LOCUS       sequence-461983-        4577 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2100m_C2 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 4577)
  AUTHORS   Ben-Simon Y, Hooper M, Narayan S, Daigle T, Dwivedi D, Way SW, Oster
            A, Stafford DA, Mich JK, Taormina MJ, Martinez RA, Opitz-Araya X, 
            Roth JR, Allen S, Ayala A, Bakken TE, Barcelli T, Barta S, Bendrick 
            J, Bertagnolli D, Bowlus J, Boyer G, Brouner K, Casian B, Casper T, 
            Chakka AB, Chakrabarty R, Chance RK, Chavan S, Departee M, Donadio 
            N, Dotson N, Egdorf T, Gabitto M, Garcia J, Gary A, Gasperini M, 
            Goldy J, Gore BB, Graybuck L, Greisman N, Haeseleer F, Halterman C, 
            Helback O, Hockemeyer D, Huang C, Huff S, Hunker A, Johansen N, 
            Juneau Z, Kalmbach B, Khem S, Kussick E, Kutsal R, Larsen R, Lee C, 
            Lee AY, Leibly M, Lenz GH, Liang E, Lusk N, Malone J, Mollenkopf T, 
            Morin E, Newman D, Ng L, Ngo K, Omstead V, Oyama A, Pham T, Pom CA, 
            Potekhina L, Ransford S, Rette D, Rimorin C, Rocha D, Ruiz A, 
            Sanchez REA, Sedeno-Cortes A, Sevigny JP, Shapovalova N, Shulga L, 
            Sigler AR, Siverts LA, Somasundaram S, Stewart K, Szelenyi E, Tieu 
            M, Trader C, van Velthoven CTJ, Walker M, 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 J, Tasic B
  TITLE     A suite of enhancer AAVs and transgenic mouse lines for genetic 
            access to cortical cell types (preprint).
  JOURNAL   bioRxiv [Preprint]. 2024 Sep 26:2024.06.10.597244. doi: 
            10.1101/2024.06.10.597244.
  PUBMED    38915722
REFERENCE   2  (bases 1 to 4577)
  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: "bioRxiv 
            [Preprint]. 2024 Sep 26:2024.06.10.597244. doi: 
            10.1101/2024.06.10.597244."
COMMENT     Sequence Label: AiP1325 - pAAV-AiE2100m_C2-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4577
                     /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      589..598
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(595..597,598..600,601..1311)
                     /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(640..661)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(901..920)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1248..1269
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1404..1424)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1611..1722
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1840..1980
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1840..1969
                     /label=AAV2 ITR
     rep_origin      2055..2510
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2142..2161)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2352..2373
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2527..2546)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2646..2668
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2706..2724)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2792..2896
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2897..2965,2966..3757)
                     /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(3115..3134)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3928..4516
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4417..4436
                     /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 gcgttaaagc aagaagggaa aatggatttc ttaatgagtc
      181 tgtatacagt gcagtatcta tgagctatgg tgtgtgtgac tgccaaggca gtcagatgct
      241 attcaagcaa ggaatgctgc tctgagatca gctagtacgt aagaaagaca tcctggctcc
      301 tggaagatga caagagtttg cctccgtagt atttgaaaga aagtggtcat ggccatcatg
      361 aatcaatcac ttcaggaaaa ggttgagcat gtgctgtgac gactgcagat ctttttatag
      421 ctgtggattc ttttatagct gctgagatac gacgatagga gtggagcggt agttatcgtt
      481 gatttgtttc ttaaggagct cgggctgggc ataaaagtca gggcagagcc atctattgct
      541 tacatttgct tctgggatcc agatctttcg aagctagcgc taccggtcgc caccatggtg
      601 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
      661 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
      721 ctgaccctga agctgatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
      781 accaccctgg gctacggcgt gcagtgcttc gcccgctacc ccgaccacat gaagcagcac
      841 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
      901 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
      961 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     1021 gagtacaact acaacagcca caacgtctat atcaccgccg acaagcagaa gaacggcatc
     1081 aaggccaact tcaagatccg ccacaacatc gaggacggcg gcgtgcagct cgccgaccac
     1141 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     1201 agctaccagt ccaagctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     1261 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaagtcgac
     1321 ggcgcgccgc ggccgcgaat tcgatatcat aatcaacctc tggattacaa aatttgtgaa
     1381 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     1441 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     1501 tcctggttag ttcttgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     1561 ggggctcggc tgttgggcac tgacaattcc gtggctcgag agatcttcga ctgtgccttc
     1621 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     1681 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     1741 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     1801 tagcaggcat gagatctcac gtgcggaccg agcggccgca ggaaccccta gtgatggagt
     1861 tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc
     1921 gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg
     1981 ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgtc
     2041 aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     2101 gcgcagcgtg accgctacac ttgccagcgc cttagcgccc gctcctttcg ctttcttccc
     2161 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     2221 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg
     2281 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     2341 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaactcta tctcgggcta
     2401 ttcttttgat ttataaggga ttttgccgat ttcggtctat tggttaaaaa atgagctgat
     2461 ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt tatggtgcac
     2521 tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc
     2581 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     2641 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg
     2701 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     2761 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     2821 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     2881 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     2941 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     3001 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     3061 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     3121 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     3181 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     3241 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     3301 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     3361 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     3421 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     3481 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     3541 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     3601 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     3661 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     3721 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     3781 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     3841 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     3901 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     3961 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     4021 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     4081 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     4141 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     4201 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     4261 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     4321 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     4381 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     4441 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     4501 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     4561 gccttttgct cacatgt
//
