LOCUS       sequence-460665-        5251 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0078h_3xC2 is an enhancer sequence, designed to drive 
            AAV-mediated transgene expression in specific brain cell types.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5251)
  AUTHORS   Ben-Simon Y, Hooper M, Narayan S, Daigle T, Dwivedi D, Way SW, Oster
            A, Stafford DA, Mich JK, Taormina MJ, Martinez RA, Opitz-Araya X, 
            Roth JR, Allen S, Ayala A, Bakken TE, Barcelli T, Barta S, Bendrick 
            J, Bertagnolli D, Bowlus J, Boyer G, Brouner K, Casian B, Casper T, 
            Chakka AB, Chakrabarty R, Chance RK, Chavan S, Departee M, Donadio 
            N, Dotson N, Egdorf T, Gabitto M, Garcia J, Gary A, Gasperini M, 
            Goldy J, Gore BB, Graybuck L, Greisman N, Haeseleer F, Halterman C, 
            Helback O, Hockemeyer D, Huang C, Huff S, Hunker A, Johansen N, 
            Juneau Z, Kalmbach B, Khem S, Kussick E, Kutsal R, Larsen R, Lee C, 
            Lee AY, Leibly M, Lenz GH, Liang E, Lusk N, Malone J, Mollenkopf T, 
            Morin E, Newman D, Ng L, Ngo K, Omstead V, Oyama A, Pham T, Pom CA, 
            Potekhina L, Ransford S, Rette D, Rimorin C, Rocha D, Ruiz A, 
            Sanchez REA, Sedeno-Cortes A, Sevigny JP, Shapovalova N, Shulga L, 
            Sigler AR, Siverts LA, Somasundaram S, Stewart K, Szelenyi E, Tieu 
            M, Trader C, van Velthoven CTJ, Walker M, 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 J, Tasic B
  TITLE     A suite of enhancer AAVs and transgenic mouse lines for genetic 
            access to cortical cell types (preprint).
  JOURNAL   bioRxiv [Preprint]. 2024 Sep 26:2024.06.10.597244. doi: 
            10.1101/2024.06.10.597244.
  PUBMED    38915722
REFERENCE   2  (bases 1 to 5251)
  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: "bioRxiv 
            [Preprint]. 2024 Sep 26:2024.06.10.597244. doi: 
            10.1101/2024.06.10.597244."
COMMENT     Sequence Label: AiP15150 -
            pAAV-AiE0078h_3xC2-minBG-mTFP1-10aa...
FEATURES             Location/Qualifiers
     source          1..5251
                     /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      864..873
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             870..1577
                     /codon_start=1
                     /product="monomeric teal (cyan) fluorescent protein (Ai et 
                     al., 2006)"
                     /label=mTFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEETTMGVIKPDMKIKLKMEGNVNGHAFVIEGEGEGKPYDG
                     TNTINLEVKEGAPLPFSYDILTTAFAYGNRAFTKYPDDIPNYFKQSFPEGYSWERTMTF
                     EDKGIVKVKSDISMEEDSFIYEIHLKGENFPPNGPVMQKKTTGWDASTERMYVRDGVLK
                     GDVKHKLLLEGGGHHRVDFKTIYRAKKAVKLPDYHFVDHRIEILNHDKDYNKVTVYESA
                     VARNSTDGMDELYK"
     CDS             1608..1985
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(2078..2098)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2285..2396
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2514..2654
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2514..2643
                     /label=AAV2 ITR
     rep_origin      2729..3184
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2816..2835)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3026..3047
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3201..3220)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3320..3342
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3380..3398)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3466..3570
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3571..3639,3640..4431)
                     /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(3789..3808)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4602..5190
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5091..5110
                     /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 gcgtattagc aatttggcag gtacccgagg gccataccta
      181 atctgcataa aatgaagcag attgcaaccg ccctcatctt ttttattttt aaactggttt
      241 ttgaagcaga gcataaaatc tcagagggag agacagaaga tgctagtgca tacattttcc
      301 ttcatgcctt tattttcatt ctttttgcac aaaccatctt cctgaatggc tgtttaatta
      361 gcaatttggc aggtacccga gggccatacc taatctgcat aaaatgaagc agattgcaac
      421 cgccctcatc ttttttattt ttaaactggt ttttgaagca gagcataaaa tctcagaggg
      481 agagacagaa gatgctagtg catacatttt ccttcatgcc tttattttca ttctttttgc
      541 acaaaccatc ttcctgaatg gctgtttaat tagcaatttg gcaggtaccc gagggccata
      601 cctaatctgc ataaaatgaa gcagattgca accgccctca tcttttttat ttttaaactg
      661 gtttttgaag cagagcataa aatctcagag ggagagacag aagatgctag tgcatacatt
      721 ttccttcatg cctttatttt cattcttttt gcacaaacca tcttcctgaa tggctgttta
      781 gagctcgggc tgggcataaa agtcagggca gagccatcta ttgcttacat ttgcttctgg
      841 gatccagatc tttcgaagct agcgccacca tggtgagcaa gggcgaggag accacaatgg
