LOCUS       sequence-470927-        4763 bp    DNA     circular SYN 30-APR-2025
DEFINITION  AiE0017m_3xC1 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 4763)
  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 4763)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 30, 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: AiP13551 -
            pAAV-AiE0017m_3xC1-minBG-SYFP2-WPRE...
FEATURES             Location/Qualifiers
     source          1..4763
                     /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      775..784
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(781..783,784..786,787..1497)
                     /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(826..847)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1087..1106)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1434..1455
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1590..1610)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1797..1908
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2026..2166
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2026..2155
                     /label=AAV2 ITR
     rep_origin      2241..2696
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2328..2347)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2538..2559
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2713..2732)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2832..2854
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2892..2910)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2978..3082
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3083..3151,3152..3943)
                     /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(3301..3320)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4114..4702
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4603..4622
                     /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 gcgttgtgtg tggtcttagg gacagggggt ggctggcagg
      181 caggcagtgg cgggaggcac agctgtgcag gaggacggac tgctggcatc agcaagcgcc
      241 cctcctcccc cccacactaa acaatcacgg cacaaatatg caaatggtat aattactgtt
      301 atttgttttg ctgatataag tgttgtgtgt ggtcttaggg acagggggtg gctggcaggc
      361 aggcagtggc gggaggcaca gctgtgcagg aggacggact gctggcatca gcaagcgccc
      421 ctcctccccc ccacactaaa caatcacggc acaaatatgc aaatggtata attactgtta
      481 tttgttttgc tgatataagt gttgtgtgtg gtcttaggga cagggggtgg ctggcaggca
      541 ggcagtggcg ggaggcacag ctgtgcagga ggacggactg ctggcatcag caagcgcccc
      601 tcctcccccc cacactaaac aatcacggca caaatatgca aatggtataa ttactgttat
      661 ttgttttgct gatataagtg tgagctcggg ctgggcataa aagtcagggc agagccatct
      721 attgcttaca tttgcttctg ggatccagat ctttcgaagc tagcgctacc ggtcgccacc
      781 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
      841 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
      901 ggcaagctga ccctgaagct gatctgcacc accggcaagc tgcccgtgcc ctggcccacc
      961 ctcgtgacca ccctgggcta cggcgtgcag tgcttcgccc gctaccccga ccacatgaag
     1021 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
     1081 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     1141 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     1201 aagctggagt acaactacaa cagccacaac gtctatatca ccgccgacaa gcagaagaac
     1261 ggcatcaagg ccaacttcaa gatccgccac aacatcgagg acggcggcgt gcagctcgcc
     1321 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     1381 tacctgagct accagtccaa gctgagcaaa gaccccaacg agaagcgcga tcacatggtc
     1441 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa
     1501 gtcgacggcg cgccgcggcc gcgaattcga tatcataatc aacctctgga ttacaaaatt
     1561 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     1621 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     1681 tataaatcct ggttagttct tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     1741 tggacagggg ctcggctgtt gggcactgac aattccgtgg ctcgagagat cttcgactgt
     1801 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     1861 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     1921 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     1981 agacaatagc aggcatgaga tctcacgtgc ggaccgagcg gccgcaggaa cccctagtga
     2041 tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg
     2101 tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc
     2161 tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     2221 tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     2281 ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt
     2341 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     2401 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg
     2461 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     2521 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca actctatctc
     2581 gggctattct tttgatttat aagggatttt gccgatttcg gtctattggt taaaaaatga
     2641 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg
     2701 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
     2761 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     2821 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     2881 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     2941 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     3001 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     3061 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     3121 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     3181 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     3241 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     3301 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     3361 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     3421 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     3481 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     3541 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     3601 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     3661 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     3721 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     3781 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     3841 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     3901 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     3961 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     4021 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     4081 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     4141 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     4201 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     4261 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     4321 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     4381 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     4441 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     4501 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     4561 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     4621 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     4681 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     4741 ttgctggcct tttgctcaca tgt
//
