LOCUS       sequence-466241-        4701 bp DNA     circular SYN 24-MAR-2025
DEFINITION  AiE2126m 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 4701)
  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 4701)
  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..4701
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(561..1421)
                     /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     1184..1203
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1422..1526)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1594..1612
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1650..1672)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1772..1791
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(1808..2263)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1945..1966)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2157..2176
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2338..2478
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2349..2478
                     /label=AAV2 ITR
     polyA_signal    2596..2707
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2894..2914
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3007..3723)
                     /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(3049..3070)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3398..3417
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3657..3678
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3720..3729
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   4312..4441
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(join(4503..4701,1..390))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4583..4602)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
       61 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
      121 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
      181 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
      241 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
      301 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
      361 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
      421 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
      481 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
      541 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
      601 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
      661 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
      721 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
      781 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
      841 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
      901 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
      961 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     1021 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     1081 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     1141 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     1201 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     1261 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     1321 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     1381 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     1441 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     1501 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     1561 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     1621 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
     1681 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
     1741 gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg
     1801 caccataaaa ttgtaaacgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
     1861 agctcatttt ttaaccaata gaccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     1921 cccgagatag agttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     1981 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
     2041 tcacccaaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
     2101 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
     2161 aagaaagcga aaggagcggg cgctaaggcg ctggcaagtg tagcggtcac gctgcgcgta
     2221 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtactatgg ttgctttgac
     2281 gtatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgcccct
     2341 gcaggcagct gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg
     2401 cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga gtggccaact
     2461 ccatcactag gggttcctgc ggccgctcgg tccgcacgtg agatctcatg cctgctattg
     2521 tcttcccaat cctccccctt gctgtcctgc cccaccccac cccccagaat agaatgacac
     2581 ctactcagac aatgcgatgc aatttcctca ttttattagg aaaggacagt gggagtggca
     2641 ccttccaggg tcaaggaagg cacgggggag gggcaaacaa cagatggctg gcaactagaa
     2701 ggcacagtcg aagatctctc gagccacgga attgtcagtg cccaacagcc gagcccctgt
     2761 ccagcagcgg gcaaggcagg cggcgatgag ttccgccgtg gcaagaacta accaggattt
     2821 atacaaggag gagaaaatga aagccatacg ggaagcaata gcatgataca aaggcattaa
     2881 agcagcgtat ccacatagcg taaaaggagc aacatagtta agaataccag tcaatctttc
     2941 acaaattttg taatccagag gttgattatg atatcgaatt cgcggccgcg gcgcgccgtc
     3001 gacttacttg tacagctcgt ccatgccgag agtgatcccg gcggcggtca cgaactccag
     3061 caggaccatg tgatcgcgct tctcgttggg gtctttgctc agcttggact ggtagctcag
     3121 gtagtggttg tcgggcagca gcacggggcc gtcgccgatg ggggtgttct gctggtagtg
     3181 gtcggcgagc tgcacgccgc cgtcctcgat gttgtggcgg atcttgaagt tggccttgat
     3241 gccgttcttc tgcttgtcgg cggtgatata gacgttgtgg ctgttgtagt tgtactccag
     3301 cttgtgcccc aggatgttgc cgtcctcctt gaagtcgatg cccttcagct cgatgcggtt
     3361 caccagggtg tcgccctcga acttcacctc ggcgcgggtc ttgtagttgc cgtcgtcctt
     3421 gaagaagatg gtgcgctcct ggacgtagcc ttcgggcatg gcggacttga agaagtcgtg
     3481 ctgcttcatg tggtcggggt agcgggcgaa gcactgcacg ccgtagccca gggtggtcac
     3541 gagggtgggc cagggcacgg gcagcttgcc ggtggtgcag atcagcttca gggtcagctt
     3601 gccgtaggtg gcatcgccct cgccctcgcc ggacacgctg aacttgtggc cgtttacgtc
     3661 gccgtccagc tcgaccagga tgggcaccac cccggtgaac agctcctcgc ccttgctcac
     3721 catggtggcg accggtagcg ctagcttcga aagatctgga tcccagaagc aaatgtaagc
     3781 aatagatggc tctgccctga cttttatgcc cagcccgagc tcctaacaag acagacacag
     3841 acaggtacgc actgcttgat gctctctcta cagatgcaga atctgatgta ggactgtgag
     3901 acactgtagc tctccccact gacttcatct tgccataatt gtgttctcta acaattgtta
     3961 actctagaga accagacatc ctgtgagtct tggcattgtc agagaagtct catgcttcct
     4021 gtatgactca gcctagtcct tgcgtctggt cagtctctag atgttttcct gtctgtagtt
     4081 gtgaagctgg cttgtgacgt ctgctctgtg ggccaggccg aggcttcagg aaagaactag
     4141 cagtgctcca ggaaggaaaa ggtggagcca caggctgctc tcctcatgca ccactagcct
     4201 cctctgtcag ttgctcttca gaggccctgg acaacaaagt tctggtttct atctgtgaac
     4261 agaagctgca gcacttcagg tgagagcaga gaaaagcacg cgtgcggccg caggaacccc
     4321 tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac
     4381 caaaggtcgc ccgacgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag
     4441 gacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     4501 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     4561 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     4621 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     4681 agcgtggcgc tttctcatag c
//
