LOCUS       sequence-462706-        5793 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE1430m 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 5793)
  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 5793)
  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: AiP15054 -
            pAAV-AiE1430m-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5793
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      1146..1155
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1152..1154,1155..1157,1158..1868)
                     /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(1458..1477)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1805..1826
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1878..1934
                     /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             1947..2012
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2043..2420
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(2507..2527)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2714..2825
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2943..3083
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2943..3072
                     /label=AAV2 ITR
     misc_feature    3090..3117
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     misc_feature    3118..3145
                     /gene="telL"
                     /label=telL
                     /note="telomeric sequence from coliphage N15"
     rep_origin      3214..3669
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3301..3320)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3511..3532
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3686..3705)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3805..3827
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3865..3883)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3951..4055
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4056..4124,4125..4916)
                     /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(4274..4293)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5087..5675
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5576..5595
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5732..5759
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     misc_feature    5760..5787
                     /gene="telL"
                     /label=telL
                     /note="telomeric sequence from coliphage N15"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgttacaga accaagtgcc cgaccccaga gtctttcttt
      181 gtctgacctg agggctttct gttaaaatcc ttatacagca gataagtaag tcagcagtct
      241 tagaggtggg aagaggtctg tctgccagtc atctggctcc ctgcaaacac aggcatcccc
      301 acccctgaac aattttcttc ataggaaagt tgtgagggaa gtcacaggtg cagacaccgc
      361 cccggggggg gagggggggc tcagagttgt tccttcattc agtcaccaat tcagtcttcc
      421 agaatcaagg ccctattgac agagggctaa gccaccgggg cgctgccttc aaggtaagag
      481 gcatagatca gtctcctgaa tgtagaggga tgaattccct ccttcacata agtgaaagcg
      541 atcataggga acggaggcat cattagggcg ccctccttct gacagggatg cagacaggca
      601 ggtgagggtg gaggggtgtg gcccagggca cattcagagc caaactctgt tgagaacaag
      661 gcaggtgttt ctgaaaacct ctctgattgc acccgtaccc tcccatcagg cccgaatcgc
      721 acacctctct gcctctcctc agacgccggc ccacatggct tttgaaaacc tggggagatt
      781 ataccttcca gcttgctttg ctgagtctga ggaacctggc tgccttttct cttctgttaa
      841 ctgcaatcaa actggttttg agcaccagcc ccaagatgtc tttgccagtc cacatccatc
      901 ttccatgatg agctaaggtt ccctatttgt acccttactt tctgggctga gtcatgacat
      961 ctgggaatgg tgctttctct gagaacttgc ggcggcggcg gggggggggg ggagtggctg
     1021 agggggggtt acgaagacaa tgagctatgt gggagctcgg gctgggcata aaagtcaggg
     1081 cagagccatc tattgcttac atttgcttct gggatccaga tctttcgaag ctagcgctac
     1141 cggtcgccac catggtctcc aagggcgagg agctgttcac cggggtggtg cccatcctgg
     1201 tcgagctgga cggcgatgtc aacggccaca agttcagcgt gtccggcgag ggcgagggcg
     1261 atgccaccta cggcaagctg accctgaagc tgatctgcac caccggcaag ctgcccgtgc
     1321 cctggcccac cctcgtgacc accctgggct acggcgtgca gtgcttcgcc cgctaccccg
     1381 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
     1441 gcaccatctt cttcaaagac gacggcaact acaagacccg cgccgaggtg aagttcgagg
     1501 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
     1561 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc accgccgaca
     1621 agcagaagaa cggcatcaag gccaacttca agatccgcca caacatcgag gacggcggcg
     1681 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     1741 ccgacaacca ctacctgagc taccagtcca agctgagcaa agaccccaac gagaagcgcg
     1801 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
     1861 tgtacaaggg ctctggtgct accaacttct cactgttgaa acaggcaggg gatgtagagg
     1921 agaatccagg gcctggtgct agtggagact acaaagacca tgacggagat tataaagatc
     1981 atgacatcga ttacaaggat gacgatgaca agtccggact cagatctgga ggctccggag
     2041 gcccagagcc agcgaagtct gctcccgccc cgaaaaaggg ctccaagaag gcggtgacta
     2101 aggcgcagaa gaaaggcggc aagaagcgca agcgcagccg caaggagagc tattccatct
     2161 atgtgtataa ggttctgaag caggtccacc ctgacaccgg catttcgtcc aaggccatgg
     2221 gcatcatgaa ctcgtttgtg aacgacattt tcgagcgcat cgcaggtgag gcttcccgcc
     2281 tggcgcatta caacaagcgc tcgaccatca cctccaggga gatccagacg gccgtgcgcc
     2341 tgctgctgcc tggggagttg gccaagcacg ccgtgtccga gggtactaag gccatcacca
     2401 agtacaccag cgctaagtaa tgaggcgcgc cgcggccgcg aattcgatat cataatcaac
     2461 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
     2521 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
     2581 tcattttctc ctccttgtat aaatcctggt tagttcttgc cacggcggaa ctcatcgccg
     2641 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggctc
     2701 gagagatctt cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     2761 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     2821 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     2881 aagggggagg attgggaaga caatagcagg catgagatct cacgtgcgga ccgagcggcc
     2941 gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga
     3001 ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga
     3061 gcgagcgcgc agctgcctgc aggggcgcct atcagcacac aattgcccat tatacgcgcg
     3121 tataatggac tattgtgtgc tgatatgatg cggtattttc tccttacgca tctgtgcggt
     3181 atttcacacc gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg
     3241 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg
     3301 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     3361 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     3421 aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     3481 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     3541 tcaactctat ctcgggctat tcttttgatt tataagggat tttgccgatt tcggtctatt
     3601 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt
     3661 ttacaatttt atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc
     3721 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     3781 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
     3841 caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca
     3901 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     3961 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     4021 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     4081 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     4141 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     4201 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     4261 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     4321 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     4381 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     4441 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     4501 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     4561 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     4621 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     4681 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     4741 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     4801 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     4861 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     4921 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     4981 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     5041 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     5101 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     5161 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     5221 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     5281 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     5341 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     5401 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     5461 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     5521 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     5581 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     5641 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     5701 ggttcctggc cttttgctgg ccttttgctc atatcagcac acaattgccc attatacgcg
     5761 cgtataatgg actattgtgt gctgataaca tgt
//
