LOCUS       sequence-449639-        4866 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0140h is an enhancer sequence, designed to drive AAV-mediated 
            transgene expression in cortical Pvalb+ neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4866)
  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 4866)
  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: AiP12547 -
            pAAV-hsA2-AiE0140h-minRho-mTFP1-WPR...
FEATURES             Location/Qualifiers
     source          1..4866
                     /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      912..921
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             918..1628
                     /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"
     primer_bind     complement(1693..1713)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1900..2011
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2129..2269
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2129..2258
                     /label=AAV2 ITR
     rep_origin      2344..2799
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2431..2450)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2641..2662
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2816..2835)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2935..2957
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2995..3013)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3081..3185
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3186..3254,3255..4046)
                     /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(3404..3423)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4217..4805
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4706..4725
                     /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 gcggccgcaa cgcgtttaga acaatggctg gcccatagta aatgccgtgt
      181 tagtgtgtta gttgctgttc ttccacgtca gaagaggcac agacaaatta ccaccaggtg
      241 gcgctcagag tctgcggagg catcacaaca gccctgaatt tgaatcctgc tctgccactg
      301 cctagttgag accttttact acctgactag ctgtttgtgt attttaggtg tttgtttatc
      361 tatacctctg agaaagccat tttttttaac atgaagaagt ataaaatatt gtgtgtgaag
      421 attaagccct ttcttgctct atgtatttga gcctagaaaa agacacactc ccaagaaatt
      481 agaataaagg gaaaaatgac tttgagatgt actttcttaa aataactgaa gtaatgaccg
      541 catgagccag ctgatgggtt tttaaatgga tacgtttcta tcagcctgct gcttgatgcc
      601 agagccaaat atatatggag tgtattatat catatcctgt aggcaggaga ctgttggggc
      661 attgggggac ttagagaggt gaagtggcag gcttggcaca ggaattaaca gcatggctac
      721 cgtcaaccag aattcgatat cataatcaac cataggtacc gagctcggga ttcagccggg
      781 agcttaggga ggggaggtca cttcataagg gcctgggggg ggagttggag ccacgagtcg
      841 tccagccgga gccccgtgtg gctgagctcc ggcctcagaa gcatccccgg gttggatcct
      901 tcgaagctag cgccaccatg gtgagcaagg gcgaggagac cacaatgggc gtaatcaagc
      961 ccgacatgaa gatcaagctg aagatggagg gcaacgtgaa tggccacgcc ttcgtgatcg
     1021 agggcgaggg cgagggcaag ccctacgacg gcaccaacac catcaacctg gaggtgaagg
     1081 agggagcccc cctgcccttc tcctacgaca ttctgaccac cgcgttcgcc tacggcaaca
     1141 gggccttcac caagtacccc gacgacatcc ccaactactt caagcagtcc ttccccgagg
     1201 gctactcttg ggagcgcacc atgaccttcg aggacaaggg catcgtgaag gtgaagtccg
     1261 acatctccat ggaggaggac tccttcatct acgagataca cctcaagggc gagaacttcc
     1321 cccccaacgg ccccgtgatg cagaagaaga ccaccggctg ggacgcctcc accgagagga
     1381 tgtacgtgcg cgacggcgtg ctgaagggcg acgtcaagca caagctgctg ctggagggcg
     1441 gcggccacca ccgcgttgac ttcaagacca tctacagggc caagaaggcg gtgaagctgc
     1501 ccgactatca ctttgtggac caccgcatcg agatcctgaa ccacgacaag gactacaaca
     1561 aggtgaccgt ttacgagagc gccgtggccc gcaactccac cgacggcatg gacgagctgt
     1621 acaagtaagt cgacatcata atcaacctct ggattacaaa atttgtgaaa gattgactgg
     1681 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     1741 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttagt
     1801 tcttgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     1861 gttgggcact gacaattccg tggctcgaga gatcttcgac tgtgccttct agttgccagc
     1921 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     1981 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     2041 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg
     2101 agatctcacg tgcggaccga gcggccgcag gaacccctag tgatggagtt ggccactccc
     2161 tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc
     2221 tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg
     2281 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat
     2341 agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     2401 ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct tcctttctcg
     2461 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     2521 ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg
     2581 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     2641 gtggactctt gttccaaact ggaacaacac tcaactctat ctcgggctat tcttttgatt
     2701 tataagggat tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat
     2761 ttaacgggaa ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa
     2821 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
     2881 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     2941 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg
     3001 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     3061 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     3121 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     3181 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     3241 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     3301 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     3361 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     3421 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     3481 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     3541 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     3601 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     3661 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     3721 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     3781 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     3841 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     3901 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     3961 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     4021 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     4081 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     4141 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     4201 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     4261 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     4321 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     4381 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     4441 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     4501 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     4561 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     4621 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     4681 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     4741 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     4801 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     4861 acatgt
//
