LOCUS       sequence-461944-        4809 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2150m 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 4809)
  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 4809)
  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: AiP1379 - pAAV-AiE2150m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4809
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   651..780
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      1471..1480
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1477..1479,1480..1482,1483..2193)
                     /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(1522..1543)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1783..1802)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2130..2151
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2286..2306)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2493..2604
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2722..2862
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2722..2851
                     /label=AAV2 ITR
     rep_origin      2937..3392
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3024..3043)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3234..3255
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3409..3428)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3528..3550
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3588..3606)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3674..3778
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3779..3847,3848..4639)
                     /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(3997..4016)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt cctgcaggca
      661 gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt ggtcgcccgg
      721 cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact aggggttcct
      781 gcggccgcac gcgtcgcttt acccagccgt atctcatttt tagacattta agttacttcc
      841 aacttttcct gcttatcagt tacgtcagca tggctatttt tatacctaga tggcctgtgt
      901 tcctgactgc ttggaagtag acttactggg tcaaagggga tgccagtgtg attttcagaa
      961 aacgatgccc gtttctgttc acagctggtt tctataaagc atgggaacgg cgcaggcact
     1021 atttatgtgc cctgctgaat tctcagctcc ctgcttcatc tgaccttccg ccactactca
     1081 gaatcggctc tgttcttctc ctgggctccc gatgggagcc aagctggact tgtgtcagtg
     1141 tattaagaag tgctcgttat agctcattgc ccatgggatg tcgactgctg ctgatttctc
     1201 agggctggag atgacacatt ttctcacatt cttcaaaagt cttaaggttc agcaggagag
     1261 aaggaaggtc cgagccaagg agagctgcgg ccataccgac agggagggca ctctctcttt
     1321 ctctagctct cccccccccc ccttgaaaag gggagaaaag ctccctccct gcctcctgag
     1381 ctcgggctgg gcataaaagt cagggcagag ccatctattg cttacatttg cttctgggat
     1441 ccagatcttt cgaagctagc gctaccggtc gccaccatgg tgagcaaggg cgaggagctg
     1501 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
     1561 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagctgatc
     1621 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gggctacggc
     1681 gtgcagtgct tcgcccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
     1741 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
     1801 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
     1861 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc
     1921 cacaacgtct atatcaccgc cgacaagcag aagaacggca tcaaggccaa cttcaagatc
     1981 cgccacaaca tcgaggacgg cggcgtgcag ctcgccgacc actaccagca gaacaccccc
     2041 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagctacca gtccaagctg
     2101 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     2161 gggatcactc tcggcatgga cgagctgtac aagtaagtcg acggcgcgcc gcggccgcga
     2221 attcgatatc ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     2281 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     2341 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt agttcttgcc
     2401 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     2461 actgacaatt ccgtggctcg agagatcttc gactgtgcct tctagttgcc agccatctgt
     2521 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     2581 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     2641 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgagatctc
     2701 acgtgcggac cgagcggccg caggaacccc tagtgatgga gttggccact ccctctctgc
     2761 gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc
     2821 gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt
     2881 ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac catagtacgc
     2941 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     3001 acttgccagc gccttagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     3061 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     3121 tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc
     3181 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     3241 cttgttccaa actggaacaa cactcaactc tatctcgggc tattcttttg atttataagg
     3301 gattttgccg atttcggtct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     3361 gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta caatctgctc
     3421 tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg
     3481 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     3541 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg
     3601 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     3661 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     3721 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     3781 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     3841 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     3901 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     3961 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     4021 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     4081 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     4141 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     4201 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     4261 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     4321 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     4381 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     4441 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     4501 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     4561 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     4621 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     4681 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     4741 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     4801 aggatcttc
//
