LOCUS       sequence-470518-        4530 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0668m 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 4530)
  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 4530)
  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: AiP12753 - pAAV-AiE0668m-minBG-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..4530
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      159..168
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(165..167,168..170,171..881)
                     /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(210..231)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(471..490)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     818..839
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(974..994)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1181..1292
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1410..1550
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1410..1539
                     /label=AAV2 ITR
     rep_origin      1625..2080
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1712..1731)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1922..1943
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2097..2116)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2216..2238
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2276..2294)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2362..2466
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2467..2535,2536..3327)
                     /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(2685..2704)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3498..4086
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3987..4006
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   4148..4277
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 ccgatctgct tcccaagcta acatatgttg gcatccaatc ctgaaaacta cagaagacat
       61 gaggtgagct cgggctgggc ataaaagtca gggcagagcc atctattgct tacatttgct
      121 tctgggatcc agatctttcg aagctagcgc taccggtcgc caccatggtg agcaagggcg
      181 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
      241 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
      301 agctgatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctgg
      361 gctacggcgt gcagtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca
      421 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
      481 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
      541 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
      601 acaacagcca caacgtctat atcaccgccg acaagcagaa gaacggcatc aaggccaact
      661 tcaagatccg ccacaacatc gaggacggcg gcgtgcagct cgccgaccac taccagcaga
      721 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agctaccagt
      781 ccaagctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
      841 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaagtcgac ggcgcgccgc
      901 ggccgcgaat tcgatatcat aatcaacctc tggattacaa aatttgtgaa agattgactg
      961 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt
     1021 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttag
     1081 ttcttgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc
     1141 tgttgggcac tgacaattcc gtggctcgag agatcttcga ctgtgccttc tagttgccag
     1201 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     1261 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     1321 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
     1381 gagatctcac gtgcggaccg agcggccgca ggaaccccta gtgatggagt tggccactcc
     1441 ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg
     1501 ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat
     1561 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgtc aaagcaacca
     1621 tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     1681 accgctacac ttgccagcgc cttagcgccc gctcctttcg ctttcttccc ttcctttctc
     1741 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     1801 tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt
     1861 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     1921 agtggactct tgttccaaac tggaacaaca ctcaactcta tctcgggcta ttcttttgat
     1981 ttataaggga ttttgccgat ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa
     2041 tttaacgcga attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca
     2101 atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg
     2161 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     2221 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc
     2281 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     2341 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     2401 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     2461 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     2521 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     2581 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     2641 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     2701 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     2761 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     2821 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     2881 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     2941 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     3001 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     3061 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     3121 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     3181 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     3241 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     3301 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     3361 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     3421 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     3481 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     3541 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     3601 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     3661 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     3721 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     3781 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     3841 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     3901 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     3961 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     4021 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     4081 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     4141 cacatgtcct gcaggcagct gcgcgctcgc tcgctcactg aggccgcccg ggcgtcgggc
     4201 gacctttggt cgcccggcct cagtgagcga gcgagcgcgc agagagggag tggccaactc
     4261 catcactagg ggttcctgcg gccgcacgcg tacacacaca cacacacaca cacacaaata
     4321 aaaacaaaaa caactcttaa aaacctaaga aagggataaa tacaacatca aagactgttg
     4381 gggttataaa acatacacat ttatagacat ttccaagatg ctactaaaat tacatacaaa
     4441 tgagtatcca aaatgtcaca gaacaaatta ttcttgtgca acataaacaa ctagagagca
     4501 tagcaaagga agcccagacc atgagctttc
//