      901 gcgtaatcaa gcccgacatg aagatcaagc tgaagatgga gggcaacgtg aatggccacg
      961 ccttcgtgat cgagggcgag ggcgagggca agccctacga cggcaccaac accatcaacc
     1021 tggaggtgaa ggagggagcc cccctgccct tctcctacga cattctgacc accgcgttcg
     1081 cctacggcaa cagggccttc accaagtacc ccgacgacat ccccaactac ttcaagcagt
     1141 ccttccccga gggctactct tgggagcgca ccatgacctt cgaggacaag ggcatcgtga
     1201 aggtgaagtc cgacatctcc atggaggagg actccttcat ctacgagata cacctcaagg
     1261 gcgagaactt cccccccaac ggccccgtga tgcagaagaa gaccaccggc tgggacgcct
     1321 ccaccgagag gatgtacgtg cgcgacggcg tgctgaaggg cgacgtcaag cacaagctgc
     1381 tgctggaggg cggcggccac caccgcgttg acttcaagac catctacagg gccaagaagg
     1441 cggtgaagct gcccgactat cactttgtgg accaccgcat cgagatcctg aaccacgaca
     1501 aggactacaa caaggtgacc gtttacgaga gcgccgtggc ccgcaactcc accgacggca
     1561 tggacgagct gtacaagtcc ggactcagat ctggaggctc cggaggccca gagccagcga
     1621 agtctgctcc cgccccgaaa aagggctcca agaaggcggt gactaaggcg cagaagaaag
     1681 gcggcaagaa gcgcaagcgc agccgcaagg agagctattc catctatgtg tataaggttc
     1741 tgaagcaggt ccaccctgac accggcattt cgtccaaggc catgggcatc atgaactcgt
     1801 ttgtgaacga cattttcgag cgcatcgcag gtgaggcttc ccgcctggcg cattacaaca
     1861 agcgctcgac catcacctcc agggagatcc agacggccgt gcgcctgctg ctgcctgggg
     1921 agttggccaa gcacgccgtg tccgagggta ctaaggccat caccaagtac accagcgcta
     1981 agtaatgagt cgacggcgcg ccgcggccgc gaattcgata tcataatcaa cctctggatt
     2041 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     2101 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
     2161 cctccttgta taaatcctgg ttagttcttg ccacggcgga actcatcgcc gcctgccttg
     2221 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggct cgagagatct
     2281 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     2341 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     2401 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     2461 gattgggaag acaatagcag gcatgagatc tcacgtgcgg accgagcggc cgcaggaacc
     2521 cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg
     2581 accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg
     2641 cagctgcctg caggggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     2701 acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg
     2761 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccttagc gcccgctcct
     2821 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     2881 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     2941 gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     3001 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     3061 tctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggt ctattggtta
     3121 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca
     3181 attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga
     3241 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     3301 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     3361 aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     3421 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     3481 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     3541 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     3601 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     3661 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     3721 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     3781 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     3841 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     3901 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     3961 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     4021 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     4081 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     4141 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     4201 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     4261 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     4321 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     4381 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     4441 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     4501 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     4561 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     4621 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     4681 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     4741 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     4801 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     4861 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     4921 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     4981 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     5041 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     5101 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     5161 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     5221 ctggcctttt gctggccttt tgctcacatg t
//
