LOCUS       sequence-470766-        5283 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE1126m 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 5283)
  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 5283)
  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: AiP14289 -
            pAAV-AiE1126m-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5283
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(43..834,835..903))
                     /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     666..685
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(904..1008)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1076..1094
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1132..1154)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1254..1273
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(1290..1745)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1427..1448)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1639..1658
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     misc_feature    1814..1841
                     /gene="telL"
                     /label=telL
                     /note="telomeric sequence from coliphage N15"
     misc_feature    1842..1869
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     repeat_region   1876..2016
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1887..2016
                     /label=AAV2 ITR
     polyA_signal    2134..2245
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2432..2452
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2539..2916)
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     CDS             complement(2947..3012)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             complement(3025..3081)
                     /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             complement(join(3091..3801,3802..3804,3805..3807))
                     /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(3133..3154)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3482..3501
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     regulatory      3804..3813
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   4319..4448
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     misc_feature    4455..4482
                     /gene="telL"
                     /label=telL
                     /note="telomeric sequence from coliphage N15"
     misc_feature    4483..4510
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     rep_origin      complement(4567..5155)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4647..4666)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
       61 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
      121 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
      181 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
      241 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
      301 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
      361 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
      421 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
      481 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
      541 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
      601 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
      661 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
      721 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
      781 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
      841 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
      901 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
      961 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     1021 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     1081 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     1141 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     1201 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc
     1261 agagcagatt gtactgagag tgcaccataa aattgtaaac gttaatattt tgttaaaatt
     1321 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa tagaccgaaa tcggcaaaat
     1381 cccttataaa tcaaaagaat agcccgagat agagttgagt gttgttccag tttggaacaa
     1441 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     1501 cgatggccca ctacgtgaac catcacccaa atcaagtttt ttggggtcga ggtgccgtaa
     1561 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     1621 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaagg cgctggcaag
     1681 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     1741 cgcgtactat ggttgctttg acgtatgcgg tgtgaaatac cgcacagatg cgtaaggaga
     1801 aaataccgca tcatatcagc acacaatagt ccattatacg cgcgtataat gggcaattgt
     1861 gtgctgatag gcgcccctgc aggcagctgc gcgctcgctc gctcactgag gccgcccggg
     1921 caaagcccgg gcgtcgggcg acctttggtc gcccggcctc agtgagcgag cgagcgcgca
     1981 gagagggagt ggccaactcc atcactaggg gttcctgcgg ccgctcggtc cgcacgtgag
     2041 atctcatgcc tgctattgtc ttcccaatcc tcccccttgc tgtcctgccc caccccaccc
     2101 cccagaatag aatgacacct actcagacaa tgcgatgcaa tttcctcatt ttattaggaa
     2161 aggacagtgg gagtggcacc ttccagggtc aaggaaggca cgggggaggg gcaaacaaca
     2221 gatggctggc aactagaagg cacagtcgaa gatctctcga gccacggaat tgtcagtgcc
     2281 caacagccga gcccctgtcc agcagcgggc aaggcaggcg gcgatgagtt ccgccgtggc
     2341 aagaactaac caggatttat acaaggagga gaaaatgaaa gccatacggg aagcaatagc
     2401 atgatacaaa ggcattaaag cagcgtatcc acatagcgta aaaggagcaa catagttaag
     2461 aataccagtc aatctttcac aaattttgta atccagaggt tgattatgat atcgaattcg
     2521 cggccgcggc gcgcctcatt acttagcgct ggtgtacttg gtgatggcct tagtaccctc
     2581 ggacacggcg tgcttggcca actccccagg cagcagcagg cgcacggccg tctggatctc
     2641 cctggaggtg atggtcgagc gcttgttgta atgcgccagg cgggaagcct cacctgcgat
     2701 gcgctcgaaa atgtcgttca caaacgagtt catgatgccc atggccttgg acgaaatgcc
     2761 ggtgtcaggg tggacctgct tcagaacctt atacacatag atggaatagc tctccttgcg
     2821 gctgcgcttg cgcttcttgc cgcctttctt ctgcgcctta gtcaccgcct tcttggagcc
     2881 ctttttcggg gcgggagcag acttcgctgg ctctgggcct ccggagcctc cagatctgag
     2941 tccggacttg tcatcgtcat ccttgtaatc gatgtcatga tctttataat ctccgtcatg
     3001 gtctttgtag tctccactag caccaggccc tggattctcc tctacatccc ctgcctgttt
     3061 caacagtgag aagttggtag caccagagcc cttgtacagc tcgtccatgc cgagagtgat
     3121 cccggcggcg gtcacgaact ccagcaggac catgtgatcg cgcttctcgt tggggtcttt
     3181 gctcagcttg gactggtagc tcaggtagtg gttgtcgggc agcagcacgg ggccgtcgcc
     3241 gatgggggtg ttctgctggt agtggtcggc gagctgcacg ccgccgtcct cgatgttgtg
     3301 gcggatcttg aagttggcct tgatgccgtt cttctgcttg tcggcggtga tatagacgtt
     3361 gtggctgttg tagttgtact ccagcttgtg ccccaggatg ttgccgtcct ccttgaagtc
     3421 gatgcccttc agctcgatgc ggttcaccag ggtgtcgccc tcgaacttca cctcggcgcg
     3481 ggtcttgtag ttgccgtcgt ctttgaagaa gatggtgcgc tcctggacgt agccttcggg
     3541 catggcggac ttgaagaagt cgtgctgctt catgtggtcg gggtagcggg cgaagcactg
     3601 cacgccgtag cccagggtgg tcacgagggt gggccagggc acgggcagct tgccggtggt
     3661 gcagatcagc ttcagggtca gcttgccgta ggtggcatcg ccctcgccct cgccggacac
     3721 gctgaacttg tggccgttga catcgccgtc cagctcgacc aggatgggca ccaccccggt
     3781 gaacagctcc tcgcccttgg agaccatggt ggcgaccggt agcgctagct tcgaaagatc
     3841 tggatcccag aagcaaatgt aagcaataga tggctctgcc ctgactttta tgcccagccc
     3901 gagctctggt cttagggggt gcgcagggac cacagcacac cggtaaccca gaaatgaggc
     3961 gggtagtgtg aatgaatttc taggtatcga gtgaattttt ctaggcatct aagcttagaa
     4021 aagccaagga cgtgaagcct tagaaacgac aaggaggaga gaaaaaacga acgagtagca
     4081 acagccgccg aaatagctca gttgggagag cgttagactg aagatctaaa ggtccctggt
     4141 tcgatcccgg gtttcggcag ctcgtacttt ttctgcttta ctcacagttt atgttttcga
     4201 tttctttcta cataaaattc tggagcttcg taccctgtcg cagaaggctc tggaaattta
     4261 ctattctgtt tcctttaaga gtgaatgaac gtattcctta aagcacgcgt gcggccgcag
     4321 gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc
     4381 gggcgaccaa aggtcgcccg acgcccgggc ggcctcagtg agcgagcgag cgcgcagctg
     4441 cctgcaggac atgttatcag cacacaatag tccattatac gcgcgtataa tgggcaattg
     4501 tgtgctgata tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     4561 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     4621 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     4681 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     4741 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     4801 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     4861 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     4921 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     4981 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     5041 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     5101 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     5161 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     5221 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     5281 ttt
//
