LOCUS       sequence-473483-        4874 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0031m 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 4874)
  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 4874)
  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: AiP11288 - scAAV-AiE0031m-minBG-SYFP2-WPRE3-BG...
FEATURES             Location/Qualifiers
     source          1..4874
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      864..873
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(870..872,873..875,876..1586)
                     /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(915..936)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1176..1195)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1523..1544
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1679..1699)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1879..1990
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2229..2246)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2229..2245)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2238..2260)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2454..2473)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2573..2595
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2633..2651)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2719..2823
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2824..2892,2893..3684)
                     /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(3042..3061)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3855..4443
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4344..4363
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    4731..4752
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4767..4772,4773..4790,4791..4797)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4805..4821
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     4810..4832
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4829..4845
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4829..4845
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aaagcttccc ggggggatct gggccactcc ctctctgcgc gctcgctcgc tcactgaggc
       61 cgcccgggca aagcccgggc ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc
      121 aactccatca ctaggggttc ctggaggggt ggagtcgtga cctaggacgc gttagggctg
      181 acagtggaaa tgagcctagc tgtcaggtgg caggagtgtg gcttccctcc ttcatagcaa
      241 acaataatac gtggctcaag gtgtgtctct atggatttgt gaagtataga aaagtggctg
      301 ccagagtagt cccatcagtg gtagccaagg atgcaccttt ggaaattgtg gagctagggc
      361 agcacatgtg aagaactccc aagaagatac gatttattct gtgctgtttg tttaagcaca
      421 tgaatttcct aatggagcca tcagaccttg cttattggct aatagtttaa ttaggcccat
      481 ctggaggatg ctgtggtgaa aactgcccca gctctgatag tggcagggat gtaatttgtc
      541 attctgtggc caatgactgg cacactccca gccctaatgc catgcaggag tagtgctgac
      601 tgtctcatgg ggagacagat agcacctgag acattttgtg gttcaccttg gtgcagaagg
      661 aattacaggt gatactcaaa gtccttgagc taacctcaac aaggtaagac agactttcta
      721 aaagtcacac ggagactgct gtcagccatc ccaggagaaa gacttttcca gagctcgggc
      781 tgggcataaa agtcagggca gagccatcta ttgcttacat ttgcttctgg gatccagatc
      841 tttcgaagct agcgctaccg gtcgccacca tggtgagcaa gggcgaggag ctgttcaccg
      901 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
      961 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagctg atctgcacca
     1021 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgggctac ggcgtgcagt
     1081 gcttcgcccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
     1141 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
     1201 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
     1261 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg
     1321 tctatatcac cgccgacaag cagaagaacg gcatcaaggc caacttcaag atccgccaca
     1381 acatcgagga cggcggcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
     1441 acggccccgt gctgctgccc gacaaccact acctgagcta ccagtccaag ctgagcaaag
     1501 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
     1561 ctctcggcat ggacgagctg tacaagtaag tcgacggcgc gccgcggccg cgaattcgat
     1621 atcataatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     1681 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     1741 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttagttctt gccacggcgg
     1801 aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca
     1861 attccgtggc tcgagcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     1921 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     1981 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     2041 acagcaaggg ggaggattgg gaagacaata gcaggcatga ctagtccact ccctctctgc
     2101 gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc
     2161 gggcggcctc agtgagcgag cgagcgcgca gagagggaca gatccgggcc cgcatgcgtc
     2221 gacaattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
     2281 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
     2341 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgcct gatgcggtat
     2401 tttctcctta cgcatctgtg cggtatttca caccgcatat ggtgcactct cagtacaatc
     2461 tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc
     2521 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     2581 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg
     2641 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     2701 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     2761 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     2821 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     2881 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     2941 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     3001 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     3061 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     3121 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     3181 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     3241 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     3301 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     3361 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     3421 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     3481 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     3541 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     3601 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     3661 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     3721 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     3781 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     3841 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     3901 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     3961 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     4021 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     4081 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     4141 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     4201 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     4261 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     4321 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     4381 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     4441 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     4501 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     4561 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     4621 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     4681 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
     4741 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     4801 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag
     4861 ctctcgagat ctag
//
