LOCUS       sequence-466232-        4948 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0058m 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 4948)
  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 4948)
  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: AiP1275 - pAAV-AiE0058m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4948
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      368..377
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(374..376,377..379,380..1090)
                     /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(419..440)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(680..699)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1027..1048
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1183..1203)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1390..1501
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1619..1759
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1619..1748
                     /label=AAV2 ITR
     rep_origin      1834..2289
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1921..1940)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2131..2152
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2306..2325)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2425..2447
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2485..2503)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2571..2675
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2676..2744,2745..3536)
                     /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(2894..2913)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3707..4295
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4196..4215
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   4357..4486
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 tcatctactt ccatcttgtg aaaagtttcc actcttcagt gatgacctca aacatatcat
       61 ttcttttttt taattctttt tttattagat attttcttta tatacatttc aaatgctatc
      121 ctgaaagttc cctatatcct ccctctgccc tgcttcccta cccacccact cccacttctt
      181 ggccctggca tttccctgta ctggggcata taaagtttgc aataccaagg ggcctctctt
      241 cacagtgatg gccaactagg ccatcttctg ctacgagctc gggctgggca taaaagtcag
      301 ggcagagcca tctattgctt acatttgctt ctgggatcca gatctttcga agctagcgct
      361 accggtcgcc accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
      421 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg
      481 cgatgccacc tacggcaagc tgaccctgaa gctgatctgc accaccggca agctgcccgt
      541 gccctggccc accctcgtga ccaccctggg ctacggcgtg cagtgcttcg cccgctaccc
      601 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
      661 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
      721 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
      781 catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata tcaccgccga
      841 caagcagaag aacggcatca aggccaactt caagatccgc cacaacatcg aggacggcgg
      901 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
      961 gcccgacaac cactacctga gctaccagtc caagctgagc aaagacccca acgagaagcg
     1021 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
     1081 gctgtacaag taagtcgacg gcgcgccgcg gccgcgaatt cgatatcata atcaacctct
     1141 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     1201 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     1261 tttctcctcc ttgtataaat cctggttagt tcttgccacg gcggaactca tcgccgcctg
     1321 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggctcgaga
     1381 gatcttcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     1441 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     1501 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     1561 gggaggattg ggaagacaat agcaggcatg agatctcacg tgcggaccga gcggccgcag
     1621 gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc
     1681 gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga
     1741 gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     1801 atttcacacc gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg
     1861 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg
     1921 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     1981 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     2041 aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     2101 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     2161 tcaactctat ctcgggctat tcttttgatt tataagggat tttgccgatt tcggtctatt
     2221 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt
     2281 ttacaatttt atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc
     2341 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     2401 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
     2461 caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca
     2521 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     2581 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     2641 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     2701 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     2761 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     2821 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     2881 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     2941 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     3001 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     3061 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     3121 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     3181 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     3241 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     3301 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     3361 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     3421 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     3481 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     3541 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     3601 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     3661 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     3721 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     3781 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     3841 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     3901 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     3961 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     4021 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     4081 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     4141 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     4201 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     4261 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     4321 ggttcctggc cttttgctgg ccttttgctc acatgtcctg caggcagctg cgcgctcgct
     4381 cgctcactga ggccgcccgg gcgtcgggcg acctttggtc gcccggcctc agtgagcgag
     4441 cgagcgcgca gagagggagt ggccaactcc atcactaggg gttcctgcgg ccgcacgcgt
     4501 ggtacatatt ataagtttga gtctgcaaga tgtggcaaac ccttcctttt ctctttatct
     4561 tgacagtgga aaacatctaa ggagtcttta aaataaccac atcggactga gcagtggagg
     4621 gcaaagagaa tgtctggaag aacctttgtt tttttcgttg ggacatcaaa ggatgttata
     4681 ctgaagagat ctcaaagacc aggcagtcca gcctactcgt ttcccagaca agagaagtga
     4741 agccaagaaa cagatgggct gcctcctcta gtcatatggg gcttaaaaat atatatctct
     4801 atttttcatg agttttcatg aaagtgacca gtgttatctc atttcctgtc ttctcaaatt
     4861 caggatccta cccaatctaa ctattattta tagtgtgaat aagccctcta ctttcctacc
     4921 tctacaatgc tcattctcca ttttccca
//
