LOCUS       sequence-462405-        5366 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE1014h_3xC3 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 5366)
  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 5366)
  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: AiP14283 -
            pAAV-AiE1014h_3xC3-minBG-SYFP2-P2A-...
FEATURES             Location/Qualifiers
     source          1..5366
                     /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      832..841
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(838..840,841..843,844..1554)
                     /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(1144..1163)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1491..1512
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1564..1620
                     /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             1633..1698
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1729..2106
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(2193..2213)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2400..2511
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2629..2769
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2629..2758
                     /label=AAV2 ITR
     rep_origin      2844..3299
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2931..2950)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3141..3162
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3316..3335)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3435..3457
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3495..3513)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3581..3685
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3686..3754,3755..4546)
                     /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(3904..3923)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4717..5305
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5206..5225
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtgaattt ttaatattcc tgtaatcatc tgcaaaatat
      181 acttgtgcac attttaccct tgagtttcat gggagagatt ttaaaggagt atttgtctcg
      241 gaactacagc ttatcactgt tggataaaga atattatctg gcctgccaag cagatattta
      301 aaaataacct aatcgcacat ttgatccagc aattcccctc ctgaattttt aatattcctg
      361 taatcatctg caaaatatac ttgtgcacat tttacccttg agtttcatgg gagagatttt
      421 aaaggagtat ttgtctcgga actacagctt atcactgttg gataaagaat attatctggc
      481 ctgccaagca gatatttaaa aataacctaa tcgcacattt gatccagcaa ttcccctcct
      541 gaatttttaa tattcctgta atcatctgca aaatatactt gtgcacattt tacccttgag
      601 tttcatggga gagattttaa aggagtattt gtctcggaac tacagcttat cactgttgga
      661 taaagaatat tatctggcct gccaagcaga tatttaaaaa taacctaatc gcacatttga
      721 tccagcaatt cccctcctga gctcgggctg ggcataaaag tcagggcaga gccatctatt
      781 gcttacattt gcttctggga tccagatctt tcgaagctag cgctaccggt cgccaccatg
      841 gtctccaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
      901 gatgtcaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
      961 aagctgaccc tgaagctgat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
     1021 gtgaccaccc tgggctacgg cgtgcagtgc ttcgcccgct accccgacca catgaagcag
     1081 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
     1141 aaagacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
     1201 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
     1261 ctggagtaca actacaacag ccacaacgtc tatatcaccg ccgacaagca gaagaacggc
     1321 atcaaggcca acttcaagat ccgccacaac atcgaggacg gcggcgtgca gctcgccgac
     1381 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     1441 ctgagctacc agtccaagct gagcaaagac cccaacgaga agcgcgatca catggtcctg
     1501 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagggctct
     1561 ggtgctacca acttctcact gttgaaacag gcaggggatg tagaggagaa tccagggcct
     1621 ggtgctagtg gagactacaa agaccatgac ggagattata aagatcatga catcgattac
     1681 aaggatgacg atgacaagtc cggactcaga tctggaggct ccggaggccc agagccagcg
     1741 aagtctgctc ccgccccgaa aaagggctcc aagaaggcgg tgactaaggc gcagaagaaa
     1801 ggcggcaaga agcgcaagcg cagccgcaag gagagctatt ccatctatgt gtataaggtt
     1861 ctgaagcagg tccaccctga caccggcatt tcgtccaagg ccatgggcat catgaactcg
     1921 tttgtgaacg acattttcga gcgcatcgca ggtgaggctt cccgcctggc gcattacaac
     1981 aagcgctcga ccatcacctc cagggagatc cagacggccg tgcgcctgct gctgcctggg
     2041 gagttggcca agcacgccgt gtccgagggt actaaggcca tcaccaagta caccagcgct
     2101 aagtaatgag gcgcgccgcg gccgcgaatt cgatatcata atcaacctct ggattacaaa
     2161 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     2221 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     2281 ttgtataaat cctggttagt tcttgccacg gcggaactca tcgccgcctg ccttgcccgc
     2341 tgctggacag gggctcggct gttgggcact gacaattccg tggctcgaga gatcttcgac
     2401 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     2461 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     2521 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     2581 ggaagacaat agcaggcatg agatctcacg tgcggaccga gcggccgcag gaacccctag
     2641 tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa
     2701 aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct
     2761 gcctgcaggg gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     2821 gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     2881 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc
     2941 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     3001 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt
     3061 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     3121 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaactctat
     3181 ctcgggctat tcttttgatt tataagggat tttgccgatt tcggtctatt ggttaaaaaa
     3241 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt
     3301 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     3361 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     3421 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     3481 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     3541 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     3601 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     3661 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     3721 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     3781 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     3841 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     3901 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     3961 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     4021 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     4081 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     4141 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     4201 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     4261 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     4321 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     4381 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     4441 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     4501 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     4561 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     4621 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     4681 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     4741 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     4801 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     4861 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     4921 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     4981 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     5041 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     5101 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     5161 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     5221 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     5281 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     5341 cttttgctgg ccttttgctc acatgt
//
