LOCUS       sequence-470780-        5542 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE1381m 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 5542)
  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 5542)
  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: AiP14977 -
            pAAV-AiE1381m-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5542
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      149..737
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     638..657
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    794..821
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     misc_feature    822..849
                     /gene="telL"
                     /label=telL
                     /note="telomeric sequence from coliphage N15"
     repeat_region   856..985
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      1750..1759
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1756..1758,1759..1761,1762..2472)
                     /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(2062..2081)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2409..2430
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2482..2538
                     /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             2551..2616
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2647..3024
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(3111..3131)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    3318..3429
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3547..3687
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3547..3676
                     /label=AAV2 ITR
     misc_feature    3694..3721
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     misc_feature    3722..3749
                     /gene="telL"
                     /label=telL
                     /note="telomeric sequence from coliphage N15"
     rep_origin      3818..4273
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3905..3924)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4115..4136
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4290..4309)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4409..4431
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4469..4487)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4555..4659
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4660..4728,4729..5520)
                     /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(4878..4897)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
       61 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
      121 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
      181 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
      241 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
      301 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
      361 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
      421 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
      481 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
      541 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
      601 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
      661 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
      721 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
      781 ggccttttgc tcatatcagc acacaattgc ccattatacg cgcgtataat ggactattgt
      841 gtgctgataa catgtcctgc aggcagctgc gcgctcgctc gctcactgag gccgcccggg
      901 cgtcgggcga cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg
      961 gccaactcca tcactagggg ttcctgcggc cgcacgcgtg tccatgtttg agggaagcca
     1021 gacaaggatc tcaggacaaa aacctagagg caggaactga agcagaagcc aaggaggagt
     1081 gcgacttact ggtttctcat ggcttgctca tcctgctttc ttatgccatc caggaccacc
     1141 caccccttgt cagatgagcc cacccacatc agacatcaat caagaaaatg ttccacagac
     1201 ttgatcacag gccaatctgg tggggacatt ttctcaattg aggtttcctc tctttcaaaa
     1261 tgactctagt ctgtgtcaag ttgacataaa aactagccag cacagatggt gactataaca
     1321 tttcctgctt cccagggaga ctgggaggat ttaatgagtc aatacacatg gttttatagt
     1381 gtggcctgcc gtctgctgac gacacacacg atagctattg tattagttac tttctagtcg
     1441 ttgcaaccaa aataccaaag gataccgtca tgtgcaggat ttgtattttg agtcacagtt
     1501 tcagaagtgt cagctcctca tggcacggag ggagtgttgg gacagaggaa atttttatca
     1561 aggcaggcag gaagcagagg tggggtgtgt gtgtgcaaat attagaatgt ggctgacttt
     1621 gatttatttt gactataccc tcagctcctg tgacccgagc tcgggctggg cataaaagtc
     1681 agggcagagc catctattgc ttacatttgc ttctgggatc cagatctttc gaagctagcg
     1741 ctaccggtcg ccaccatggt ctccaagggc gaggagctgt tcaccggggt ggtgcccatc
     1801 ctggtcgagc tggacggcga tgtcaacggc cacaagttca gcgtgtccgg cgagggcgag
     1861 ggcgatgcca cctacggcaa gctgaccctg aagctgatct gcaccaccgg caagctgccc
     1921 gtgccctggc ccaccctcgt gaccaccctg ggctacggcg tgcagtgctt cgcccgctac
     1981 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     2041 gagcgcacca tcttcttcaa agacgacggc aactacaaga cccgcgccga ggtgaagttc
     2101 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     2161 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcaccgcc
     2221 gacaagcaga agaacggcat caaggccaac ttcaagatcc gccacaacat cgaggacggc
     2281 ggcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     2341 ctgcccgaca accactacct gagctaccag tccaagctga gcaaagaccc caacgagaag
     2401 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     2461 gagctgtaca agggctctgg tgctaccaac ttctcactgt tgaaacaggc aggggatgta
     2521 gaggagaatc cagggcctgg tgctagtgga gactacaaag accatgacgg agattataaa
     2581 gatcatgaca tcgattacaa ggatgacgat gacaagtccg gactcagatc tggaggctcc
     2641 ggaggcccag agccagcgaa gtctgctccc gccccgaaaa agggctccaa gaaggcggtg
     2701 actaaggcgc agaagaaagg cggcaagaag cgcaagcgca gccgcaagga gagctattcc
     2761 atctatgtgt ataaggttct gaagcaggtc caccctgaca ccggcatttc gtccaaggcc
     2821 atgggcatca tgaactcgtt tgtgaacgac attttcgagc gcatcgcagg tgaggcttcc
     2881 cgcctggcgc attacaacaa gcgctcgacc atcacctcca gggagatcca gacggccgtg
     2941 cgcctgctgc tgcctgggga gttggccaag cacgccgtgt ccgagggtac taaggccatc
     3001 accaagtaca ccagcgctaa gtaatgaggc gcgccgcggc cgcgaattcg atatcataat
     3061 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     3121 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     3181 gctttcattt tctcctcctt gtataaatcc tggttagttc ttgccacggc ggaactcatc
     3241 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     3301 gctcgagaga tcttcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     3361 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     3421 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     3481 cagcaagggg gaggattggg aagacaatag caggcatgag atctcacgtg cggaccgagc
     3541 ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca
     3601 ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga
     3661 gcgagcgagc gcgcagctgc ctgcaggggc gcctatcagc acacaattgc ccattatacg
     3721 cgcgtataat ggactattgt gtgctgatat gatgcggtat tttctcctta cgcatctgtg
     3781 cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta
     3841 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccttagcg
     3901 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     3961 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     4021 aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     4081 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     4141 acactcaact ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggtc
     4201 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     4261 acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     4321 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     4381 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     4441 tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     4501 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     4561 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     4621 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     4681 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     4741 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     4801 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     4861 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     4921 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     4981 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     5041 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     5101 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     5161 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     5221 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     5281 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     5341 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     5401 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     5461 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     5521 ctgtcagacc aagtttactc at
//
