAV-AiE2143m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4759
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     595..614
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(833..937)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1005..1023
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1061..1083)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1183..1202
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(1219..1674)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1356..1377)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1568..1587
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   1749..1889
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1760..1889
                     /label=AAV2 ITR
     polyA_signal    2007..2118
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2305..2325
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(2418..3128,3129..3131,3132..3134))
                     /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(2460..2481)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2809..2828
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3068..3089
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3131..3140
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   3781..3910
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(3972..4560)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4052..4071)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4731..4759,1..763,764..832))
                     /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"
ORIGIN
        1 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
       61 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
      121 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
      181 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
      241 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
      301 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
      361 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
      421 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
      481 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
      541 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
      601 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
      661 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
      721 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
      781 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
      841 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
      901 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
      961 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
     1021 ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
     1081 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
     1141 cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg
     1201 tactgagagt gcaccataaa attgtaaacg ttaatatttt gttaaaattc gcgttaaatt
     1261 tttgttaaat cagctcattt tttaaccaat agaccgaaat cggcaaaatc ccttataaat
     1321 caaaagaata gcccgagata gagttgagtg ttgttccagt ttggaacaag agtccactat
     1381 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     1441 tacgtgaacc atcacccaaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     1501 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     1561 gaaaggaagg gaagaaagcg aaaggagcgg gcgctaaggc gctggcaagt gtagcggtca
     1621 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtactatg
     1681 gttgctttga cgtatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat
     1741 caggcgcccc tgcaggcagc tgcgcgctcg ctcgctcact gaggccgccc gggcaaagcc
     1801 cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg
     1861 agtggccaac tccatcacta ggggttcctg cggccgctcg gtccgcacgt gagatctcat
     1921 gcctgctatt gtcttcccaa tcctccccct tgctgtcctg ccccacccca ccccccagaa
     1981 tagaatgaca cctactcaga caatgcgatg caatttcctc attttattag gaaaggacag
     2041 tgggagtggc accttccagg gtcaaggaag gcacggggga ggggcaaaca acagatggct
     2101 ggcaactaga aggcacagtc gaagatctct cgagccacgg aattgtcagt gcccaacagc
     2161 cgagcccctg tccagcagcg ggcaaggcag gcggcgatga gttccgccgt ggcaagaact
     2221 aaccaggatt tatacaagga ggagaaaatg aaagccatac gggaagcaat agcatgatac
     2281 aaaggcatta aagcagcgta tccacatagc gtaaaaggag caacatagtt aagaatacca
     2341 gtcaatcttt cacaaatttt gtaatccaga ggttgattat gatatcgaat tcgcggccgc
     2401 ggcgcgccgt cgacttactt gtacagctcg tccatgccga gagtgatccc ggcggcggtc
     2461 acgaactcca gcaggaccat gtgatcgcgc ttctcgttgg ggtctttgct cagcttggac
     2521 tggtagctca ggtagtggtt gtcgggcagc agcacggggc cgtcgccgat gggggtgttc
     2581 tgctggtagt ggtcggcgag ctgcacgccg ccgtcctcga tgttgtggcg gatcttgaag
     2641 ttggccttga tgccgttctt ctgcttgtcg gcggtgatat agacgttgtg gctgttgtag
     2701 ttgtactcca gcttgtgccc caggatgttg ccgtcctcct tgaagtcgat gcccttcagc
     2761 tcgatgcggt tcaccagggt gtcgccctcg aacttcacct cggcgcgggt cttgtagttg
     2821 ccgtcgtcct tgaagaagat ggtgcgctcc tggacgtagc cttcgggcat ggcggacttg
     2881 aagaagtcgt gctgcttcat gtggtcgggg tagcgggcga agcactgcac gccgtagccc
     2941 agggtggtca cgagggtggg ccagggcacg ggcagcttgc cggtggtgca gatcagcttc
     3001 agggtcagct tgccgtaggt ggcatcgccc tcgccctcgc cggacacgct gaacttgtgg
     3061 ccgtttacgt cgccgtccag ctcgaccagg atgggcacca ccccggtgaa cagctcctcg
     3121 cccttgctca ccatggtggc gaccggtagc gctagcttcg aaagatctgg atcccagaag
     3181 caaatgtaag caatagatgg ctctgccctg acttttatgc ccagcccgag ctcggaaatg
     3241 gcttctcttc attgtccttg tcacttatcc atttagtcca tgaaacggtg acctcgtggg
     3301 tgtgggaagc acaggatgac aagggaaggg tttgtcccca gagagtccaa agctgacccc
     3361 ggggaatggc cagaagagga gcacagagga aagtaaacaa ttgtgagcat cacggagaaa
     3421 tccttggacc aaatgatgat ctgaggacga tgacgtcttg acccgagggt gggaggcaga
     3481 acagatggag gaagtaatcc aggaggcctg ctataggagg ggctaggtcc acagcccttc
     3541 tgtttcacag ggaccagaaa gaatccaccg gggagacgag ggtcagaata gtggccaagc
     3601 tgaagagaga gatccaagga tgctggcagt caattcccaa atccccagtc agctcataac
     3661 gacggtagga atgtgcttcc agaagcgcag agaggaattc tggtgcctcc ccacccctcc
     3721 ttctcctctc ctctcagcat tgaccccagc accttttgaa ggatgaacgc gtgcggccgc
     3781 aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg
     3841 ccgggcgacc aaaggtcgcc cgacgcccgg gcggcctcag tgagcgagcg agcgcgcagc
     3901 tgcctgcagg acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     3961 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     4021 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     4081 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     4141 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     4201 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     4261 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     4321 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     4381 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     4441 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     4501 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     4561 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     4621 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     4681 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     4741 ttaatcagtg aggcaccta
//
