LOCUS       sequence-461902-        5184 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2004m_6xC2 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 5184)
  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 5184)
  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: AiP12285 -
            pAAV-AiE2004m_6xC2-minCMV-SYFP2-WPR...
FEATURES             Location/Qualifiers
     source          1..5184
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        1096..1134
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     regulatory      1196..1205
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1202..1204,1205..1207,1208..1918)
                     /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(1247..1268)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1508..1527)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1855..1876
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2011..2031)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2218..2329
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2447..2587
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2447..2576
                     /label=AAV2 ITR
     rep_origin      2662..3117
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2749..2768)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2959..2980
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3134..3153)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3253..3275
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3313..3331)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3399..3503
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3504..3572,3573..4364)
                     /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(3722..3741)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4535..5123
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5024..5043
                     /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 gcgtttaccc tgttgaatat cactgactca ctaacttgca
      181 ttgccatgct aacttgcttt cagagagatc tcagaacaca tcatcttctg ctatttcaat
      241 acatgcacat taatttccta tcaacgtgtg ctgatcagga actctgtaat ctggcaccgt
      301 taccctgttg aatatcactg actcactaac ttgcattgcc atgctaactt gctttcagag
      361 agatctcaga acacatcatc ttctgctatt tcaatacatg cacattaatt tcctatcaac
      421 gtgtgctgat caggaactct gtaatctggc accgttaccc tgttgaatat cactgactca
      481 ctaacttgca ttgccatgct aacttgcttt cagagagatc tcagaacaca tcatcttctg
      541 ctatttcaat acatgcacat taatttccta tcaacgtgtg ctgatcagga actctgtaat
      601 ctggcaccgc ttaagttacc ctgttgaata tcactgactc actaacttgc attgccatgc
      661 taacttgctt tcagagagat ctcagaacac atcatcttct gctatttcaa tacatgcaca
      721 ttaatttcct atcaacgtgt gctgatcagg aactctgtaa tctggcaccg ttaccctgtt
      781 gaatatcact gactcactaa cttgcattgc catgctaact tgctttcaga gagatctcag
      841 aacacatcat cttctgctat ttcaatacat gcacattaat ttcctatcaa cgtgtgctga
      901 tcaggaactc tgtaatctgg caccgttacc ctgttgaata tcactgactc actaacttgc
      961 attgccatgc taacttgctt tcagagagat ctcagaacac atcatcttct gctatttcaa
     1021 tacatgcaca ttaatttcct atcaacgtgt gctgatcagg aactctgtaa tctggcaccg
     1081 cttaaggagc tcagaggtag gcgtgtacgg tgggaggcct atataagcag agctggttta
     1141 gtgaaccgtc agatcgcctg gggatccaga tctttcgaag ctagcgctac cggtcgccac
     1201 catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
     1261 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
     1321 cggcaagctg accctgaagc tgatctgcac caccggcaag ctgcccgtgc cctggcccac
     1381 cctcgtgacc accctgggct acggcgtgca gtgcttcgcc cgctaccccg accacatgaa
     1441 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
     1501 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     1561 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     1621 caagctggag tacaactaca acagccacaa cgtctatatc accgccgaca agcagaagaa
     1681 cggcatcaag gccaacttca agatccgcca caacatcgag gacggcggcg tgcagctcgc
     1741 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     1801 ctacctgagc taccagtcca agctgagcaa agaccccaac gagaagcgcg atcacatggt
     1861 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta
     1921 agtcgacggc gcgccgcggc cgcgaattcg atatcataat caacctctgg attacaaaat
     1981 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     2041 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     2101 gtataaatcc tggttagttc ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     2161 ctggacaggg gctcggctgt tgggcactga caattccgtg gctcgagaga tcttcgactg
     2221 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     2281 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     2341 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     2401 aagacaatag caggcatgag atctcacgtg cggaccgagc ggccgcagga acccctagtg
     2461 atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag
     2521 gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc
     2581 ctgcaggggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     2641 atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     2701 tggttacgcg cagcgtgacc gctacacttg ccagcgcctt agcgcccgct cctttcgctt
     2761 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     2821 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg
     2881 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     2941 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aactctatct
     3001 cgggctattc ttttgattta taagggattt tgccgatttc ggtctattgg ttaaaaaatg
     3061 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat
     3121 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc
     3181 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     3241 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     3301 cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
     3361 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     3421 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     3481 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     3541 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     3601 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     3661 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     3721 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     3781 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     3841 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     3901 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     3961 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     4021 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     4081 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     4141 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     4201 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     4261 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     4321 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     4381 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     4441 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     4501 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     4561 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     4621 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     4681 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     4741 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     4801 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     4861 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     4921 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     4981 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     5041 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     5101 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     5161 tttgctggcc ttttgctcac atgt
//
