LOCUS       sequence-466278-        4758 bp    DNA     circular SYN 15-APR-2025
DEFINITION  AiE2283m 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 4758)
  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 4758)
  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: AiP1616 - pAAV-AiE2283m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4758
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      273..282
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(279..281,282..284,285..995)
                     /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(324..345)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(585..604)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     932..953
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1088..1108)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1295..1406
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1524..1664
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1524..1653
                     /label=AAV2 ITR
     rep_origin      1739..2194
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1826..1845)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2036..2057
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2211..2230)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2330..2352
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2390..2408)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2476..2580
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2581..2649,2650..3441)
                     /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(2799..2818)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3612..4200
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4101..4120
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   4262..4391
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 cttgctcact tcaccttctt ttgaaaatgt gcaggctagt tacaatgtgc accccgtcct
       61 tgccttttta ttgtttagtc aactcttaac cattgctttt atttaaatct tccctaaacc
      121 agtgatagag ttagggaaaa tattcataaa taaacaaaat aatgaaatct acgccggcgg
      181 agctcgggct gggcataaaa gtcagggcag agccatctat tgcttacatt tgcttctggg
      241 atccagatct ttcgaagcta gcgctaccgg tcgccaccat ggtgagcaag ggcgaggagc
      301 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
      361 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagctga
      421 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgggctacg
      481 gcgtgcagtg cttcgcccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
      541 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
      601 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
      661 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca
      721 gccacaacgt ctatatcacc gccgacaagc agaagaacgg catcaaggcc aacttcaaga
      781 tccgccacaa catcgaggac ggcggcgtgc agctcgccga ccactaccag cagaacaccc
      841 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagctac cagtccaagc
      901 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
      961 ccgggatcac tctcggcatg gacgagctgt acaagtaagt cgacggcgcg ccgcggccgc
     1021 gaattcgata tcataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc
     1081 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
     1141 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttagttcttg
     1201 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg
     1261 gcactgacaa ttccgtggct cgagagatct tcgactgtgc cttctagttg ccagccatct
     1321 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     1381 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     1441 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgagatc
     1501 tcacgtgcgg accgagcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct
     1561 gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc
     1621 ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc tgatgcggta
     1681 ttttctcctt acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac
     1741 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     1801 acacttgcca gcgccttagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     1861 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     1921 gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca
     1981 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     2041 ctcttgttcc aaactggaac aacactcaac tctatctcgg gctattcttt tgatttataa
     2101 gggattttgc cgatttcggt ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     2161 gcgaatttta acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc
     2221 tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga
     2281 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     2341 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata
     2401 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact
     2461 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     2521 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     2581 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     2641 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     2701 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     2761 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     2821 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     2881 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     2941 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     3001 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     3061 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     3121 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     3181 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     3241 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     3301 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     3361 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     3421 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     3481 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     3541 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     3601 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     3661 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     3721 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     3781 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     3841 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     3901 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     3961 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     4021 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     4081 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     4141 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     4201 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     4261 tcctgcaggc agctgcgcgc tcgctcgctc actgaggccg cccgggcgtc gggcgacctt
     4321 tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca actccatcac
     4381 taggggttcc tgcggccgca cgcgttgtaa gagtaataat cgccggcgaa gactgaagat
     4441 ccaatccaat gtaatcccca tctggaaatc cttgttccct ctctcccgag atgtgccccc
     4501 actattgtga caggagtggc cctgctacct tctctgggtc ataccatcca tgctgggaat
     4561 gcattatcca gctctgctcc catcatgcat cgttctttgt cttagcctga agaggaactg
     4621 ccttgtctgc tcttcttgta caaggaaaat cacatgactg cctctgcttc cctttttact
     4681 ttggtcacca ggaagcccat ggtcaaatgt gtggcctaat caaagaattg ttaggtcagc
     4741 agtttagcct tagggtca
//
