LOCUS       sequence-460618-        5800 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  DLX2.0 is an enhancer sequence, designed to drive cytosolic SYFP2 
            and nuclear mScarlet expression in forebrain GABAergic neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5800)
  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 5800)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 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: AiP13798 -
            pAAV-DLX2.0-minBG-SYFP2-P2A-mScarle...
FEATURES             Location/Qualifiers
     source          1..5800
                     /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      636..645
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(642..644,645..647,648..1358)
                     /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(687..708)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(948..967)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1295..1316
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1368..1424
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             1440..2135
                     /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(1573..1591)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1841..1860
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             2166..2543
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(2627..2647)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2834..2945
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3063..3203
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3063..3192
                     /label=AAV2 ITR
     rep_origin      3278..3733
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3365..3384)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3575..3596
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3750..3769)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3869..3891
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3929..3947)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4015..4119
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4120..4188,4189..4980)
                     /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(4338..4357)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5151..5739
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5640..5659
                     /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 ctagcgctac cggtcgccac catggtgagc aagggcgagg
      661 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
      721 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagc
      781 tgatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgggct
      841 acggcgtgca gtgcttcgcc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
      901 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
      961 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     1021 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
     1081 acagccacaa cgtctatatc accgccgaca agcagaagaa cggcatcaag gccaacttca
     1141 agatccgcca caacatcgag gacggcggcg tgcagctcgc cgaccactac cagcagaaca
     1201 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc taccagtcca
     1261 agctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
     1321 ccgccgggat cactctcggc atggacgagc tgtacaaggg aagcggagcc actaacttct
     1381 ccctgttgaa acaagcaggg gatgtcgaag agaatcccgg gccagcaccg ggctccacca
     1441 tggtgagcaa gggcgaggca gtgatcaagg agttcatgcg gttcaaggtg cacatggagg
     1501 gctccatgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg
     1561 gcacccagac cgccaagctg aaggtgacca agggtggccc cctgcccttc tcctgggaca
     1621 tcctgtcccc tcagttcatg tacggctcca gggccttcac caagcacccc gccgacatcc
     1681 ccgactacta taagcagtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg
     1741 aggacggcgg cgccgtgacc gtgacccagg acacctccct ggaggacggc accctgatct
     1801 acaaggtgaa gctccgcggc accaacttcc ctcctgacgg ccccgtaatg cagaagaaga
     1861 caatgggctg ggaagcgtcc accgagcggt tgtaccccga ggacggcgtg ctgaagggcg
     1921 acattaagat ggccctgcgc ctgaaggacg gcggccgcta cctggcggac ttcaagacca
     1981 cctacaaggc caagaagccc gtgcagatgc ccggcgccta caacgtcgac cgcaagttgg
     2041 acatcacctc ccacaacgag gactacaccg tggtggaaca gtacgaacgc tccgagggcc
     2101 gccactccac cggcggcatg gacgagctgt ataagtccgg actcagatct ggaggctccg
     2161 gaggcccaga gccagcgaag tctgctcccg ccccgaaaaa gggctccaag aaggcggtga
     2221 ctaaggcgca gaagaaaggc ggcaagaagc gcaagcgcag ccgcaaggag agctattcca
     2281 tctatgtgta taaggttctg aagcaggtcc accctgacac cggcatttcg tccaaggcca
     2341 tgggcatcat gaactcgttt gtgaacgaca ttttcgagcg catcgcaggt gaggcttccc
     2401 gcctggcgca ttacaacaag cgctcgacca tcacctccag ggagatccag acggccgtgc
     2461 gcctgctgct gcctggggag ttggccaagc acgccgtgtc cgagggtact aaggccatca
     2521 ccaagtacac cagcgctaag taaggcgcgc ccctgcaggg aattcgatat cataatcaac
     2581 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
     2641 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
     2701 tcattttctc ctccttgtat aaatcctggt tagttcttgc cacggcggaa ctcatcgccg
     2761 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggctc
     2821 gagagatctt cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     2881 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     2941 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     3001 aagggggagg attgggaaga caatagcagg catgagatct cacgtgcgga ccgagcggcc
     3061 gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga
     3121 ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga
     3181 gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta cgcatctgtg
     3241 cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta
     3301 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccttagcg
     3361 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     3421 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     3481 aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     3541 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     3601 acactcaact ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggtc
     3661 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     3721 acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     3781 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     3841 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     3901 tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     3961 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     4021 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     4081 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     4141 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     4201 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     4261 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     4321 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     4381 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     4441 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     4501 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     4561 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     4621 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     4681 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     4741 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     4801 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     4861 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     4921 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     4981 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     5041 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     5101 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     5161 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     5221 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     5281 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     5341 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     5401 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     5461 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     5521 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     5581 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     5641 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     5701 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     5761 ttacggttcc tggccttttg ctggcctttt gctcacatgt
//
