LOCUS       sequence-470606-        5172 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0571m 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 5172)
  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 5172)
  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: AiP12707 -
            pAAV-AiE0571m-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5172
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(150..941,942..1010))
                     /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     773..792
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1011..1115)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1183..1201
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1239..1261)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1361..1380
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(1397..1852)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1534..1555)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1746..1765
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   1927..2067
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1938..2067
                     /label=AAV2 ITR
     polyA_signal    2185..2296
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2483..2503
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2590..2967)
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     CDS             complement(2998..3063)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             complement(3076..3132)
                     /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(3142..3852,3853..3855,3856..3858))
                     /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(3184..3205)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3533..3552
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     regulatory      3855..3864
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   4372..4501
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(4563..5151)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4643..4662)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
       61 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
      121 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
      181 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
      241 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
      301 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
      361 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
      421 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
      481 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
      541 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
      601 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
      661 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
      721 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
      781 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
      841 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
      901 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
      961 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     1021 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     1081 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     1141 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     1201 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     1261 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     1321 tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta
     1381 ctgagagtgc accataaaat tgtaaacgtt aatattttgt taaaattcgc gttaaatttt
     1441 tgttaaatca gctcattttt taaccaatag accgaaatcg gcaaaatccc ttataaatca
     1501 aaagaatagc ccgagataga gttgagtgtt gttccagttt ggaacaagag tccactatta
     1561 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     1621 cgtgaaccat cacccaaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     1681 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     1741 aaggaaggga agaaagcgaa aggagcgggc gctaaggcgc tggcaagtgt agcggtcacg
     1801 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtactatggt
     1861 tgctttgacg tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca
     1921 ggcgcccctg caggcagctg cgcgctcgct cgctcactga ggccgcccgg gcaaagcccg
     1981 ggcgtcgggc gacctttggt cgcccggcct cagtgagcga gcgagcgcgc agagagggag
     2041 tggccaactc catcactagg ggttcctgcg gccgctcggt ccgcacgtga gatctcatgc
     2101 ctgctattgt cttcccaatc ctcccccttg ctgtcctgcc ccaccccacc ccccagaata
     2161 gaatgacacc tactcagaca atgcgatgca atttcctcat tttattagga aaggacagtg
     2221 ggagtggcac cttccagggt caaggaaggc acgggggagg ggcaaacaac agatggctgg
     2281 caactagaag gcacagtcga agatctctcg agccacggaa ttgtcagtgc ccaacagccg
     2341 agcccctgtc cagcagcggg caaggcaggc ggcgatgagt tccgccgtgg caagaactaa
     2401 ccaggattta tacaaggagg agaaaatgaa agccatacgg gaagcaatag catgatacaa
     2461 aggcattaaa gcagcgtatc cacatagcgt aaaaggagca acatagttaa gaataccagt
     2521 caatctttca caaattttgt aatccagagg ttgattatga tatcgaattc gcggccgcgg
     2581 cgcgcctcat tacttagcgc tggtgtactt ggtgatggcc ttagtaccct cggacacggc
     2641 gtgcttggcc aactccccag gcagcagcag gcgcacggcc gtctggatct cccgggaggt
     2701 gatggtcgag cgcttgttgt aatgcgccag gcgggaagcc tctcctgcga tgcgctcgaa
     2761 aatgtcgttc acaaacgaat tcatgatgcc catggccttg gacgaaatgc cggtgtcagg
     2821 gtggacctgc ttcagaacct tgtacacata gatggaatag ctctccttgc ggctgcgctt
     2881 gcgcttcttg ccgcctttct tctgcgcctt agtcaccgcc ttcttggagc cctttttcgg
     2941 ggcgggagca gacttcgctg gctctgggcc tccggagcct ccagatctga gtccggactt
     3001 gtcatcgtca tccttgtaat cgatgtcatg atctttataa tctccgtcat ggtctttgta
     3061 gtctccacta gcaccaggcc ctggattctc ctctacatcc cctgcctgtt tcaacagtga
     3121 gaagttggta gcaccagagc ccttgtacag ctcgtccatg ccgagagtga tcccggcggc
     3181 ggtcacgaac tccagcagga ccatgtgatc gcgcttctcg ttggggtctt tgctcagctt
     3241 ggactggtag ctcaggtagt ggttgtcggg cagcagcacg gggccgtcgc cgatgggggt
     3301 gttctgctgg tagtggtcgg cgagctgcac gccgccgtcc tcgatgttgt ggcggatctt
     3361 gaagttggcc ttgatgccgt tcttctgctt gtcggcggtg atatagacgt tgtggctgtt
     3421 gtagttgtac tccagcttgt gccccaggat gttgccgtcc tccttgaagt cgatgccctt
     3481 cagctcgatg cggttcacca gggtgtcgcc ctcgaacttc acctcggcgc gggtcttgta
     3541 gttgccgtcg tctttgaaga agatggtgcg ctcctggacg tagccttcgg gcatggcgga
     3601 cttgaagaag tcgtgctgct tcatgtggtc ggggtagcgg gcgaagcact gcacgccgta
     3661 gcccagggtg gtcacgaggg tgggccaggg cacgggcagc ttgccggtgg tgcagatcag
     3721 cttcagggtc agcttgccgt aggtggcatc gccctcgccc tcgccggaca cgctgaactt
     3781 gtggccgttg acatcgccgt ccagctcgac caggatgggc accaccccgg tgaacagctc
     3841 ctcgcccttg gagaccatgg tggcgaccgg tagcgctagc ttcgaaagat ctggatccca
     3901 gaagcaaatg taagcaatag atggctctgc cctgactttt atgcccagcc cgagctcctg
     3961 aaattactcc aaggtcaaga agataataaa ccttatatac ctggtcaaag tacaatacaa
     4021 catttgacat cagttcatgg gaatattatg ttgtcacatc actagcaaat gtgtctaaca
     4081 ttacaacatg actccatcta caaaatttac actcaagaat taaatggctg agttataaaa
     4141 agaatttcac aatattctaa gtttaggacg ttatttatac tgtattggta actgttcttg
     4201 ggcagctgtg acctatatgg ctttgggctg gcattcctat tgcattccaa aacacagaga
     4261 aatttacaga aatcctagaa aaccctctga tgatctgaat tctcacttgt tcatctttac
     4321 ttcttcattc tctattcatg atgttttctc catgaagacg cgtgcggccg caggaacccc
     4381 tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac
     4441 caaaggtcgc ccgacgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag
     4501 gacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     4561 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     4621 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     4681 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     4741 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     4801 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     4861 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     4921 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     4981 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     5041 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     5101 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     5161 ttgatctttt ct
//
