LOCUS       sequence-461861-        4777 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0127m 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 4777)
  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 4777)
  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: AiP12826 - pAAV-AiE0127m-minBG-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..4777
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      789..798
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(795..797,798..800,801..1511)
                     /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(840..861)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1101..1120)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1448..1469
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1604..1624)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1811..1922
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2040..2180
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2040..2169
                     /label=AAV2 ITR
     rep_origin      2255..2710
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2342..2361)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2552..2573
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2727..2746)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2846..2868
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2906..2924)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2992..3096
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3097..3165,3166..3957)
                     /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(3315..3334)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4128..4716
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4617..4636
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtatcctg aggcttgagc tagacacctg gagattcaag
      181 cttacatgca acaacaaagg aaagtactta aaattctagc aagttaacaa atagccattt
      241 aatcccttgc tagcataatt tcctattcac atctaagatg ccaatttcaa tcatataaat
      301 ataagaaata aagagatttc tattctcaag gcatcataac tccttctcta gacctttggt
      361 gatgtaaatg aagtctctga gactgtcata ctgaggaatg atgttcacac tccattgtgt
      421 gaaatctggg ctctacattt ctgatattta tgatgcaaat gctagcaggc aaagtgcaag
      481 tgaaatcaat atgattttat ttagtattca ggcaacaggc tccatctgta caatattctc
      541 caagatgata tttaacagaa ctatagaaac taatgactgc tcagatataa atgacacttg
      601 attttaactg aaaagcaatc ttacaacacc aaagaagtgc cagtcatttt cattctaaag
      661 aatgccctgt gtatttaaag caccaaaagc ccggagagct cgggctgggc ataaaagtca
      721 gggcagagcc atctattgct tacatttgct tctgggatcc agatctttcg aagctagcgc
      781 taccggtcgc caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
      841 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg
      901 gcgatgccac ctacggcaag ctgaccctga agctgatctg caccaccggc aagctgcccg
      961 tgccctggcc caccctcgtg accaccctgg gctacggcgt gcagtgcttc gcccgctacc
     1021 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
     1081 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
     1141 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
     1201 acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat atcaccgccg
     1261 acaagcagaa gaacggcatc aaggccaact tcaagatccg ccacaacatc gaggacggcg
     1321 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
     1381 tgcccgacaa ccactacctg agctaccagt ccaagctgag caaagacccc aacgagaagc
     1441 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
     1501 agctgtacaa gtaagtcgac ggcgcgccgc ggccgcgaat tcgatatcat aatcaacctc
     1561 tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc
     1621 tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca
     1681 ttttctcctc cttgtataaa tcctggttag ttcttgccac ggcggaactc atcgccgcct
     1741 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggctcgag
     1801 agatcttcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     1861 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     1921 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     1981 ggggaggatt gggaagacaa tagcaggcat gagatctcac gtgcggaccg agcggccgca
     2041 ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc
     2101 cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg
     2161 agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     2221 tatttcacac cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc
     2281 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cttagcgccc
     2341 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     2401 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     2461 aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     2521 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     2581 ctcaactcta tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggtctat
     2641 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     2701 tttacaattt tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     2761 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     2821 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     2881 tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     2941 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     3001 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     3061 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     3121 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     3181 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     3241 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     3301 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     3361 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     3421 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     3481 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     3541 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     3601 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     3661 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     3721 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     3781 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     3841 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     3901 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     3961 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     4021 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     4081 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     4141 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     4201 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     4261 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     4321 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     4381 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     4441 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     4501 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     4561 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     4621 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     4681 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     4741 cggttcctgg ccttttgctg gccttttgct cacatgt
//
