LOCUS       sequence-466252-        4670 bp DNA     circular SYN 24-MAR-2025
DEFINITION  AiE2158m 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 4670)
  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 4670)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Mar 24, 2025 from SnapGene Server 7.0.3
            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."
FEATURES             Location/Qualifiers
     source          1..4670
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     161..180
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   322..451
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      1003..1012
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1009..1725
                     /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(1054..1075)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1315..1334)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1662..1683
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1818..1838)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2025..2136
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2254..2394
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2254..2383
                     /label=AAV2 ITR
     rep_origin      2469..2924
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2556..2575)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2766..2787
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2941..2960)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3060..3082
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3120..3138)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3206..3310
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3311..4171
                     /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(3529..3548)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(4342..4670,1..260)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
       61 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
      121 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
      181 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
      241 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
      301 gctggccttt tgctcacatg tcctgcaggc agctgcgcgc tcgctcgctc actgaggccg
      361 cccgggcgtc gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag
      421 ggagtggcca actccatcac taggggttcc tgcggccgca cgcgtgttgg tgggagtata
      481 tgccatatag atacaggaag gctggggttg tcttaacata aacacagctt aaaggaacta
      541 ttctcaaatc tctctccttt ctgaccagct acctaagtca tcaagacaaa ctcttcaact
      601 ggcagaaagc tggcctagcc tcactatgcc tcccagcaca ctgccttctg gggcagcaag
      661 tgcctgggaa catcgtagaa gcacattccc aggacctgct gggctaaggc ttcacctcac
      721 atggtcacca tcaggccggg ttcaggcccc tcggggctct gcttttagca gcaatgggaa
      781 agaggcacag ggtaagagag tcagagggcg ataggcaggt ctgcaaagca aacactcaga
      841 aaattagaaa tagctcaaga aacaaatgcc ctgagaaagt ataaggctca ctgtaattct
      901 agcccttcag agctcgggct gggcataaaa gtcagggcag agccatctat tgcttacatt
      961 tgcttctggg atccagatct ttcgaagcta gcgctaccgg tcgccaccat ggtgagcaag
     1021 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
     1081 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
     1141 ctgaagctga tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     1201 ctgggctacg gcgtgcagtg cttcgcccgc taccccgacc acatgaagca gcacgacttc
     1261 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
     1321 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     1381 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     1441 aactacaaca gccacaacgt ctatatcacc gccgacaagc agaagaacgg catcaaggcc
     1501 aacttcaaga tccgccacaa catcgaggac ggcggcgtgc agctcgccga ccactaccag
     1561 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagctac
     1621 cagtccaagc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     1681 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaagt cgacggcgcg
     1741 ccgcggccgc gaattcgata tcataatcaa cctctggatt acaaaatttg tgaaagattg
     1801 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     1861 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     1921 ttagttcttg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
     1981 cggctgttgg gcactgacaa ttccgtggct cgagagatct tcgactgtgc cttctagttg
     2041 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     2101 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     2161 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
     2221 gcatgagatc tcacgtgcgg accgagcggc cgcaggaacc cctagtgatg gagttggcca
     2281 ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc
     2341 cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc
     2401 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata cgtcaaagca
     2461 accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     2521 cgtgaccgct acacttgcca gcgccttagc gcccgctcct ttcgctttct tcccttcctt
     2581 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     2641 ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg
     2701 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     2761 taatagtgga ctcttgttcc aaactggaac aacactcaac tctatctcgg gctattcttt
     2821 tgatttataa gggattttgc cgatttcggt ctattggtta aaaaatgagc tgatttaaca
     2881 aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt gcactctcag
     2941 tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga
     3001 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     3061 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg
     3121 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     3181 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     3241 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     3301 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     3361 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     3421 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     3481 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     3541 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     3601 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     3661 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     3721 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     3781 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     3841 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     3901 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     3961 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     4021 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     4081 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     4141 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     4201 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     4261 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     4321 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     4381 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     4441 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     4501 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     4561 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     4621 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
//
