LOCUS       sequence-485889-        4593 bp    DNA     circular SYN 12-SEP-2025
DEFINITION  DLX2.0 is an enhancer sequence, designed to drive nuclear mScarlet 
            expression in forebrain GABAergic neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4593)
  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 (preprint).
  JOURNAL   bioRxiv [Preprint]. 2024 Sep 26:2024.06.10.597244. doi: 
            10.1101/2024.06.10.597244.
  PUBMED    38915722
REFERENCE   2  (bases 1 to 4593)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 12, 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: AiP14211 - pAAV-DLX2.0-minBG-mScarlet-WPRE3-BG...
FEATURES             Location/Qualifiers
     source          1..4593
                     /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      626..635
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             632..1327
                     /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(765..783)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1033..1052
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(1420..1440)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1627..1738
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1856..1996
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1856..1985
                     /label=AAV2 ITR
     rep_origin      2071..2526
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2158..2177)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2368..2389
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2543..2562)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2662..2684
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2722..2740)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2808..2912
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2913..2981,2982..3773)
                     /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(3131..3150)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3944..4532
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4433..4452
                     /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 gcgtggtacc ctaaataaag atggcttttt agtattaaaa
      181 gtggaagaaa attacaggta attatctttg acggtaaaaa cgctgtaatc agcgggctac
      241 atgaaaaatt actctaatta tggctgcatt taagagaatg gctaaataaa gatggctttt
      301 tagtattaaa agtggaagaa aattacaggt aattatcttt gacggtaaaa acgctgtaat
      361 cagcgggcta catgaaaaat tactctaatt atggctgcat ttaagagaat ggctaaataa
      421 agatggcttt ttagtattaa aagtggaaga aaattacagg taattatctt tgacggtaaa
      481 aacgctgtaa tcagcgggct acatgaaaaa ttactctaat tatggctgca tttaagagaa
      541 tggagctcgg gctgggcata aaagtcaggg cagagccatc tattgcttac atttgcttct
      601 gggatccaga tctttcgaag ctagcgccac catggtgagc aagggcgagg cagtgatcaa
      661 ggagttcatg cggttcaagg tgcacatgga gggctccatg aacggccacg agttcgagat
      721 cgagggcgag ggcgagggcc gcccctacga gggcacccag accgccaagc tgaaggtgac
      781 caagggtggc cccctgccct tctcctggga catcctgtcc cctcagttca tgtacggctc
      841 cagggccttc accaagcacc ccgccgacat ccccgactac tataagcagt ccttccccga
      901 gggcttcaag tgggagcgcg tgatgaactt cgaggacggc ggcgccgtga ccgtgaccca
      961 ggacacctcc ctggaggacg gcaccctgat ctacaaggtg aagctccgcg gcaccaactt
     1021 ccctcctgac ggccccgtaa tgcagaagaa gacaatgggc tgggaagcgt ccaccgagcg
     1081 gttgtacccc gaggacggcg tgctgaaggg cgacattaag atggccctgc gcctgaagga
     1141 cggcggccgc tacctggcgg acttcaagac cacctacaag gccaagaagc ccgtgcagat
     1201 gcccggcgcc tacaacgtcg accgcaagtt ggacatcacc tcccacaacg aggactacac
     1261 cgtggtggaa cagtacgaac gctccgaggg ccgccactcc accggcggca tggacgagct
     1321 gtacaagtaa gtcgacggcg cgccgcggcc gcgaattcga tatcataatc aacctctgga
     1381 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     1441 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     1501 ctcctccttg tataaatcct ggttagttct tgccacggcg gaactcatcg ccgcctgcct
     1561 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg ctcgagagat
     1621 cttcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
     1681 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
     1741 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
     1801 aggattggga agacaatagc aggcatgaga tctcacgtgc ggaccgagcg gccgcaggaa
     1861 cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg
     1921 cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg
     1981 cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     2041 tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg
     2101 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc
     2161 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     2221 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     2281 ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     2341 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     2401 actctatctc gggctattct tttgatttat aagggatttt gccgatttcg gtctattggt
     2461 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta
     2521 caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc
     2581 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     2641 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     2701 cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     2761 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     2821 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     2881 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     2941 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     3001 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     3061 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     3121 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     3181 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     3241 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     3301 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     3361 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     3421 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     3481 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     3541 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     3601 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     3661 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     3721 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     3781 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     3841 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     3901 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     3961 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     4021 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     4081 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     4141 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     4201 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     4261 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     4321 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     4381 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     4441 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     4501 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     4561 tcctggcctt ttgctggcct tttgctcaca tgt
//
