LOCUS       sequence-438953-        4842 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2339m 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 4842)
  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 4842)
  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: AiP1657 - pAAV-AiE2339m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4842
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(173..277)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     345..363
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(401..423)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     523..542
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(559..1014)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(696..717)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     908..927
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   1089..1229
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1100..1229
                     /label=AAV2 ITR
     polyA_signal    1347..1458
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     1645..1665
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(1758..2468,2469..2471,2472..2474))
                     /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(1800..1821)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2149..2168
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2408..2429
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2471..2480
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   3204..3333
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(3395..3983)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3475..3494)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4154..4842,1..103,104..172))
                     /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     4777..4796
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
       61 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
      121 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
      181 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
      241 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
      301 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
      361 ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
      421 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
      481 cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg
      541 tactgagagt gcaccataaa attgtaaacg ttaatatttt gttaaaattc gcgttaaatt
      601 tttgttaaat cagctcattt tttaaccaat agaccgaaat cggcaaaatc ccttataaat
      661 caaaagaata gcccgagata gagttgagtg ttgttccagt ttggaacaag agtccactat
      721 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
      781 tacgtgaacc atcacccaaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
      841 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
      901 gaaaggaagg gaagaaagcg aaaggagcgg gcgctaaggc gctggcaagt gtagcggtca
      961 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtactatg
     1021 gttgctttga cgtatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat
     1081 caggcgcccc tgcaggcagc tgcgcgctcg ctcgctcact gaggccgccc gggcaaagcc
     1141 cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg
     1201 agtggccaac tccatcacta ggggttcctg cggccgctcg gtccgcacgt gagatctcat
     1261 gcctgctatt gtcttcccaa tcctccccct tgctgtcctg ccccacccca ccccccagaa
     1321 tagaatgaca cctactcaga caatgcgatg caatttcctc attttattag gaaaggacag
     1381 tgggagtggc accttccagg gtcaaggaag gcacggggga ggggcaaaca acagatggct
     1441 ggcaactaga aggcacagtc gaagatctct cgagccacgg aattgtcagt gcccaacagc
     1501 cgagcccctg tccagcagcg ggcaaggcag gcggcgatga gttccgccgt ggcaagaact
     1561 aaccaggatt tatacaagga ggagaaaatg aaagccatac gggaagcaat agcatgatac
     1621 aaaggcatta aagcagcgta tccacatagc gtaaaaggag caacatagtt aagaatacca
     1681 gtcaatcttt cacaaatttt gtaatccaga ggttgattat gatatcgaat tcgcggccgc
     1741 ggcgcgccgt cgacttactt gtacagctcg tccatgccga gagtgatccc ggcggcggtc
     1801 acgaactcca gcaggaccat gtgatcgcgc ttctcgttgg ggtctttgct cagcttggac
     1861 tggtagctca ggtagtggtt gtcgggcagc agcacggggc cgtcgccgat gggggtgttc
     1921 tgctggtagt ggtcggcgag ctgcacgccg ccgtcctcga tgttgtggcg gatcttgaag
     1981 ttggccttga tgccgttctt ctgcttgtcg gcggtgatat agacgttgtg gctgttgtag
     2041 ttgtactcca gcttgtgccc caggatgttg ccgtcctcct tgaagtcgat gcccttcagc
     2101 tcgatgcggt tcaccagggt gtcgccctcg aacttcacct cggcgcgggt cttgtagttg
     2161 ccgtcgtcct tgaagaagat ggtgcgctcc tggacgtagc cttcgggcat ggcggacttg
     2221 aagaagtcgt gctgcttcat gtggtcgggg tagcgggcga agcactgcac gccgtagccc
     2281 agggtggtca cgagggtggg ccagggcacg ggcagcttgc cggtggtgca gatcagcttc
     2341 agggtcagct tgccgtaggt ggcatcgccc tcgccctcgc cggacacgct gaacttgtgg
     2401 ccgtttacgt cgccgtccag ctcgaccagg atgggcacca ccccggtgaa cagctcctcg
     2461 cccttgctca ccatggtggc gaccggtagc gctagcttcg aaagatctgg atcccagaag
     2521 caaatgtaag caatagatgg ctctgccctg acttttatgc ccagcccgag ctccggagac
     2581 aagatatatg attttttgat ttgtcttttt gctcaaattg aacacattct agagtagcca
     2641 agattgtgtg gtactgtaag atacaatgaa tctgggcact gagcttttgt tcctccacat
     2701 gagagaagaa agatgatttc ccattcagtg ttttgggtaa agactccatc tacatttcca
     2761 actgtgtgct gactcatttc cccagccaca tatgccattt tgacatttag gtttctgggt
     2821 tgaaatgtta tagactggtt cactgtgtgc tcttctctgt gcctcagacg tagacagagt
     2881 gagcatgatg tctaacagta cagaatgagc aattaaaaat tttggcatga gaatttaagg
     2941 aatgtaccac aaaatgtcac gtcctttttc ctatcatgtt tcttttctta agcaatgact
     3001 caatcccagt tcagcactct cttctaaagt cagttacaag ggctttcagc ttcattcata
     3061 tgacgtacat aactcagatc ataggttagg aaaagggctt atacattcat tctattgtaa
     3121 ctaaactttt tatttattct gtcaacacat tattgttttt aaatgtgttt gtcgattatt
     3181 actcttacaa cgcgtgcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct
     3241 gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggcggcct
     3301 cagtgagcga gcgagcgcgc agctgcctgc aggacatgtg agcaaaaggc cagcaaaagg
     3361 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     3421 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     3481 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     3541 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     3601 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     3661 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     3721 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     3781 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     3841 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     3901 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     3961 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     4021 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     4081 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     4141 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     4201 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     4261 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     4321 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     4381 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     4441 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     4501 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     4561 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     4621 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     4681 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     4741 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     4801 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aa
//
