LOCUS       sequence-461346-        4824 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0700m is an enhancer sequence, designed to drive AAV-mediated 
            transgene expression in specific brain cell types.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4824)
  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 4824)
  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: AiP12860 - pAAV-AiE0700m-minBG-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..4824
                     /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      836..845
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(842..844,845..847,848..1558)
                     /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(887..908)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1148..1167)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1495..1516
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1651..1671)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1858..1969
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2087..2227
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2087..2216
                     /label=AAV2 ITR
     rep_origin      2302..2757
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2389..2408)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2599..2620
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2774..2793)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2893..2915
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2953..2971)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3039..3143
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3144..3212,3213..4004)
                     /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(3362..3381)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4175..4763
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4664..4683
                     /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 gcgtttggct ttccttccaa gttgtatttt ggtgaagttt
      181 gactcctgga atctcacttc tgcactttag attagcatac tttaaaaagg aacattacag
      241 tacaatggaa aagtaaagat ctgggtttta tctcagcttt agttagtatt tataggaagc
      301 atggattaca taaagagatg acattatttc agtctgaaat tgatttgtac tatacgacat
      361 ttcattttgt ttgtgcacat tgattagata attcctcaaa tcagtttgaa aatttcatct
      421 ctgttcatag ctgaagataa ccaccaaatt gcttttaact aatttacata ataagtatag
      481 tagaggttta ttctatataa agtatcaaat ataaatattc atctcaaatt ctaaaggatt
      541 tagcaatgta aattacttat gaattacata cacatttgta tttaaatctt ttttcacttt
      601 tattaaaaat agatcctttt ctcatacaat atatcctgat tgcagttccc tcccctctac
      661 tcctcccagt tccttcccac ctccctttcc atctggatct ctctcctatt aaaggttgga
      721 cagaagagat ggcactgcat gtgagctcgg gctgggcata aaagtcaggg cagagccatc
      781 tattgcttac atttgcttct gggatccaga tctttcgaag ctagcgctac cggtcgccac
      841 catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
      901 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
      961 cggcaagctg accctgaagc tgatctgcac caccggcaag ctgcccgtgc cctggcccac
     1021 cctcgtgacc accctgggct acggcgtgca gtgcttcgcc cgctaccccg accacatgaa
     1081 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
     1141 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     1201 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     1261 caagctggag tacaactaca acagccacaa cgtctatatc accgccgaca agcagaagaa
     1321 cggcatcaag gccaacttca agatccgcca caacatcgag gacggcggcg tgcagctcgc
     1381 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     1441 ctacctgagc taccagtcca agctgagcaa agaccccaac gagaagcgcg atcacatggt
     1501 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta
     1561 agtcgacggc gcgccgcggc cgcgaattcg atatcataat caacctctgg attacaaaat
     1621 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     1681 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     1741 gtataaatcc tggttagttc ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     1801 ctggacaggg gctcggctgt tgggcactga caattccgtg gctcgagaga tcttcgactg
     1861 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     1921 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     1981 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     2041 aagacaatag caggcatgag atctcacgtg cggaccgagc ggccgcagga acccctagtg
     2101 atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag
     2161 gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc
     2221 ctgcaggggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     2281 atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     2341 tggttacgcg cagcgtgacc gctacacttg ccagcgcctt agcgcccgct cctttcgctt
     2401 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     2461 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg
     2521 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     2581 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aactctatct
     2641 cgggctattc ttttgattta taagggattt tgccgatttc ggtctattgg ttaaaaaatg
     2701 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat
     2761 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc
     2821 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     2881 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     2941 cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
     3001 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     3061 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     3121 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     3181 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     3241 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     3301 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     3361 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     3421 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     3481 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     3541 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     3601 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     3661 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     3721 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     3781 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     3841 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     3901 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     3961 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     4021 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     4081 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     4141 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     4201 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     4261 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     4321 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     4381 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     4441 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     4501 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     4561 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     4621 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     4681 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     4741 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     4801 tttgctggcc ttttgctcac atgt
//
