LOCUS       sequence-470847-        4883 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0807h_3xC2 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 4883)
  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 4883)
  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: AiP14276 -
            pAAV-AiE0807h_3xC2-minBG-SYFP2-WPRE...
FEATURES             Location/Qualifiers
     source          1..4883
                     /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      895..904
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(901..903,904..906,907..1617)
                     /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(946..967)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1207..1226)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1554..1575
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1710..1730)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1917..2028
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2146..2286
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2146..2275
                     /label=AAV2 ITR
     rep_origin      2361..2816
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2448..2467)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2658..2679
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2833..2852)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2952..2974
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3012..3030)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3098..3202
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3203..3271,3272..4063)
                     /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(3421..3440)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4234..4822
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4723..4742
                     /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 gcgtgtggaa ccctttcttg gtgcacactt tgtatttttt
      181 ttataaatct atgccattct gactcagcac tgattactac atgcacaatg atagcaatcc
      241 acaggcaagc attcatattg ctaatgatac tgctttgcag agtgattatc tgagagccat
      301 tctcagctgc tgcatcagga ttacagcatt tttgcaacca caaattaagg caggggtgaa
      361 aaagtggaac cctttcttgg tgcacacttt gtattttttt tataaatcta tgccattctg
      421 actcagcact gattactaca tgcacaatga tagcaatcca caggcaagca ttcatattgc
      481 taatgatact gctttgcaga gtgattatct gagagccatt ctcagctgct gcatcaggat
      541 tacagcattt ttgcaaccac aaattaaggc aggggtgaaa aagtggaacc ctttcttggt
      601 gcacactttg tatttttttt ataaatctat gccattctga ctcagcactg attactacat
      661 gcacaatgat agcaatccac aggcaagcat tcatattgct aatgatactg ctttgcagag
      721 tgattatctg agagccattc tcagctgctg catcaggatt acagcatttt tgcaaccaca
      781 aattaaggca ggggtgaaaa agagctcggg ctgggcataa aagtcagggc agagccatct
      841 attgcttaca tttgcttctg ggatccagat ctttcgaagc tagcgctacc ggtcgccacc
      901 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
      961 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
     1021 ggcaagctga ccctgaagct gatctgcacc accggcaagc tgcccgtgcc ctggcccacc
     1081 ctcgtgacca ccctgggcta cggcgtgcag tgcttcgccc gctaccccga ccacatgaag
     1141 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
     1201 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     1261 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     1321 aagctggagt acaactacaa cagccacaac gtctatatca ccgccgacaa gcagaagaac
     1381 ggcatcaagg ccaacttcaa gatccgccac aacatcgagg acggcggcgt gcagctcgcc
     1441 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     1501 tacctgagct accagtccaa gctgagcaaa gaccccaacg agaagcgcga tcacatggtc
     1561 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa
     1621 gtcgacggcg cgccgcggcc gcgaattcga tatcataatc aacctctgga ttacaaaatt
     1681 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     1741 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     1801 tataaatcct ggttagttct tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     1861 tggacagggg ctcggctgtt gggcactgac aattccgtgg ctcgagagat cttcgactgt
     1921 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     1981 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     2041 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     2101 agacaatagc aggcatgaga tctcacgtgc ggaccgagcg gccgcaggaa cccctagtga
     2161 tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg
     2221 tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc
     2281 tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     2341 tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     2401 ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt
     2461 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     2521 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg
     2581 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     2641 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca actctatctc
     2701 gggctattct tttgatttat aagggatttt gccgatttcg gtctattggt taaaaaatga
     2761 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg
     2821 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
     2881 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     2941 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     3001 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     3061 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     3121 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     3181 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     3241 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     3301 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     3361 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     3421 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     3481 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     3541 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     3601 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     3661 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     3721 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     3781 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     3841 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     3901 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     3961 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     4021 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     4081 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     4141 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     4201 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     4261 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     4321 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     4381 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     4441 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     4501 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     4561 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     4621 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     4681 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     4741 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     4801 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     4861 ttgctggcct tttgctcaca tgt
//
