LOCUS       sequence-462677-        4834 bp    DNA     circular SYN 15-APR-2025
DEFINITION  AiE0001m 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 4834)
  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 4834)
  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: AiP11272 - scAAV-AiE0001m-minBG-SYFP2-WPRE3-BG...
FEATURES             Location/Qualifiers
     source          1..4834
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      824..833
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(830..832,833..835,836..1546)
                     /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(875..896)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1136..1155)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1483..1504
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1639..1659)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1839..1950
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2189..2206)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2189..2205)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2198..2220)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2414..2433)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2533..2555
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2593..2611)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2679..2783
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2784..2852,2853..3644)
                     /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(3002..3021)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3815..4403
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4304..4323
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    4691..4712
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4727..4732,4733..4750,4751..4757)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4765..4781
                     /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     4770..4792
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4789..4805
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4789..4805
                     /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 gtccagtggc
      181 tagactgaag gaccctttgg aagggcttca gtttgcatta taaggaagga agtgcccagg
      241 cctatcattg ggataattaa tttgctatca ccgctctgtg ctgagaatgt aagcgcaaaa
      301 gaggagcaga acgtaatcgc tgacaaatta ctctgtgtgt actgaacatc agaatacatg
      361 cagctgccac ccagacaccc cccccccatt gtaccttagc agacaatagt tttcaactgg
      421 gactcagtac aatgagccac aaaagggaca tgaggtaagt gtagggtgta cgcagcagga
      481 cattttgatg aaatatctag gggacaattt aatatgaaat atcagctggt aaaagagacc
      541 agctgcttac taaggagaat gactcaagct ataactccag ctcctattat atataatgag
      601 attggcaaaa ttggactttt attctgggaa gaaaactgct gattttgaat gtgtcatggg
      661 tatgcctgca gactcccaac ttctaaggga atttgctttt ctgggttgaa ttaatactgt
      721 gaaccccagg gagctcgggc tgggcataaa agtcagggca gagccatcta ttgcttacat
      781 ttgcttctgg gatccagatc tttcgaagct agcgctaccg gtcgccacca tggtgagcaa
      841 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
      901 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
      961 cctgaagctg atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
     1021 cctgggctac ggcgtgcagt gcttcgcccg ctaccccgac cacatgaagc agcacgactt
     1081 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
     1141 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     1201 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
     1261 caactacaac agccacaacg tctatatcac cgccgacaag cagaagaacg gcatcaaggc
     1321 caacttcaag atccgccaca acatcgagga cggcggcgtg cagctcgccg accactacca
     1381 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcta
     1441 ccagtccaag ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     1501 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaag tcgacggcgc
     1561 gccgcggccg cgaattcgat atcataatca acctctggat tacaaaattt gtgaaagatt
     1621 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     1681 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     1741 gttagttctt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     1801 tcggctgttg ggcactgaca attccgtggc tcgagcgact gtgccttcta gttgccagcc
     1861 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     1921 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     1981 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatga
     2041 ctagtccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc
     2101 ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gagagggaca
     2161 gatccgggcc cgcatgcgtc gacaattcac tggccgtcgt tttacaacgt cgtgactggg
     2221 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc
     2281 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
     2341 aatggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatat
     2401 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc
     2461 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     2521 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     2581 cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
     2641 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     2701 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     2761 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     2821 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     2881 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     2941 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     3001 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     3061 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     3121 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     3181 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     3241 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     3301 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     3361 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     3421 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     3481 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     3541 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     3601 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     3661 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     3721 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     3781 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     3841 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     3901 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     3961 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     4021 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     4081 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     4141 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     4201 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     4261 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     4321 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     4381 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     4441 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     4501 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     4561 agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt
     4621 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
     4681 gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc
     4741 ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct
     4801 atgaccatga ttacgccaag ctctcgagat ctag
//
