LOCUS       sequence-466360-        4563 bp DNA     circular SYN 24-MAR-2025
DEFINITION  AiE3097m 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 4563)
  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.
  JOURNAL   bioRxiv [Preprint]. 2024 Sep 26:2024.06.10.597244. doi: 
            10.1101/2024.06.10.597244.
  PUBMED    38915722
REFERENCE   2  (bases 1 to 4563)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Mar 24, 2025 from SnapGene Server 7.0.3
            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."
FEATURES             Location/Qualifiers
     source          1..4563
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     415..434
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   576..705
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      1150..1159
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1156..1872
                     /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(1201..1222)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1462..1481)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1809..1830
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1965..1985)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2172..2283
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2401..2541
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2401..2530
                     /label=AAV2 ITR
     rep_origin      2616..3071
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2703..2722)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2913..2934
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3088..3107)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3207..3229
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3267..3285)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3353..3457
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3458..4318
                     /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(3676..3695)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(4489..4563,1..514)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
       61 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
      121 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
      181 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
      241 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
      301 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
      361 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
      421 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
      481 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
      541 gttcctggcc ttttgctggc cttttgctca catgtcctgc aggcagctgc gcgctcgctc
      601 gctcactgag gccgcccggg cgtcgggcga cctttggtcg cccggcctca gtgagcgagc
      661 gagcgcgcag agagggagtg gccaactcca tcactagggg ttcctgcggc cgcacgcgtt
      721 tgagggaaga gcagattctc agactaggga cattcaaatc ctaggtctct gacagttatt
      781 cacagacaat ggcagtttcc actggaaaca cagcagtcag caaatctctg cgacatcacc
      841 ctctgtcaca tactccctgg ataatctatt gtaggacctg cttcatgaca gctgagccaa
      901 gcctgagttt ttggtgaaca tcaagtttgc aaaaatttgg caacagttgt aagtttgctg
      961 agggcagatc cagttaaagt gcactgtggg ctgcacggga ctttagaatc acaggaccta
     1021 cgtggttctg tgctcagtaa ctgagtgacg ccggcggagc tcgggctggg cataaaagtc
     1081 agggcagagc catctattgc ttacatttgc ttctgggatc cagatctttc gaagctagcg
     1141 ctaccggtcg ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     1201 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
     1261 ggcgatgcca cctacggcaa gctgaccctg aagctgatct gcaccaccgg caagctgccc
     1321 gtgccctggc ccaccctcgt gaccaccctg ggctacggcg tgcagtgctt cgcccgctac
     1381 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     1441 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     1501 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     1561 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcaccgcc
     1621 gacaagcaga agaacggcat caaggccaac ttcaagatcc gccacaacat cgaggacggc
     1681 ggcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     1741 ctgcccgaca accactacct gagctaccag tccaagctga gcaaagaccc caacgagaag
     1801 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     1861 gagctgtaca agtaagtcga cggcgcgccg cggccgcgaa ttcgatatca taatcaacct
     1921 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     1981 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     2041 attttctcct ccttgtataa atcctggtta gttcttgcca cggcggaact catcgccgcc
     2101 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggctcga
     2161 gagatcttcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     2221 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     2281 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     2341 gggggaggat tgggaagaca atagcaggca tgagatctca cgtgcggacc gagcggccgc
     2401 aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg
     2461 ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc
     2521 gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg
     2581 gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag
     2641 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc
     2701 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     2761 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     2821 aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     2881 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     2941 actcaactct atctcgggct attcttttga tttataaggg attttgccga tttcggtcta
     3001 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     3061 gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     3121 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     3181 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     3241 atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     3301 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     3361 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     3421 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     3481 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     3541 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     3601 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     3661 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     3721 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     3781 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     3841 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     3901 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     3961 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     4021 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     4081 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     4141 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     4201 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     4261 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     4321 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     4381 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     4441 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     4501 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     4561 ttt
//
