LOCUS       sequence-471604-        4693 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE3103m 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 4693)
  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 4693)
  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: AiP20055 - pAAV-AiE3103m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4693
                     /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      705..714
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(711..713,714..716,717..1427)
                     /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(756..777)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1017..1036)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1364..1385
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1520..1540)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1727..1838
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1956..2096
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1956..2085
                     /label=AAV2 ITR
     rep_origin      2171..2626
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2258..2277)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2468..2489
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2643..2662)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2762..2784
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2822..2840)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2908..3012
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3013..3081,3082..3873)
                     /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(3231..3250)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4044..4632
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4533..4552
                     /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 gcgtggctgg ggagcataga gtaatgttaa tttagtcttc
      181 aacccttaag agaaacctta gacaaaggga aatgaacaaa tgtaggagga gcatccttct
      241 gtgggttgtc agttcacaga acacagtttt agaggacttt tggcttctct ccatctgact
      301 gaggagaacc aggtctcatt tgccaggtac tagtttttgt gcatccttta ggactcattt
      361 gctaatctgt gcaggagact cctcatgctc atatcacatg ggccaggggc cactgcagaa
      421 tgccagctct gctggaactg tttcttaggc tctcaggcag cccaggatgg gtgcagtggg
      481 aaaccaccag aggcagtgca cagagcccac aggagcagag caaacgtaat tagggggata
      541 tttctgccag agccccaaat ggcatgctgt tctgacctgt caaactacct agctctgtag
      601 tttccgccgc ggagctcggg ctgggcataa aagtcagggc agagccatct attgcttaca
      661 tttgcttctg ggatccagat ctttcgaagc tagcgctacc ggtcgccacc atggtgagca
      721 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
      781 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
      841 ccctgaagct gatctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
      901 ccctgggcta cggcgtgcag tgcttcgccc gctaccccga ccacatgaag cagcacgact
      961 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
     1021 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     1081 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     1141 acaactacaa cagccacaac gtctatatca ccgccgacaa gcagaagaac ggcatcaagg
     1201 ccaacttcaa gatccgccac aacatcgagg acggcggcgt gcagctcgcc gaccactacc
     1261 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagct
     1321 accagtccaa gctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     1381 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa gtcgacggcg
     1441 cgccgcggcc gcgaattcga tatcataatc aacctctgga ttacaaaatt tgtgaaagat
     1501 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     1561 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     1621 ggttagttct tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
     1681 ctcggctgtt gggcactgac aattccgtgg ctcgagagat cttcgactgt gccttctagt
     1741 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     1801 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     1861 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     1921 aggcatgaga tctcacgtgc ggaccgagcg gccgcaggaa cccctagtga tggagttggc
     1981 cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg
     2041 cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg
     2101 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag
     2161 caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     2221 agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc
     2281 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     2341 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca
     2401 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     2461 tttaatagtg gactcttgtt ccaaactgga acaacactca actctatctc gggctattct
     2521 tttgatttat aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa
     2581 caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc
     2641 agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct
     2701 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     2761 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag
     2821 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     2881 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     2941 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     3001 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     3061 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     3121 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     3181 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     3241 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     3301 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     3361 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     3421 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     3481 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     3541 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     3601 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     3661 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     3721 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     3781 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     3841 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     3901 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     3961 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     4021 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     4081 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     4141 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     4201 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     4261 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     4321 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     4381 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     4441 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     4501 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     4561 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     4621 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     4681 tttgctcaca tgt
//
