LOCUS       sequence-470727-        5228 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0551h 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 5228)
  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 5228)
  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: AiP13654 -
            pAAV-AiE0551h-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5228
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    236..347
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     534..554
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(641..1018)
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     CDS             complement(1049..1114)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             complement(1127..1183)
                     /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(1193..1903,1904..1906,1907..1909))
                     /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(1235..1256)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1584..1603
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     regulatory      1906..1915
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   2479..2608
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(2670..3258)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2750..2769)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3429..4220,4221..4289))
                     /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     4052..4071
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4290..4394)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4462..4480
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4518..4540)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4640..4659
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(4676..5131)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4813..4834)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5025..5044
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   join(5206..5228,1..118)
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   join(5217..5228,1..118)
                     /label=AAV2 ITR
ORIGIN
        1 tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg cgacctttgg tcgcccggcc
       61 tcagtgagcg agcgagcgcg cagagaggga gtggccaact ccatcactag gggttcctgc
      121 ggccgctcgg tccgcacgtg agatctcatg cctgctattg tcttcccaat cctccccctt
      181 gctgtcctgc cccaccccac cccccagaat agaatgacac ctactcagac aatgcgatgc
      241 aatttcctca ttttattagg aaaggacagt gggagtggca ccttccaggg tcaaggaagg
      301 cacgggggag gggcaaacaa cagatggctg gcaactagaa ggcacagtcg aagatctctc
      361 gagccacgga attgtcagtg cccaacagcc gagcccctgt ccagcagcgg gcaaggcagg
      421 cggcgatgag ttccgccgtg gcaagaacta accaggattt atacaaggag gagaaaatga
      481 aagccatacg ggaagcaata gcatgataca aaggcattaa agcagcgtat ccacatagcg
      541 taaaaggagc aacatagtta agaataccag tcaatctttc acaaattttg taatccagag
      601 gttgattatg atatcgaatt cgcggccgcg gcgcgcctca ttacttagcg ctggtgtact
      661 tggtgatggc cttagtaccc tcggacacgg cgtgcttggc caactcccca ggcagcagca
      721 ggcgcacggc cgtctggatc tcccgggagg tgatggtcga gcgcttgttg taatgcgcca
      781 ggcgggaagc ctctcctgcg atgcgctcga aaatgtcgtt cacaaacgaa ttcatgatgc
      841 ccatggcctt ggacgaaatg ccggtgtcag ggtggacctg cttcagaacc ttgtacacat
      901 agatggaata gctctccttg cggctgcgct tgcgcttctt gccgcctttc ttctgcgcct
      961 tagtcaccgc cttcttggag ccctttttcg gggcgggagc agacttcgct ggctctgggc
     1021 ctccggagcc tccagatctg agtccggact tgtcatcgtc atccttgtaa tcgatgtcat
     1081 gatctttata atctccgtca tggtctttgt agtctccact agcaccaggc cctggattct
     1141 cctctacatc ccctgcctgt ttcaacagtg agaagttggt agcaccagag cccttgtaca
     1201 gctcgtccat gccgagagtg atcccggcgg cggtcacgaa ctccagcagg accatgtgat
     1261 cgcgcttctc gttggggtct ttgctcagct tggactggta gctcaggtag tggttgtcgg
     1321 gcagcagcac ggggccgtcg ccgatggggg tgttctgctg gtagtggtcg gcgagctgca
     1381 cgccgccgtc ctcgatgttg tggcggatct tgaagttggc cttgatgccg ttcttctgct
     1441 tgtcggcggt gatatagacg ttgtggctgt tgtagttgta ctccagcttg tgccccagga
     1501 tgttgccgtc ctccttgaag tcgatgccct tcagctcgat gcggttcacc agggtgtcgc
     1561 cctcgaactt cacctcggcg cgggtcttgt agttgccgtc gtctttgaag aagatggtgc
     1621 gctcctggac gtagccttcg ggcatggcgg acttgaagaa gtcgtgctgc ttcatgtggt
     1681 cggggtagcg ggcgaagcac tgcacgccgt agcccagggt ggtcacgagg gtgggccagg
     1741 gcacgggcag cttgccggtg gtgcagatca gcttcagggt cagcttgccg taggtggcat
     1801 cgccctcgcc ctcgccggac acgctgaact tgtggccgtt gacatcgccg tccagctcga
     1861 ccaggatggg caccaccccg gtgaacagct cctcgccctt ggagaccatg gtggcgaccg
     1921 gtagcgctag cttcgaaaga tctggatccc agaagcaaat gtaagcaata gatggctctg
     1981 ccctgacttt tatgcccagc ccgagctcga ttcgtccggt ggacaccact catgacgggt
     2041 attacagagg catagtggta acacctgatg ggaatttcca tcgtggatat cacctccaag
     2101 tgttaggtaa ggagcatcac atactgaggt atttcacatc accagatttg tgactgaaac
     2161 aaatgtctct ctgaattcca tatgctttat gtgaaaactg aagcattttt ttccctcttt
     2221 cctctctgca gttacacccg aagtgaacct atttcaaagc aggaatataa ctgcagtatg
     2281 caaggcagtt acagggaagc cagctgccca gatctcctgg atcccagagg gatctattct
     2341 tgccactaag caagaatact ggggcaatgg cacagtgacg gttaagagta catgcccctg
     2401 ggagggccac aagtctactg tgacctgcca tgtctcccat ttgactggca acaagagtct
     2461 gtccacgcgt gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg
     2521 ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc ggcctcagtg
     2581 agcgagcgag cgcgcagctg cctgcaggac atgtgagcaa aaggccagca aaaggccagg
     2641 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     2701 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     2761 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     2821 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     2881 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     2941 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     3001 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     3061 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     3121 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     3181 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     3241 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     3301 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     3361 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     3421 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     3481 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     3541 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     3601 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     3661 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     3721 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     3781 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     3841 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     3901 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     3961 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     4021 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     4081 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     4141 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     4201 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     4261 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     4321 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     4381 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt
     4441 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc
     4501 ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg
     4561 taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt
     4621 cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccataaaatt
     4681 gtaaacgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     4741 aaccaataga ccgaaatcgg caaaatccct tataaatcaa aagaatagcc cgagatagag
     4801 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     4861 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc acccaaatca
     4921 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     4981 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     5041 ggagcgggcg ctaaggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     5101 gccgcgctta atgcgccgct acagggcgcg tactatggtt gctttgacgt atgcggtgtg
     5161 aaataccgca cagatgcgta aggagaaaat accgcatcag gcgcccctgc aggcagctgc
     5221 gcgctcgc
//
