LOCUS       sequence-466222-        6225 bp DNA     circular SYN 24-MAR-2025
DEFINITION  AiE2069m 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 6225)
  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 6225)
  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..6225
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(91..951)
                     /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     714..733
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(952..1056)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1124..1142
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1180..1202)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1302..1321
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(1338..1793)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1475..1496)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1687..1706
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   1868..2008
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1879..2008
                     /label=AAV2 ITR
     polyA_signal    2048..2524
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     2306..2325
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     misc_feature    2556..3144
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             2662..2673
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     3071..3091
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3169..4467)
                     /codon_start=1
                     /product="nuclear-targeted site-specific recombinase"
                     /label=FLPo
                     /note="FLPo is a mouse codon-optimized fusion between the 
                     SV40 nuclear localization signal and a thermostable version
                     of the Saccharomyces cerevisiae site-specific recombinase 
                     FLP.  Recombination occurs at FRT sequences."
                     /translation="MAPKKKRKVMSQFDILCKTPPKVLVRQFVERFERPSGEKIASCAA
                     ELTYLCWMITHNGTAIKRATFMSYNTIISNSLSFDIVNKSLQFKYKTQKATILEASLKK
                     LIPAWEFTIIPYNGQKHQSDITDIVSSLQLQFESSEEADKGNSHSKKMLKALLSEGESI
                     WEITEKILNSFEYTSRFTKTKTLYQFLFLATFINCGRFSDIKNVDPKSFKLVQNKYLGV
                     IIQCLVTETKTSVSRHIYFFSARGRIDPLVYLDEFLRNSEPVLKRVNRTGNSSSNKQEY
                     QLLKDNLVRSYNKALKKNAPYPIFAIKNGPKSHIGRHLMTSFLSMKGLTELTNVVGNWS
                     DKRASAVARTTYTHQITAIPDHYFALVSRYYAYDPISKEMIALKDETNPIEEWQHIEQL
                     KGSAEGSIRYPAWNGIISQEVLDYLSSYINRRI"
     regulatory      4464..4473
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   5366..5495
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(5557..6145)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5637..5656)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
       61 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
      121 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
      181 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
      241 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
      301 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
      361 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
      421 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
      481 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
      541 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
      601 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
      661 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
      721 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
      781 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
      841 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
      901 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
      961 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     1021 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     1081 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     1141 gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
     1201 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
     1261 gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt
     1321 actgagagtg caccataaaa ttgtaaacgt taatattttg ttaaaattcg cgttaaattt
     1381 ttgttaaatc agctcatttt ttaaccaata gaccgaaatc ggcaaaatcc cttataaatc
     1441 aaaagaatag cccgagatag agttgagtgt tgttccagtt tggaacaaga gtccactatt
     1501 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
     1561 acgtgaacca tcacccaaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
     1621 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
     1681 aaaggaaggg aagaaagcga aaggagcggg cgctaaggcg ctggcaagtg tagcggtcac
     1741 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtactatgg
     1801 ttgctttgac gtatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
     1861 aggcgcccct gcaggcagct gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc
     1921 gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga
     1981 gtggccaact ccatcactag gggttcctgc ggccgctcgg tccgcacgtg gttacctaca
     2041 aaatcagaag gacagggaag ggagcagtgg ttcacgcctg taatcccagc aatttgggag
