LOCUS       sequence-473481-        4584 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0017m 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 4584)
  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 4584)
  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: AiP11274 - scAAV-AiE0017m-minBG-SYFP2-WPRE3-BG...
FEATURES             Location/Qualifiers
     source          1..4584
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      574..583
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(580..582,583..585,586..1296)
                     /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(625..646)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(886..905)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1233..1254
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1389..1409)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1589..1700
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(1939..1956)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1939..1955)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1948..1970)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2164..2183)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2283..2305
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2343..2361)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2429..2533
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2534..2602,2603..3394)
                     /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(2752..2771)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3565..4153
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4054..4073
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    4441..4462
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4477..4482,4483..4500,4501..4507)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4515..4531
                     /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     4520..4542
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4539..4555
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4539..4555
                     /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 gttgtgtgtg
      181 gtcttaggga cagggggtgg ctggcaggca ggcagtggcg ggaggcacag ctgtgcagga
      241 ggacggactg ctggcatcag caagcgcccc tcctcccccc cacactaaac aatcacggca
      301 caaatatgca aatggtataa ttactgttat ttgttttgct gatataagtg tttgaaatgc
      361 aaatgtcaac tctgggcgct gttggttttt cccacccctc tctcccggac atttgcaagt
      421 cgatcagtga gccctcatcc tgggaacact aggggtgcct ccttctgaca gacttggagc
      481 gagctcgggc tgggcataaa agtcagggca gagccatcta ttgcttacat ttgcttctgg
      541 gatccagatc tttcgaagct agcgctaccg gtcgccacca tggtgagcaa gggcgaggag
      601 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
      661 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagctg
      721 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgggctac
      781 ggcgtgcagt gcttcgcccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
      841 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
      901 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
      961 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
     1021 agccacaacg tctatatcac cgccgacaag cagaagaacg gcatcaaggc caacttcaag
     1081 atccgccaca acatcgagga cggcggcgtg cagctcgccg accactacca gcagaacacc
     1141 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcta ccagtccaag
     1201 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     1261 gccgggatca ctctcggcat ggacgagctg tacaagtaag tcgacggcgc gccgcggccg
     1321 cgaattcgat atcataatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     1381 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     1441 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttagttctt
     1501 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     1561 ggcactgaca attccgtggc tcgagcgact gtgccttcta gttgccagcc atctgttgtt
     1621 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     1681 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     1741 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatga ctagtccact
     1801 ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg
     1861 ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gagagggaca gatccgggcc
     1921 cgcatgcgtc gacaattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg
     1981 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
     2041 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgcct
     2101 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatat ggtgcactct
     2161 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc
     2221 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     2281 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa
     2341 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     2401 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     2461 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     2521 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     2581 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     2641 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     2701 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     2761 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     2821 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     2881 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     2941 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     3001 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     3061 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     3121 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     3181 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     3241 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     3301 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     3361 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     3421 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     3481 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     3541 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     3601 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     3661 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     3721 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     3781 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     3841 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     3901 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     3961 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     4021 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     4081 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     4141 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     4201 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     4261 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     4321 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     4381 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     4441 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
     4501 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga
     4561 ttacgccaag ctctcgagat ctag
//
