LOCUS       sequence-461887-        5052 bp    DNA     circular SYN 15-APR-2025
DEFINITION  AiE0550h 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 5052)
  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.
  JOURNAL   bioRxiv [Preprint]. 2024 Sep 26:2024.06.10.597244. doi: 
            10.1101/2024.06.10.597244.
  PUBMED    38915722
REFERENCE   2  (bases 1 to 5052)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: AiP13653 -
            pAAV-AiE0550h-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5052
                     /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      518..527
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(524..526,527..529,530..1240)
                     /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(830..849)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1177..1198
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1250..1306
                     /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             1319..1384
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1415..1792
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(1879..1899)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2086..2197
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2315..2455
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2315..2444
                     /label=AAV2 ITR
     rep_origin      2530..2985
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2617..2636)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2827..2848
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3002..3021)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3121..3143
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3181..3199)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3267..3371
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3372..3440,3441..4232)
                     /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(3590..3609)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4403..4991
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4892..4911
                     /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 gcgtcagcag aaccactgcc tatttggacc aaaaggggta
      181 gaagcagtac taaatttaaa aaggccacgg gtccttaaaa ttctaccggt aggaagcagg
      241 ctgagaaaac cactcagctg caaacatatg ccacctttca cgaaaaagga aagatgactc
      301 agagtggaac caagaaacca aatggtacag tcaagagccc tggagaacta ttcccaggct
      361 ttgagaccta ctacaggagc acttgcctca acagacttca aattcctatt gacaggtgac
      421 tcctgagctc gggctgggca taaaagtcag ggcagagcca tctattgctt acatttgctt
      481 ctgggatcca gatctttcga agctagcgct accggtcgcc accatggtct ccaagggcga
      541 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgatg tcaacggcca
      601 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
      661 gctgatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctggg
      721 ctacggcgtg cagtgcttcg cccgctaccc cgaccacatg aagcagcacg acttcttcaa
      781 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaaag acgacggcaa
      841 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
      901 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
      961 caacagccac aacgtctata tcaccgccga caagcagaag aacggcatca aggccaactt
     1021 caagatccgc cacaacatcg aggacggcgg cgtgcagctc gccgaccact accagcagaa
     1081 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gctaccagtc
     1141 caagctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
     1201 cgccgccggg atcactctcg gcatggacga gctgtacaag ggctctggtg ctaccaactt
     1261 ctcactgttg aaacaggcag gggatgtaga ggagaatcca gggcctggtg ctagtggaga
     1321 ctacaaagac catgacggag attataaaga tcatgacatc gattacaagg atgacgatga
     1381 caagtccgga ctcagatctg gaggctccgg aggcccagag ccagcgaagt ctgctcccgc
     1441 cccgaaaaag ggctccaaga aggcggtgac taaggcgcag aagaaaggcg gcaagaagcg
     1501 caagcgcagc cgcaaggaga gctattccat ctatgtgtac aaggttctga agcaggtcca
     1561 ccctgacacc ggcatttcgt ccaaggccat gggcatcatg aattcgtttg tgaacgacat
     1621 tttcgagcgc atcgcaggag aggcttcccg cctggcgcat tacaacaagc gctcgaccat
     1681 cacctcccgg gagatccaga cggccgtgcg cctgctgctg cctggggagt tggccaagca
     1741 cgccgtgtcc gagggtacta aggccatcac caagtacacc agcgctaagt aatgaggcgc
     1801 gccgcggccg cgaattcgat atcataatca acctctggat tacaaaattt gtgaaagatt
     1861 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     1921 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     1981 gttagttctt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     2041 tcggctgttg ggcactgaca attccgtggc tcgagagatc ttcgactgtg ccttctagtt
     2101 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     2161 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     2221 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     2281 ggcatgagat ctcacgtgcg gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc
     2341 actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc
     2401 ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc
     2461 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc
     2521 aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     2581 gcgtgaccgc tacacttgcc agcgccttag cgcccgctcc tttcgctttc ttcccttcct
     2641 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     2701 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac
     2761 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     2821 ttaatagtgg actcttgttc caaactggaa caacactcaa ctctatctcg ggctattctt
     2881 ttgatttata agggattttg ccgatttcgg tctattggtt aaaaaatgag ctgatttaac
     2941 aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca
     3001 gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg
     3061 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     3121 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg
     3181 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     3241 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     3301 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     3361 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     3421 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     3481 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     3541 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     3601 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     3661 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     3721 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     3781 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     3841 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     3901 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     3961 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     4021 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     4081 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     4141 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     4201 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     4261 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     4321 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     4381 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     4441 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     4501 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     4561 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     4621 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     4681 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     4741 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     4801 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     4861 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     4921 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     4981 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     5041 ttgctcacat gt
//
