LOCUS       sequence-461982-        6323 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2104m 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 6323)
  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 6323)
  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: AiP1288 - pAAV-AiE2104m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..6323
                     /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      2335..2344
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2341..2343,2344..2346,2347..3057)
                     /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(2386..2407)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2647..2666)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2994..3015
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3150..3170)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    3357..3468
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3586..3726
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3586..3715
                     /label=AAV2 ITR
     rep_origin      3801..4256
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3888..3907)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4098..4119
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4273..4292)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4392..4414
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4452..4470)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4538..4642
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4643..4711,4712..5503)
                     /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(4861..4880)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5674..6262
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6163..6182
                     /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 gcgtcccaga aatcaaagac atagaaagat aattttctgt
      181 aaacaatgtt tcttattgtt ttgggccata ttccaatttc caaatgtaga tctattgaac
      241 aaagtttcac atcaattcaa atgtagtatt gaaatgtatt ttcagtggcg aaaagtgaga
      301 tatcatctta caatgagcat catgttcatt cctagaaata tctcagtgct gcagatattg
      361 agttcatact tctcagattc aaaattacat accttttatg aaaatgagaa ctaactattc
      421 aaacactatc ccatctgagg tcacatcttg gacttgagtc ccagaagcat ctattctgga
      481 atctggctga ttctactatt tgttaatagg gagactaaaa tagcaaaaat aagccatagg
      541 aaaagaaaat aaaggaaata ctgtgtcagc ccatctaaat gtattttctg tatcatccct
      601 tgtttctgaa agctgaggca tacagactca ggaatcaata atcaacatga attgccttta
      661 ttagacagaa gtctgcacta tgcagtgaga gcaattttcc tgatgaaggt tgtgtacatg
      721 gagctatatt gttcccacag ggtaaagtga tagggaaggt gcctactttg ttttaaatga
      781 ctaaagtcaa aatagagcaa acataaccca gtagtttgca gacagtgaat aagagagctg
      841 tttcttgtgg tccagagaaa aaagatgctg tttgtatatg ctcagcctag ggagtgtcac
      901 tattaggagg tatagccttg gtggaatagg tgtgtcattg caggtatggg ctttaagacc
      961 ctcgtcctag ctatctggaa gccagtcttc cattagcggc cttcagatga agatgtagaa
     1021 atctcagttc tccctgcatc gtgcctgcct agatgctgcc aagctcctat gttgatgata
     1081 atcaaataag cctctgaacc tgtaagccag ccccaattaa atgctgtcct ttttaagact
     1141 tgacttggtc atggtgtctg ttcacaggag taaagcccaa actaagacag aagatggtac
     1201 cagagtctgg ggtattgttg tgataggtct gaccatgctt ttgtttggaa gaatatggat
     1261 tggggacttt ggatttggaa agcagtggaa tcctttaagt agggcataat gggctatcct
     1321 agtagtaata tggaagactt tctcgctgag tatgatttga acactgcaga tctagtctaa
     1381 gtggtttcag tggagaagga ttccggtatg tggtgtagaa attatttttg caatattttg
     1441 gtgaagaatg tgactacttt ttgccattgg ccaaagagtc ttccagaggt gagggtgaag
     1501 agatttagat taattgcttt gaaaaagaaa gtctcaaaaa agcctagcag agactttgtt
     1561 gtctgattaa gtctcatgaa gagtgttttg aacaagtaga ccaagcttag aagtgaaaga
     1621 tataaaatat atagttggag tactaaaaga atgccaggaa ctggagctgg accctgtgtt
     1681 gaaagatatt aaattgaatt aagggagcgg tgagtttggg gcaagattcc cacccagcta
     1741 agtttaggtc taggcatggt agtaaaaacc tcccatccca agagtcaatg acaagcagat
     1801 ctctgagttc aaggcaagcc cagacagagc aagttctagg tgaagaaaaa cgtaaaccca
     1861 ggcttggtag accatacctt taatcccagt gcttagcaga ttggcttgca tttgtctgag
     1921 ttcaaagtca gtcttcagag caagttccag gacagccaag cttaggcagt ggaggagttg
     1981 gaaaacagaa aggctgatga tagtgtagta gaacaagggg accatgttcc agtcccagca
     2041 agcagcaaaa cttggcagct ttagacacat ggctctggtt ttagagtcta gaatagaagt
     2101 gactactggg ccaattagtg ctggttagct agagataaga aattagtggt gattaagaag
     2161 agaccagcat cagtgaggta aaatattctg gaaaatattt tctgagagca caatgaagct
     2221 gtgttccaga gatagtcaag ggagctcggg ctgggcataa aagtcagggc agagccatct
     2281 attgcttaca tttgcttctg ggatccagat ctttcgaagc tagcgctacc ggtcgccacc
     2341 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
     2401 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
     2461 ggcaagctga ccctgaagct gatctgcacc accggcaagc tgcccgtgcc ctggcccacc
     2521 ctcgtgacca ccctgggcta cggcgtgcag tgcttcgccc gctaccccga ccacatgaag
     2581 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
     2641 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     2701 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     2761 aagctggagt acaactacaa cagccacaac gtctatatca ccgccgacaa gcagaagaac
     2821 ggcatcaagg ccaacttcaa gatccgccac aacatcgagg acggcggcgt gcagctcgcc
     2881 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     2941 tacctgagct accagtccaa gctgagcaaa gaccccaacg agaagcgcga tcacatggtc
     3001 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa
     3061 gtcgacggcg cgccgcggcc gcgaattcga tatcataatc aacctctgga ttacaaaatt
     3121 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     3181 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     3241 tataaatcct ggttagttct tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     3301 tggacagggg ctcggctgtt gggcactgac aattccgtgg ctcgagagat cttcgactgt
     3361 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     3421 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     3481 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     3541 agacaatagc aggcatgaga tctcacgtgc ggaccgagcg gccgcaggaa cccctagtga
     3601 tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg
     3661 tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc
     3721 tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     3781 tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     3841 ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt
     3901 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     3961 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg
     4021 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     4081 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca actctatctc
     4141 gggctattct tttgatttat aagggatttt gccgatttcg gtctattggt taaaaaatga
     4201 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg
     4261 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
     4321 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     4381 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     4441 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     4501 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     4561 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     4621 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     4681 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     4741 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     4801 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     4861 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     4921 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     4981 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     5041 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     5101 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     5161 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     5221 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     5281 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     5341 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     5401 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     5461 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     5521 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     5581 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     5641 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     5701 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     5761 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     5821 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     5881 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     5941 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     6001 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     6061 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     6121 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     6181 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     6241 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     6301 ttgctggcct tttgctcaca tgt
//
