LOCUS       sequence-453373-        4740 bp    DNA     circular SYN 15-APR-2025
DEFINITION  AiE0780m is an enhancer sequence, designed to drive AAV-mediated 
            transgene expression in striatal direct pathway MSNs (D1-MSNs).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4740)
  AUTHORS   Hunker AC, Wirthlin ME, Gill G, Johansen NJ, Hooper M, Omstead V, 
            Taskin N, Weed N, Vargas S, Bendrick JL, Gore B, Ben-Simon Y, Bishaw
            Y, Opitz-Araya X, Martinez RA, Way S, Thyagarajan B, Lerma MN, Laird
            W, Sven O, Sanchez REA, Alexander JR, Amaya A, Amster A, Ayala A, 
            Baker PM, Barcelli T, Barta S, Bertagnolli D, Bielstein C, 
            Bishwakarma P, Bowlus J, Boyer G, Brouner K, Casian B, Casper T, 
            Chakka AB, Chakrabarty R, Clark M, Colbert K, Daniel S, Dawe T, 
            Departee M, DiValentin P, Donadio NP, Dotson NI, Dwivedi D, Egdorf 
            T, Fliss T, Gary A, Goldy J, Grasso C, Groce EL, Gudsnuk K, Han W, 
            Haradon Z, Hastings S, Helback O, Ho WV, Huang C, Johnson T, Jones 
            DL, Juneau Z, Kenney J, Leibly M, Li S, Liang E, Loeffler H, Lusk 
            NA, Madigan Z, Malloy J, Malone J, McCue R, Melchor J, Mich JK, 
            Moosman S, Morin E, Naidoo R, Newman D, Ngo K, Nguyen K, Oster AL, 
            Ouellette B, Oyama AA, Pena N, Pham T, Phillips E, Pom C, Potekhina 
            L, Ransford S, Reding M, Rette DF, Reynoldson C, Rimorin C, Rios 
            Sigler A, Rocha DB, Ronellenfitch K, Ruiz A, Sawyer L, Sevigny J, 
            Shapovalova NV, Shepard N, Shulga L, Soliman S, Staats B, Taormina 
            MJ, Tieu M, Wang Y, Wilkes J, Wood T, Zhou T, Williford A, Dee N, 
            Mollenkopf T, Ng L, Esposito L, Kalmbach B, Yao S, Ariza J, Mufti S,
            Smith K, Waters J, Ersing I, Patrick M, Zeng H, Lein ES, Kojima Y, 
            Horwitz G, Owen SF, Levi BP, Daigle TL, Tasic B, Bakken TE, Ting JT
  TITLE     Enhancer AAV toolbox for accessing and perturbing striatal cell 
            types and circuits.
  JOURNAL   bioRxiv [Preprint]. 2024 Sep 29:2024.09.27.615553. doi: 
            10.1101/2024.09.27.615553.
  PUBMED    39386678
REFERENCE   2  (bases 1 to 4740)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            [Preprint]. 2024 Sep 29:2024.09.27.615553. doi: 
            10.1101/2024.09.27.615553."
COMMENT     Sequence Label: AiP15746: pAAV-AiE0780m-minBG-mScarlet-WPRE3-B...
FEATURES             Location/Qualifiers
     source          1..4740
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     RBS             375..383
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     regulatory      773..782
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             779..1474
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(912..930)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1180..1199
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(1567..1587)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1774..1885
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2003..2143
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2003..2132
                     /label=AAV2 ITR
     rep_origin      2218..2673
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2305..2324)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2515..2536
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2690..2709)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2809..2831
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2869..2887)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2955..3059
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3060..3128,3129..3920)
                     /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(3278..3297)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4091..4679
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4580..4599
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgttgttt cacaaactac aggttcactg
      181 gagcacagcc atgccaacat ggcatctaag gctggtatga cctcgagacc cctccagcac
      241 tccttcgttc agccatagaa caaggagaca cagataaagt ctggagccgg gcccaggtac
      301 ctgaggctca gagatgagcc cgcagcgaag ctgacaactg actcattcct ccgacactct
      361 cccaccctcc cagctaagga ggtgtctgat gtgaacatgt tctgcctctt tgttgcctgc
      421 tgttgggggc gagggctttg ggtggatttt gaagagcaat tatgcttatt actggttagg
      481 atttctctgt ttgctgagtg cattaacttt tagcgcttcc catcactcat tcttcttcac
      541 accctcttgt cagttacaaa tgtgcaggtg gagagccacg aaaaaaccct gcagcaacgg
      601 caacagaaat cacagcaaca acaacaaatg aacagaaagg atagaaagca gatactcaaa
      661 gatggagaaa aagatactcc cgagaggctg agctcgggct gggcataaaa gtcagggcag
      721 agccatctat tgcttacatt tgcttctggg atccagatct ttcgaagcta gcgccaccat
      781 ggtgagcaag ggcgaggcag tgatcaagga gttcatgcgg ttcaaggtgc acatggaggg
      841 ctccatgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg
      901 cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttct cctgggacat
      961 cctgtcccct cagttcatgt acggctccag ggccttcacc aagcaccccg ccgacatccc
     1021 cgactactat aagcagtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga
     1081 ggacggcggc gccgtgaccg tgacccagga cacctccctg gaggacggca ccctgatcta
     1141 caaggtgaag ctccgcggca ccaacttccc tcctgacggc cccgtaatgc agaagaagac
     1201 aatgggctgg gaagcgtcca ccgagcggtt gtaccccgag gacggcgtgc tgaagggcga
     1261 cattaagatg gccctgcgcc tgaaggacgg cggccgctac ctggcggact tcaagaccac
     1321 ctacaaggcc aagaagcccg tgcagatgcc cggcgcctac aacgtcgacc gcaagttgga
     1381 catcacctcc cacaacgagg actacaccgt ggtggaacag tacgaacgct ccgagggccg
     1441 ccactccacc ggcggcatgg acgagctgta caagtaagtc gacggcgcgc cgcggccgcg
     1501 aattcgatat cataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     1561 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     1621 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tagttcttgc
     1681 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     1741 cactgacaat tccgtggctc gagagatctt cgactgtgcc ttctagttgc cagccatctg
     1801 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
     1861 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
     1921 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgagatct
     1981 cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg
     2041 cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc
     2101 cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat
     2161 tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg
     2221 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     2281 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     2341 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     2401 ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat
     2461 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     2521 tcttgttcca aactggaaca acactcaacc ctatctcggg ctattctttt gatttataag
     2581 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     2641 cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct
     2701 ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac
     2761 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     2821 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac
     2881 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     2941 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     3001 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     3061 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     3121 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     3181 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     3241 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     3301 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     3361 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     3421 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     3481 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     3541 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     3601 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     3661 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     3721 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     3781 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     3841 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     3901 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     3961 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     4021 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     4081 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     4141 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     4201 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     4261 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     4321 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     4381 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     4441 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     4501 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     4561 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     4621 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     4681 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
//
