LOCUS       sequence-449790-        4827 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0354h is an enhancer sequence, designed to drive AAV-mediated 
            transgene expression in cortical Vip+ neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4827)
  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 4827)
  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: AiP14908 - pAAV-AiE0354h-minBG-mScarlet-WPRE3-...
FEATURES             Location/Qualifiers
     source          1..4827
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     regulatory      860..869
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             866..1561
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(999..1017)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1267..1286
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(1654..1674)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1861..1972
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2090..2230
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2090..2219
                     /label=AAV2 ITR
     rep_origin      2305..2760
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2392..2411)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2602..2623
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2777..2796)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2896..2918
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2956..2974)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3042..3146
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3147..3215,3216..4007)
                     /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(3365..3384)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4178..4766
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4667..4686
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtaggta cctctacatt gatggacctt
      181 tctccagaat gaggagccaa aatcatctga attaagtaga gatctcattt gtccctgact
      241 catgataatg tctgtgctac agtcctgcat taattacttt gcccaagtag aaatacacta
      301 cccatataag aatttcactt ctgaacaaaa caaaataaaa atccacgagt ggtgacctca
      361 aaatgaactg gttaaattta aggactcatt tttaaaggtc aatgtccttt tgctgcaaat
      421 acagttccta tgcagatgag aagttgaacc tgacctatgg agccgagcat aattagccag
      481 ctgcaccctc ataataaggt gcagatggtt aattgcacca gcaaataact gcttacttgt
      541 gggcacaaaa gcctgtacct aaaatcattg aaagtatact tgggactatt tatgctcatg
      601 gcaaacaaaa cacatttcca aggtttcatc tgcgaacaga tcacagccag ctccaaatcc
      661 ctgctttgcg ggagagccat tcacgctgct gaggtctggt ccaagagagg gaccactggc
      721 cacaaagcca catatcaact ggtgaaatat ttcttggggt gctgaatcgt tgttgagagc
      781 tcgggctggg cataaaagtc agggcagagc catctattgc ttacatttgc ttctgggatc
      841 cagatctttc gaagctagcg ccaccatggt gagcaagggc gaggcagtga tcaaggagtt
      901 catgcggttc aaggtgcaca tggagggctc catgaacggc cacgagttcg agatcgaggg
      961 cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg
     1021 tggccccctg cccttctcct gggacatcct gtcccctcag ttcatgtacg gctccagggc
     1081 cttcaccaag caccccgccg acatccccga ctactataag cagtccttcc ccgagggctt
     1141 caagtgggag cgcgtgatga acttcgagga cggcggcgcc gtgaccgtga cccaggacac
     1201 ctccctggag gacggcaccc tgatctacaa ggtgaagctc cgcggcacca acttccctcc
     1261 tgacggcccc gtaatgcaga agaagacaat gggctgggaa gcgtccaccg agcggttgta
     1321 ccccgaggac ggcgtgctga agggcgacat taagatggcc ctgcgcctga aggacggcgg
     1381 ccgctacctg gcggacttca agaccaccta caaggccaag aagcccgtgc agatgcccgg
     1441 cgcctacaac gtcgaccgca agttggacat cacctcccac aacgaggact acaccgtggt
     1501 ggaacagtac gaacgctccg agggccgcca ctccaccggc ggcatggacg agctgtacaa
     1561 gtaagtcgac ggcgcgccgc ggccgcgaat tcgatatcat aatcaacctc tggattacaa
     1621 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     1681 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     1741 cttgtataaa tcctggttag ttcttgccac ggcggaactc atcgccgcct gccttgcccg
     1801 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggctcgag agatcttcga
     1861 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     1921 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     1981 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     2041 gggaagacaa tagcaggcat gagatctcac gtgcggaccg agcggccgca ggaaccccta
     2101 gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca
     2161 aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc
     2221 tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     2281 cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     2341 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     2401 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     2461 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     2521 tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     2581 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     2641 tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     2701 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt
     2761 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc
     2821 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
     2881 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
     2941 gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
     3001 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     3061 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     3121 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     3181 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     3241 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     3301 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     3361 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     3421 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     3481 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     3541 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     3601 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     3661 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     3721 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     3781 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     3841 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     3901 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     3961 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     4021 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     4081 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     4141 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     4201 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     4261 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     4321 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     4381 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     4441 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     4501 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     4561 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     4621 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     4681 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     4741 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     4801 ccttttgctg gccttttgct cacatgt
//
