LOCUS       sequence-470763-        5190 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE1124m 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 5190)
  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 5190)
  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: AiP14258 -
            pAAV-AiE1124m-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5190
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      78..533
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(165..184)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     375..396
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(550..569)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     669..691
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(729..747)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        815..919
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(920..988,989..1780)
                     /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(1138..1157)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1951..2539
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2440..2459
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    2596..2623
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     misc_feature    2624..2651
                     /gene="telL"
                     /label=telL
                     /note="telomeric sequence from coliphage N15"
     repeat_region   2658..2787
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      3200..3209
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3206..3208,3209..3211,3212..3922)
                     /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(3512..3531)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3859..3880
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3932..3988
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             4001..4066
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             4097..4474
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(4561..4581)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    4768..4879
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4997..5137
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4997..5126
                     /label=AAV2 ITR
     misc_feature    5144..5171
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     misc_feature    join(5172..5190,1..9)
                     /gene="telL"
                     /label=telL
                     /note="telomeric sequence from coliphage N15"
ORIGIN
        1 gtgctgatat gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac
       61 gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
      121 tacgcgcagc gtgaccgcta cacttgccag cgccttagcg cccgctcctt tcgctttctt
      181 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
      241 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga
      301 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
      361 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaact ctatctcggg
      421 ctattctttt gatttataag ggattttgcc gatttcggtc tattggttaa aaaatgagct
      481 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg
      541 cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac
      601 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
      661 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
      721 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
      781 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
      841 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
      901 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
      961 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     1021 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     1081 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     1141 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     1201 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     1261 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     1321 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     1381 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     1441 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     1501 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     1561 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     1621 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     1681 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     1741 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     1801 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     1861 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     1921 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     1981 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     2041 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     2101 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     2161 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     2221 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     2281 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     2341 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     2401 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     2461 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     2521 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     2581 ctggcctttt gctcatatca gcacacaatt gcccattata cgcgcgtata atggactatt
     2641 gtgtgctgat aacatgtcct gcaggcagct gcgcgctcgc tcgctcactg aggccgcccg
     2701 ggcgtcgggc gacctttggt cgcccggcct cagtgagcga gcgagcgcgc agagagggag
     2761 tggccaactc catcactagg ggttcctgcg gccgcacgcg tactagccgt gctcaaaagc
     2821 ccgaggtgac tattgcggcc gataggacca cggggtcaca ggaagcagca gccggtgagg
     2881 gaccaggccc tcttcctttg tgtgggtgac tcacccgccc gctccaccgg gctgccgcgt
     2941 cctccatttt gagctccttg caacagggcc cgggagcggc catctttcca cgcacgcaac
     3001 tggtgccgga cgggatggcc tcaccctagt tagggaggca gggcaacgcg gcgccgccaa
     3061 gccagatcgt gccgggtgct ggggccacat ggcctcggca cgctaagagc tcgggctggg
     3121 cataaaagtc agggcagagc catctattgc ttacatttgc ttctgggatc cagatctttc
     3181 gaagctagcg ctaccggtcg ccaccatggt ctccaagggc gaggagctgt tcaccggggt
     3241 ggtgcccatc ctggtcgagc tggacggcga tgtcaacggc cacaagttca gcgtgtccgg
     3301 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagctgatct gcaccaccgg
     3361 caagctgccc gtgccctggc ccaccctcgt gaccaccctg ggctacggcg tgcagtgctt
     3421 cgcccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg
     3481 ctacgtccag gagcgcacca tcttcttcaa agacgacggc aactacaaga cccgcgccga
     3541 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa
     3601 ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta
     3661 tatcaccgcc gacaagcaga agaacggcat caaggccaac ttcaagatcc gccacaacat
     3721 cgaggacggc ggcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg
     3781 ccccgtgctg ctgcccgaca accactacct gagctaccag tccaagctga gcaaagaccc
     3841 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct
     3901 cggcatggac gagctgtaca agggctctgg tgctaccaac ttctcactgt tgaaacaggc
     3961 aggggatgta gaggagaatc cagggcctgg tgctagtgga gactacaaag accatgacgg
     4021 agattataaa gatcatgaca tcgattacaa ggatgacgat gacaagtccg gactcagatc
     4081 tggaggctcc ggaggcccag agccagcgaa gtctgctccc gccccgaaaa agggctccaa
     4141 gaaggcggtg actaaggcgc agaagaaagg cggcaagaag cgcaagcgca gccgcaagga
     4201 gagctattcc atctatgtgt ataaggttct gaagcaggtc caccctgaca ccggcatttc
     4261 gtccaaggcc atgggcatca tgaactcgtt tgtgaacgac attttcgagc gcatcgcagg
     4321 tgaggcttcc cgcctggcgc attacaacaa gcgctcgacc atcacctcca gggagatcca
     4381 gacggccgtg cgcctgctgc tgcctgggga gttggccaag cacgccgtgt ccgagggtac
     4441 taaggccatc accaagtaca ccagcgctaa gtaatgaggc gcgccgcggc cgcgaattcg
     4501 atatcataat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     4561 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     4621 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttagttc ttgccacggc
     4681 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     4741 caattccgtg gctcgagaga tcttcgactg tgccttctag ttgccagcca tctgttgttt
     4801 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     4861 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     4921 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgag atctcacgtg
     4981 cggaccgagc ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct
     5041 cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg
     5101 gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc gcctatcagc acacaattgc
     5161 ccattatacg cgcgtataat ggactattgt
//
