LOCUS       sequence-469797-        4993 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ExE0639m 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 4993)
  AUTHORS   Ben-Simon Y, Hooper M, Narayan S, Daigle TL, Dwivedi D, Way SW, 
            Oster A, Stafford DA, Mich JK, Taormina MJ, Martinez RA, Opitz-Araya
            X, Roth JR, Alexander JR, Allen S, Amster A, Arbuckle J, Ayala A, 
            Baker PM, Bakken TE, Barcelli T, Barta S, Bendrick J, Bertagnolli D,
            Bielstein C, Bishwakarma P, Bowlus J, Boyer G, Brouner K, Casian B, 
            Casper T, Chakka AB, Chakrabarty R, Chance RK, Chavan S, Clark M, 
            Colbert K, Collman F, Daniel S, Departee M, DiValentin P, Donadio N,
            Dotson N, Egdorf T, Fliss T, Gabitto M, Garcia J, Gary A, Gasperini 
            M, Gloe J, Goldy J, Gore BB, Graybuck L, Greisman N, Haeseleer F, 
            Halterman C, Haradon Z, Hastings SD, Helback O, Ho W, Hockemeyer D, 
            Huang C, Huff S, Hunker A, Johansen N, Jones D, Juneau Z, Kalmbach 
            B, Kannan M, Khem S, Kussick E, Kutsal R, Larsen R, Lee C, Lee AY, 
            Leibly M, Lenz GH, Li S, Liang E, Lusk N, Madigan Z, Malloy J, 
            Malone J, McCue R, Melchor J, Mollenkopf T, Moosman S, Morin E, 
            Newman D, Ng L, Ngo K, Omstead V, Otto S, Oyama A, Pena N, Pham T, 
            Phillips E, Pom CA, Potekhina L, Ransford S, Ray PL, Rette D, 
            Reynoldson C, Rimorin C, Rocha D, Ruiz A, Sanchez REA, Sawyer L, 
            Sedeno-Cortes A, Sevigny JP, Shapovalova N, Shepard N, Shulga L, 
            Sigler AR, Siverts L, Soliman S, Somasundaram S, Staats B, Stewart 
            K, Szelenyi E, Tieu M, Trader C, Tran A, van Velthoven CTJ, Walker 
            M, Wang Y, 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 JT, Tasic B
  TITLE     A suite of enhancer AAVs and transgenic mouse lines for genetic 
            access to cortical cell types.
  JOURNAL   Cell. 2025 May 16:S0092-8674(25)00513-6. doi: 
            10.1016/j.cell.2025.05.002.
  PUBMED    40403729
REFERENCE   2  (bases 1 to 4993)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell. 2025 
            May 16:S0092-8674(25)00513-6. doi: 10.1016/j.cell.2025.05.002."
COMMENT     Sequence Label: AiP12725 - pAAV-ExE0639m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4993
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      1005..1014
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1011..1013,1014..1016,1017..1727)
                     /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(1056..1077)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1317..1336)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1664..1685
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1820..1840)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2027..2138
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2256..2396
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2256..2385
                     /label=AAV2 ITR
     rep_origin      2471..2926
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2558..2577)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2768..2789
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2943..2962)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3062..3084
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3122..3140)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3208..3312
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3313..3381,3382..4173)
                     /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(3531..3550)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4344..4932
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4833..4852
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtaaagtg agttccagga cagccaggac tacacagaga
      181 aaccctgtct caaaaaaaca aaatcaaaac aaaacaaaca aacaaaaaag ctaatgactc
      241 catcatgact gtaacaaaca catcagtgcg gcagtgagag cccgtctgtc agcatcagca
      301 acagcattag tcagactgta tttgtgagca tatttgctta ggtctcttct aaataccctt
      361 cacttttctc tcagagaaac ccagttcatc gtattctgaa aaggagcggc cgtaaaggac
      421 tgatcctgtc tgaagcactt tggtataaaa gttgcttagc agtggggcag aaaagaaaaa
      481 aagcaattaa gtttatattt agtgatctat ctatacacat ctggagcaca tttgggaaag
      541 aattcaaaag ggccaattca ttgcatgcct cctgctacag aacgagtgtg ggagtcaagc
      601 tgcgatttcc acagcatcag acatttattg ttgacttcaa aaagttctcc cacttatgtg
      661 taattactat cctagcaaat ggctctgaaa tttcagcttc ttaagcataa ggcagagtgg
      721 tcctttaaaa gtaaaataaa acgtaggccc tatgagataa aattaagata aattaagaat
      781 cagttacttc caagacgaag cacttatggt gcatgccttc ttatataaag cagatcctta
      841 ccatgtatgt gtgctgtttg cttgccaaga ccaagatgtc ttccataact gtggcagcac
      901 tggactttga agagctcggg ctgggcataa aagtcagggc agagccatct attgcttaca
      961 tttgcttctg ggatccagat ctttcgaagc tagcgctacc ggtcgccacc atggtgagca
     1021 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
     1081 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
     1141 ccctgaagct gatctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     1201 ccctgggcta cggcgtgcag tgcttcgccc gctaccccga ccacatgaag cagcacgact
     1261 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
     1321 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     1381 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     1441 acaactacaa cagccacaac gtctatatca ccgccgacaa gcagaagaac ggcatcaagg
     1501 ccaacttcaa gatccgccac aacatcgagg acggcggcgt gcagctcgcc gaccactacc
     1561 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagct
     1621 accagtccaa gctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     1681 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa gtcgacggcg
     1741 cgccgcggcc gcgaattcga tatcataatc aacctctgga ttacaaaatt tgtgaaagat
     1801 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     1861 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     1921 ggttagttct tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
     1981 ctcggctgtt gggcactgac aattccgtgg ctcgagagat cttcgactgt gccttctagt
     2041 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     2101 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     2161 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     2221 aggcatgaga tctcacgtgc ggaccgagcg gccgcaggaa cccctagtga tggagttggc
     2281 cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg
     2341 cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg
     2401 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag
     2461 caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     2521 agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc
     2581 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     2641 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca
     2701 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     2761 tttaatagtg gactcttgtt ccaaactgga acaacactca actctatctc gggctattct
     2821 tttgatttat aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa
     2881 caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc
     2941 agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct
     3001 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     3061 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag
     3121 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     3181 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     3241 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     3301 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     3361 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     3421 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     3481 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     3541 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     3601 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     3661 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     3721 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     3781 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     3841 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     3901 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     3961 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     4021 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     4081 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     4141 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     4201 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     4261 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     4321 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     4381 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     4441 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     4501 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     4561 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     4621 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     4681 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     4741 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     4801 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     4861 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     4921 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     4981 tttgctcaca tgt
//