     2101 gccaaggtgg gtagatcacc tgagattagg agttggagac cagcctggcc aatatggtga
     2161 aaccccgtct ctaccaaaaa aacaaaaatt agctgagcct ggtcatgcat gcctggaatc
     2221 ccaacaactc gggaggctga ggcaggagaa tcgcttgaac ccaggaggcg gagattgcag
     2281 tgagccaaga ttgtgccact gcactccagc ttggttccca atagaccccg caggccctac
     2341 aggttgtctt cccaacttgc cccttgctcc ataccacccc cctccacccc ataatattat
     2401 agaaggacac ctagtcagac aaaatgatgc aacttaattt tattaggaca aggctggtgg
     2461 gcactggagt ggcaacttcc agggccagga gaggcactgg ggaggggtca cagggatgcc
     2521 acccgtagat ctctcgagca gcgctcggta tcgatgcggg gaggcggccc aaagggagat
     2581 ccgactcgtc tgagggcgaa ggcgaagacg cggaagaggc cgcagagccg gcagcaggcc
     2641 gcgggaagga aggtccgctg gattgagggc cgaagggacg tagcagaagg acgtcccgcg
     2701 cagaatccag gtggcaacat aggcgagcag ccaaggaaag gacgatgatt tccccgacaa
     2761 caccacggaa ttgtcagtgc ccaacagccg agcccctgtc cagcagcggg caaggcaggc
     2821 ggcgatgagt tccgccgtgg caatagggag ggggaaagcg aaagtcccgg aaaggagctg
     2881 acaggtggtg gcaatgcccc aaccagtggg ggttgcgtca gcaaacacag tgcacaccac
     2941 gccacgttgc ctgacaacgg gccacaactc ctcataaaga gacagcaacc aggatttata
     3001 caaggaggag aaaatgaaag ccatacggga agcaatagca tgatacaaag gcattaaagc
     3061 agcgtatcca catagcgtaa aaggagcaac atagttaaga ataccagtca atctttcaca
     3121 aattttgtaa tccagaggtt gattatcgat aagcttgata tcgaattctc agatccgcct
     3181 gttgatgtag ctgctcaggt agtccagcac ctcctggctg atgatgccgt tccaggcggg
     3241 gtatctgatg ctgccctcgg cgctgccctt cagctgctcg atgtgctgcc actcctcgat
     3301 ggggttggtc tcgtccttca gggcgatcat ctccttgctg atggggtcgt aggcgtagta
     3361 cctggacacc agggcgaagt agtggtcggg gatggcggtg atctggtggg tgtaggtggt
     3421 cctggccacg gcggaggccc tcttgtcgct ccagttgccc accacgtttg tcagctcggt
     3481 caggcccttc atgctcagaa agctggtcat caggtgcctg ccgatgtggc tcttagggcc
     3541 gttcttgata gcgaagatgg ggtagggggc gttcttcttc agggccttgt tgtagctgcg
     3601 caccaggttg tccttcagca gctggtactc ctgcttgttg ctgctgctgt tgccggtcct
     3661 gttcactctc ttcagcacgg gctcgctgtt cctcaggaac tcgtccaggt acaccagggg
     3721 gtcgatcctg cctctggcgc tgaaaaagta gatgtgcctg gacacgcttg tcttggtctc
     3781 ggtcaccagg cactgaatga tcacgcccag gtacttgttc tgcaccagct tgaagctctt
     3841 ggggtccacg ttcttgatgt cgctgaacct gccgcagttg atgaatgtgg ccaggaacag
     3901 gaactggtac agggtcttgg tcttggtgaa cctgctggtg tactcgaagc tgttcaggat
     3961 cttctcggtg atctcccaga tgctctcgcc ctcggacagc agggccttca gcatcttctt
     4021 gctgtggctg ttgcccttgt cggcctcctc gctgctctcg aactgcagct gcaggctgga
     4081 cacgatgtcg gtgatgtcgc tctggtgctt ctggccgttg taagggatga tggtgaactc
     4141 ccaggcgggg atcagcttct tcaggctggc ctccaggatg gtggccttct gggtcttgta
     4201 cttgaactgc aggctcttgt tcacgatgtc gaagctcagg ctgttgctga tgatggtgtt
     4261 gtagctcatg aaggtggccc tcttgatggc ggtgccgttg tgggtgatca tccagcacag
     4321 gtaggtcagc tcggcggcac agctggcgat cttctcgccg ctgggcctct cgaatctctc
     4381 cacgaactgc cgcaccagca ccttgggggg ggtcttgcac aggatgtcga actggctcat
     4441 caccttcctc ttcttcttag gagccatggt ggccacgcca gaagcaaatg taagcaatag
     4501 atggctctgc cctgactttt atgcccagcc cgagctctac atcttacact ccatgctccc
     4561 aggcaagcct gggtgttctc aaagtagata gtgcccatcc ccaatagatg acccgttttt
     4621 aaaggggatg catctggaat acttaacaga gtggaacaga gtaatgcctc aaagggtcac
     4681 ctgtgctgaa ggattttaaa ccctggtcct ttttctaagt agctccactg tagattactt
     4741 accaggatct tcttctccag tggagactgc tggccttctg aaatcctagg gtctctggat
     4801 tcttttgccc aagccaatct aaaagataaa gcaaaggtag attggagtag gggagacacc
     4861 acatgtttgc tcacttatca ttccagtcag agatctcaat gatttggcca agaacatggg
     4921 ccatttctcc atcggggaaa catttgcatt tcccatggca caaaaaacaa cgagcatttc
     4981 ttatatatga actaaaagca tacctctcct gtggactcaa ggcaaaccac tcagaatcat
     5041 ttatttggca tggacagaag acagaaatag gctctttggt gagggatggc ctaaggacac
     5101 ccagaccaga ttgttcagtt tcaaagcctt ttttgtcttc tgagataatt ctagtctctc
     5161 gtttgtccag ctcattgtct ttctgcactt tgtccagcca ggccctctgt gcttctatca
     5221 gcacatacag catctttgcc ttgctttgta ctgaaattct attcctcttt gatcatctgg
     5281 ccctcggctt gactgactga tactgttaat catccttcac cctgaatcca ttttgagact
     5341 attccctgct gacgcgtgcg gccgcaggaa cccctagtga tggagttggc cactccctct
     5401 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcggc
     5461 ctcagtgagc gagcgagcgc gcagctgcct gcaggacatg tgagcaaaag gccagcaaaa
     5521 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     5581 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     5641 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     5701 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     5761 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     5821 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     5881 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     5941 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     6001 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     6061 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     6121 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     6181 tcagtggaac gaaaactcac gttaagggat tttggtcatg agatt
//
