LOCUS       sequence-470856-        4667 bp    DNA     circular SYN 30-APR-2025
DEFINITION  AiE0356h_3xC4 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 4667)
  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 4667)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 30, 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: AiP14287 -
            pAAV-AiE0356h_3xC4-minBG-SYFP2-WPRE...
FEATURES             Location/Qualifiers
     source          1..4667
                     /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      679..688
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(685..687,688..690,691..1401)
                     /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(730..751)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(991..1010)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1338..1359
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1494..1514)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1701..1812
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1930..2070
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1930..2059
                     /label=AAV2 ITR
     rep_origin      2145..2600
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2232..2251)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2442..2463
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2617..2636)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2736..2758
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2796..2814)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2882..2986
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2987..3055,3056..3847)
                     /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(3205..3224)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4018..4606
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4507..4526
                     /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 gcgtcctggc cccaatcccg ggctgtgact gaccttcaga
      181 tgcaggcagc agagagaaga ggacaaaagt gcaccgctgg ggaaaggcaa gcagctggca
      241 cccagggcac caaagacccg cacagtgagc atcaaggcca gcaagggcac acctggcccc
      301 aatcccgggc tgtgactgac cttcagatgc aggcagcaga gagaagagga caaaagtgca
      361 ccgctgggga aaggcaagca gctggcaccc agggcaccaa agacccgcac agtgagcatc
      421 aaggccagca agggcacacc tggccccaat cccgggctgt gactgacctt cagatgcagg
      481 cagcagagag aagaggacaa aagtgcaccg ctggggaaag gcaagcagct ggcacccagg
      541 gcaccaaaga cccgcacagt gagcatcaag gccagcaagg gcacagagct cgggctgggc
      601 ataaaagtca gggcagagcc atctattgct tacatttgct tctgggatcc agatctttcg
      661 aagctagcgc taccggtcgc caccatggtg agcaagggcg aggagctgtt caccggggtg
      721 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
      781 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agctgatctg caccaccggc
      841 aagctgcccg tgccctggcc caccctcgtg accaccctgg gctacggcgt gcagtgcttc
      901 gcccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
      961 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     1021 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     1081 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
     1141 atcaccgccg acaagcagaa gaacggcatc aaggccaact tcaagatccg ccacaacatc
     1201 gaggacggcg gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     1261 cccgtgctgc tgcccgacaa ccactacctg agctaccagt ccaagctgag caaagacccc
     1321 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     1381 ggcatggacg agctgtacaa gtaagtcgac ggcgcgccgc ggccgcgaat tcgatatcat
     1441 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     1501 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     1561 atggctttca ttttctcctc cttgtataaa tcctggttag ttcttgccac ggcggaactc
     1621 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc
     1681 gtggctcgag agatcttcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     1741 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     1801 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     1861 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gagatctcac gtgcggaccg
     1921 agcggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc
     1981 tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag
     2041 tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc
     2101 atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg
     2161 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     2221 cttagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     2281 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     2341 cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac
     2401 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     2461 tggaacaaca ctcaactcta tctcgggcta ttcttttgat ttataaggga ttttgccgat
     2521 ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     2581 aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg atgccgcata
     2641 gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     2701 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     2761 ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata
     2821 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     2881 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     2941 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     3001 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     3061 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     3121 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     3181 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     3241 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     3301 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     3361 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     3421 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     3481 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     3541 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     3601 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     3661 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     3721 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     3781 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     3841 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     3901 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     3961 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     4021 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     4081 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     4141 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     4201 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     4261 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     4321 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     4381 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     4441 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     4501 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     4561 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     4621 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgt
//
