LOCUS       sequence-462679-        4759 bp    DNA     circular SYN 15-APR-2025
DEFINITION  AiE0024m 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 4759)
  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 4759)
  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: AiP11281 - scAAV-AiE0024m-minBG-SYFP2-WPRE3-BG...
FEATURES             Location/Qualifiers
     source          1..4759
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      749..758
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(755..757,758..760,761..1471)
                     /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(800..821)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1061..1080)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1408..1429
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1564..1584)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1764..1875
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2114..2131)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2114..2130)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2123..2145)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2339..2358)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2458..2480
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2518..2536)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2604..2708
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2709..2777,2778..3569)
                     /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(2927..2946)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3740..4328
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4229..4248
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    4616..4637
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4652..4657,4658..4675,4676..4682)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4690..4706
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     4695..4717
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4714..4730
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4714..4730
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aaagcttccc ggggggatct gggccactcc ctctctgcgc gctcgctcgc tcactgaggc
       61 cgcccgggca aagcccgggc ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc
      121 aactccatca ctaggggttc ctggaggggt ggagtcgtga cctaggacgc gtctagcaaa
      181 agagctttgg ctctaacagc ttattttatc ctcaccccct aaaaacctcc aaaaagttct
      241 catttagcaa aaaattctac cagcaagtga cattttatga ttatctgatt ccacataaat
      301 taataattaa ctgctgaata tttgaggtct tatgttagcc ccctctggct tttaagaaac
      361 atatttcatt gaaaccaaat ccttccttca aatgctgccc agggaaagaa accaaagtca
      421 ccaatctggc atattcatga gaggtcaaaa tagtaaaacc agggccctga gttagtctga
      481 aattgcttag gggacgctgc aagaggagcc atgactgact aagagcagag cccatctgga
      541 ccctgaattc tgggaagctt ctggcaacag gcagagaaag agagaaaata gctctcatct
      601 gtgcagggat gatcatcagt tccagcatcc ctctgaacac agaaaattaa tgtaggagct
      661 cgggctgggc ataaaagtca gggcagagcc atctattgct tacatttgct tctgggatcc
      721 agatctttcg aagctagcgc taccggtcgc caccatggtg agcaagggcg aggagctgtt
      781 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
      841 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agctgatctg
      901 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctgg gctacggcgt
      961 gcagtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     1021 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
     1081 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     1141 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca
     1201 caacgtctat atcaccgccg acaagcagaa gaacggcatc aaggccaact tcaagatccg
     1261 ccacaacatc gaggacggcg gcgtgcagct cgccgaccac taccagcaga acacccccat
     1321 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agctaccagt ccaagctgag
     1381 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
     1441 gatcactctc ggcatggacg agctgtacaa gtaagtcgac ggcgcgccgc ggccgcgaat
     1501 tcgatatcat aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     1561 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     1621 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttag ttcttgccac
     1681 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     1741 tgacaattcc gtggctcgag cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     1801 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     1861 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     1921 gcaggacagc aagggggagg attgggaaga caatagcagg catgactagt ccactccctc
     1981 tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt
     2041 tgcccgggcg gcctcagtga gcgagcgagc gcgcagagag ggacagatcc gggcccgcat
     2101 gcgtcgacaa ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta
     2161 cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg
     2221 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg cgcctgatgc
     2281 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catatggtgc actctcagta
     2341 caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg
     2401 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     2461 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc
     2521 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag
     2581 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     2641 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     2701 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     2761 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     2821 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     2881 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     2941 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     3001 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     3061 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     3121 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     3181 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     3241 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     3301 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     3361 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     3421 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     3481 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     3541 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     3601 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     3661 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     3721 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     3781 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     3841 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     3901 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     3961 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     4021 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     4081 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     4141 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     4201 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     4261 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     4321 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     4381 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     4441 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     4501 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     4561 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
     4621 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
     4681 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg
     4741 ccaagctctc gagatctag
//
