LOCUS       sequence-470610-        5404 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0943m 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 5404)
  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 5404)
  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: AiP13296 -
            pAAV-AiE0943m-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5404
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(71..90)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(750..1541,1542..1610))
                     /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     1373..1392
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1611..1715)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1783..1801
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1839..1861)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1961..1980
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(1997..2452)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2134..2155)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2346..2365
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2527..2667
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2538..2667
                     /label=AAV2 ITR
     polyA_signal    2785..2896
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3083..3103
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3190..3567)
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     CDS             complement(3598..3663)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             complement(3676..3732)
                     /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(3742..4452,4453..4455,4456..4458))
                     /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(3784..3805)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4133..4152
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     regulatory      4455..4464
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   5204..5333
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(join(5395..5404,1..579))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
       61 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
      121 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
      181 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
      241 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
      301 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
      361 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
      421 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
      481 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
      541 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
      601 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
      661 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
      721 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
      781 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
      841 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
      901 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
      961 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     1021 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     1081 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     1141 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     1201 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     1261 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     1321 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     1381 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     1441 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     1501 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     1561 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     1621 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     1681 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     1741 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     1801 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     1861 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     1921 tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta
     1981 ctgagagtgc accataaaat tgtaaacgtt aatattttgt taaaattcgc gttaaatttt
     2041 tgttaaatca gctcattttt taaccaatag accgaaatcg gcaaaatccc ttataaatca
     2101 aaagaatagc ccgagataga gttgagtgtt gttccagttt ggaacaagag tccactatta
     2161 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     2221 cgtgaaccat cacccaaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     2281 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     2341 aaggaaggga agaaagcgaa aggagcgggc gctaaggcgc tggcaagtgt agcggtcacg
     2401 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtactatggt
     2461 tgctttgacg tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca
     2521 ggcgcccctg caggcagctg cgcgctcgct cgctcactga ggccgcccgg gcaaagcccg
     2581 ggcgtcgggc gacctttggt cgcccggcct cagtgagcga gcgagcgcgc agagagggag
     2641 tggccaactc catcactagg ggttcctgcg gccgctcggt ccgcacgtga gatctcatgc
     2701 ctgctattgt cttcccaatc ctcccccttg ctgtcctgcc ccaccccacc ccccagaata
     2761 gaatgacacc tactcagaca atgcgatgca atttcctcat tttattagga aaggacagtg
     2821 ggagtggcac cttccagggt caaggaaggc acgggggagg ggcaaacaac agatggctgg
     2881 caactagaag gcacagtcga agatctctcg agccacggaa ttgtcagtgc ccaacagccg
     2941 agcccctgtc cagcagcggg caaggcaggc ggcgatgagt tccgccgtgg caagaactaa
     3001 ccaggattta tacaaggagg agaaaatgaa agccatacgg gaagcaatag catgatacaa
     3061 aggcattaaa gcagcgtatc cacatagcgt aaaaggagca acatagttaa gaataccagt
     3121 caatctttca caaattttgt aatccagagg ttgattatga tatcgaattc gcggccgcgg
     3181 cgcgcctcat tacttagcgc tggtgtactt ggtgatggcc ttagtaccct cggacacggc
     3241 gtgcttggcc aactccccag gcagcagcag gcgcacggcc gtctggatct cccgggaggt
     3301 gatggtcgag cgcttgttgt aatgcgccag gcgggaagcc tctcctgcga tgcgctcgaa
     3361 aatgtcgttc acaaacgaat tcatgatgcc catggccttg gacgaaatgc cggtgtcagg
     3421 gtggacctgc ttcagaacct tgtacacata gatggaatag ctctccttgc ggctgcgctt
     3481 gcgcttcttg ccgcctttct tctgcgcctt agtcaccgcc ttcttggagc cctttttcgg
     3541 ggcgggagca gacttcgctg gctctgggcc tccggagcct ccagatctga gtccggactt
     3601 gtcatcgtca tccttgtaat cgatgtcatg atctttataa tctccgtcat ggtctttgta
     3661 gtctccacta gcaccaggcc ctggattctc ctctacatcc cctgcctgtt tcaacagtga
     3721 gaagttggta gcaccagagc ccttgtacag ctcgtccatg ccgagagtga tcccggcggc
     3781 ggtcacgaac tccagcagga ccatgtgatc gcgcttctcg ttggggtctt tgctcagctt
     3841 ggactggtag ctcaggtagt ggttgtcggg cagcagcacg gggccgtcgc cgatgggggt
     3901 gttctgctgg tagtggtcgg cgagctgcac gccgccgtcc tcgatgttgt ggcggatctt
     3961 gaagttggcc ttgatgccgt tcttctgctt gtcggcggtg atatagacgt tgtggctgtt
     4021 gtagttgtac tccagcttgt gccccaggat gttgccgtcc tccttgaagt cgatgccctt
     4081 cagctcgatg cggttcacca gggtgtcgcc ctcgaacttc acctcggcgc gggtcttgta
     4141 gttgccgtcg tctttgaaga agatggtgcg ctcctggacg tagccttcgg gcatggcgga
     4201 cttgaagaag tcgtgctgct tcatgtggtc ggggtagcgg gcgaagcact gcacgccgta
     4261 gcccagggtg gtcacgaggg tgggccaggg cacgggcagc ttgccggtgg tgcagatcag
     4321 cttcagggtc agcttgccgt aggtggcatc gccctcgccc tcgccggaca cgctgaactt
     4381 gtggccgttg acatcgccgt ccagctcgac caggatgggc accaccccgg tgaacagctc
     4441 ctcgcccttg gagaccatgg tggcgaccgg tagcgctagc ttcgaaagat ctggatccca
     4501 gaagcaaatg taagcaatag atggctctgc cctgactttt atgcccagcc cgagctcagc
     4561 tctctctgaa ctcctctgtg gttgatgaat ttcattctta aaatttgcaa gaatccaaag
     4621 gccaatgact cagtgaaaat tctgtgtggc tatgagcatc caactcctta actcctgaat
     4681 tgaaagccca cccaagagtc aagaaaactg tacaaccctc aaagacaacc ccaaaatttt
     4741 cccataccag tatatgaata ctctttcccc ttgatgaaat gtaattaaga ctggagagca
     4801 tagctctttc atttggatct gtaatgtctg taggtgttgt tctcagcact ggatttacaa
     4861 gctctgcacc caaaaaaagg tggcttaatg cttctgagca gtagtttcaa tgtctgtaag
     4921 aaagaaagat acagagtagg ttgaacataa tgaataaaat atgcagcatc ctgactcaaa
     4981 cacaggaaac attgaacaat agcagtttac ctttataatc agagaaatga tgtaaaaggg
     5041 ttattggttc atcatttaat taattctcta agtgaattat tcccacacaa cctagagtta
     5101 ggcagagccc agggtgtcaa gaatgaactc accacttgct accatataac aagagaaata
     5161 aagactgact tgctagagct tagccgctaa cgcgtgcggc cgcaggaacc cctagtgatg
     5221 gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc
     5281 gcccgacgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggacatgtg
     5341 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     5401 tagg
//
