LOCUS       sequence-449774-        4433 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0219h 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 4433)
  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 4433)
  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: AiP15262 - pAAV-AiE0219h-minBG-mScarlet-WPRE3-...
FEATURES             Location/Qualifiers
     source          1..4433
                     /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      466..475
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             472..1167
                     /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(605..623)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     873..892
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(1260..1280)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1467..1578
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1696..1836
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1696..1825
                     /label=AAV2 ITR
     rep_origin      1911..2366
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1998..2017)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2208..2229
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2383..2402)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2502..2524
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2562..2580)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2648..2752
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2753..2821,2822..3613)
                     /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(2971..2990)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3784..4372
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4273..4292
                     /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 cgcgttggtc ttgatcaatt gtcccagcag
      181 agttggtgca tgggaaagcc tttcttggcc ttatcagcag ctcagcctgg gaggttggcc
      241 cgggggctga ccaagcacaa aatagaaaag ccactaataa gatccagctg gaacaaatag
      301 cctctattta ttttaggaaa ctaatgatat gtcatcatga cacatatcta accttagtat
      361 ttctgattgg acaggccagt tagagctcgg gctgggcata aaagtcaggg cagagccatc
      421 tattgcttac atttgcttct gggatccaga tctttcgaag ctagcgccac catggtgagc
      481 aagggcgagg cagtgatcaa ggagttcatg cggttcaagg tgcacatgga gggctccatg
      541 aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag
      601 accgccaagc tgaaggtgac caagggtggc cccctgccct tctcctggga catcctgtcc
      661 cctcagttca tgtacggctc cagggccttc accaagcacc ccgccgacat ccccgactac
      721 tataagcagt ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc
      781 ggcgccgtga ccgtgaccca ggacacctcc ctggaggacg gcaccctgat ctacaaggtg
      841 aagctccgcg gcaccaactt ccctcctgac ggccccgtaa tgcagaagaa gacaatgggc
      901 tgggaagcgt ccaccgagcg gttgtacccc gaggacggcg tgctgaaggg cgacattaag
      961 atggccctgc gcctgaagga cggcggccgc tacctggcgg acttcaagac cacctacaag
     1021 gccaagaagc ccgtgcagat gcccggcgcc tacaacgtcg accgcaagtt ggacatcacc
     1081 tcccacaacg aggactacac cgtggtggaa cagtacgaac gctccgaggg ccgccactcc
     1141 accggcggca tggacgagct gtacaagtaa gtcgacggcg cgccgcggcc gcgaattcga
     1201 tatcataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     1261 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     1321 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttagttct tgccacggcg
     1381 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     1441 aattccgtgg ctcgagagat cttcgactgt gccttctagt tgccagccat ctgttgtttg
     1501 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     1561 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     1621 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgaga tctcacgtgc
     1681 ggaccgagcg gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc
     1741 gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg
     1801 cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc
     1861 ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg
     1921 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     1981 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     2041 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     2101 gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg
     2161 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     2221 ccaaactgga acaacactca accctatctc gggctattct tttgatttat aagggatttt
     2281 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     2341 taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc
     2401 cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     2461 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     2521 gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt
     2581 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     2641 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     2701 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     2761 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     2821 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     2881 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     2941 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     3001 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     3061 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     3121 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     3181 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     3241 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     3301 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     3361 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     3421 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     3481 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     3541 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     3601 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     3661 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     3721 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     3781 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     3841 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     3901 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     3961 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     4021 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     4081 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     4141 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     4201 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     4261 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     4321 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     4381 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgt
//
