LOCUS       sequence-438954-        4882 bp    DNA     circular SYN 15-APR-2025
DEFINITION  AiE2144m 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 4882)
  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 4882)
  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: AiP1432 - pAAV-AiE2144m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4882
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(6..25)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     216..237
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(391..410)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     510..532
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(570..588)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        656..760
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(761..829,830..1621)
                     /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(979..998)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1792..2380
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2281..2300
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2442..2571
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      3335..3344
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3341..3343,3344..3346,3347..4057)
                     /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(3386..3407)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3647..3666)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3994..4015
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4150..4170)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    4357..4468
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4586..4726
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4586..4715
                     /label=AAV2 ITR
     rep_origin      join(4801..4882,1..374)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
       61 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
      121 caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
      181 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
      241 aacactcaac tctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggt
      301 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
      361 aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag
      421 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
      481 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
      541 gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa
      601 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
      661 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
      721 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
      781 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
      841 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
      901 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
      961 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     1021 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     1081 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     1141 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     1201 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     1261 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     1321 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     1381 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     1441 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     1501 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     1561 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     1621 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     1681 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     1741 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     1801 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     1861 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     1921 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     1981 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     2041 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     2101 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     2161 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     2221 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     2281 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     2341 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     2401 tttacggttc ctggcctttt gctggccttt tgctcacatg tcctgcaggc agctgcgcgc
     2461 tcgctcgctc actgaggccg cccgggcgtc gggcgacctt tggtcgcccg gcctcagtga
     2521 gcgagcgagc gcgcagagag ggagtggcca actccatcac taggggttcc tgcggccgca
     2581 cgcgttgtaa gagtaataat cgccggcggg tgtcctgtgg gtgccttgag ggtaagaggc
     2641 tcaagaaatc ttcaaagtca gactccattc aagagctgaa ctaatgttgc ctgccactag
     2701 cgacattaac cctcagggag ggttggacag gatctgggga agctgcaggt gcagagtgca
     2761 ctagaatttg aaccccagga tacagacaag aagctgccta aaggcagaga gccctgggta
     2821 ccgtggtccc aggaggccaa agccccggag gtcacagagc tgagctcatt cttcctctcc
     2881 agctgcagat gttgtaaggg aaaaacaaca aaacaaacag gaagttctga ggatggtttc
     2941 ccacgtgttt ctctccatgt ttctctccag tcactgcctc gtcacccaaa ccagttttct
     3001 cctctcagcc tcttgtgctc ccagatcagg cctccctccc tcagcccaga tgtccccaac
     3061 gggggtgggg ggtggtttat gcttcgcttc taggatgcct gcatcttaga aaggcaaggt
     3121 ggcaggctgc acaggcaact gcattatcat ttcttaatgt catttttttc tgatccgtat
     3181 gatgggggaa atgtgctggg tgtggctcac tcctgtaatc gtagcactca agacgccggc
     3241 ggagctcggg ctgggcataa aagtcagggc agagccatct attgcttaca tttgcttctg
     3301 ggatccagat ctttcgaagc tagcgctacc ggtcgccacc atggtgagca agggcgagga
     3361 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
     3421 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagct
     3481 gatctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgggcta
     3541 cggcgtgcag tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     3601 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
     3661 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     3721 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
     3781 cagccacaac gtctatatca ccgccgacaa gcagaagaac ggcatcaagg ccaacttcaa
     3841 gatccgccac aacatcgagg acggcggcgt gcagctcgcc gaccactacc agcagaacac
     3901 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagct accagtccaa
     3961 gctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
     4021 cgccgggatc actctcggca tggacgagct gtacaagtaa gtcgacggcg cgccgcggcc
     4081 gcgaattcga tatcataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     4141 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     4201 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttagttct
     4261 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     4321 gggcactgac aattccgtgg ctcgagagat cttcgactgt gccttctagt tgccagccat
     4381 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     4441 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     4501 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgaga
     4561 tctcacgtgc ggaccgagcg gccgcaggaa cccctagtga tggagttggc cactccctct
     4621 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt
     4681 gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg
     4741 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt
     4801 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     4861 ctacacttgc cagcgcctta gc
//
