LOCUS       sequence-423425-        5246 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of human DENND3 fragment V894 to E1198. 
            C-terminal HiBiT tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5246)
  AUTHORS   Ackloo S, Li F, Szewczyk M, Seitova A, Loppnau P, Zeng H, Xu J, 
            Ahmad S, Arnautova YA, Baghaie AJ, Beldar S, Bolotokova A, Centrella
            PA, Chau I, Clark MA, Cuozzo JW, Dehghani-Tafti S, Disch JS, Dong A,
            Dumas A, Feng JA, Ghiabi P, Gibson E, Gilmer J, Goldman B, Green SR,
            Guie MA, Guilinger JP, Harms N, Herasymenko O, Houliston S, 
            Hutchinson A, Kearnes S, Keefe AD, Kimani SW, Kramer T, Kutera M, 
            Kwak HA, Lento C, Li Y, Liu J, Loup J, Machado RAC, Mulhern CJ, 
            Perveen S, Righetto GL, Riley P, Shrestha S, Sigel EA, Silva M, 
            Sintchak MD, Slakman BL, Taylor RD, Thompson J, Torng W, 
            Underkoffler C, von Rechenberg M, Walsh RT, Watson I, Wilson DJ, 
            Wolf E, Yadav M, Yazdi AK, Zhang J, Zhang Y, Santhakumar V, Edwards 
            AM, Barsyte-Lovejoy D, Schapira M, Brown PJ, Halabelian L, 
            Arrowsmith CH
  TITLE     A Target Class Ligandability Evaluation of WD40 Repeat-Containing 
            Proteins.
  JOURNAL   J Med Chem. 2025 Jan 23;68(2):1092-1112. doi: 
            10.1021/acs.jmedchem.4c02010. Epub 2024 Nov 4.
  PUBMED    39495097
REFERENCE   2  (bases 1 to 5246)
  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"; doi: 
            "10.1021/acs.jmedchem.4c02010"; journalName: "J Med Chem"; date: 
            "2025-01-23- 23"; volume: "68"; issue: "2"; pages: "1092-1112"
COMMENT     Sequence Label: pBiT3.1-C_DENND3:894-1198
FEATURES             Location/Qualifiers
     source          1..5246
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    24..105
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     enhancer        338..641
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        642..845
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     796..816
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          981..1113
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1096..1115
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1157..1176
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1157..1175
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    2309..2430
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2320..2339)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(2545..2578)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2588..2635
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2667..3461
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2721..2740)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3331..3350
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(3619..4207)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3699..3718)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4333..4381
                     /note="synthetic polyadenylation signal"
     CDS             complement(4405..4803)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      4800..4809
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4834..5191)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4848..4983
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5000..5191
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5148..5168
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 aagacgcgtt aattaagacg tcggcagtga aaaaaatgct ttatttgtga aatttgtgat
       61 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
      121 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac
      181 ctctacaaat gtggtatggc tgattatgat ccgcggccgc aatacgtcga cgttatcagc
      241 tgacttcgta cgagagccta ggattatggc gcgccactag ttattaatag taatcaatta
      301 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      361 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      421 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      481 ctgcccactt ggcagtacat caagtgtatc atatgccaag tccgccccct attgacgtca
      541 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttacgg gactttccta
      601 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      661 acaccaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      721 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaata
      781 accccgcccc gttgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      841 gagctggttt agtgaaccgt cagatcacta gaatctttat tgcggtagtt tatcacagtt
      901 aaattgctaa cgcagtcagt gcttctgaca caacagtctc gaacttaagc tgcagaagtt
      961 ggtcgtgagg cactgggcag gtaagtatca aggttacaag acaggtttaa ggagaccaat
     1021 agaaactggg cttgtcgaga cagagaagac tcttgcgttt ctgataggca cctattggtc
     1081 ttactgacat ccactttgcc tttctctcca caggtgtcca ctcccagttc aattacagct
     1141 cttaaggcta gagtattaat acgactcact ataggggcga tcgcaacaag cttcgggaag
     1201 atctggagct agcggcgggg gatccggtgg gaattcgaca ccatggtgga cgtgctgctc
     1261 tacactccag ggcatcttga cccagccgaa aaagttgaag atgctcaccc caagttatgg
     1321 tgtgctctga gcgaaggcaa ggtgaccgtg ttcaatgctt cttcatggac catccaccag
     1381 cactccttta aagtgggcac tgcaaaagtg aactgcatgg tgatggccga ccagaaccag
     1441 gtgtgggttg gctcggaaga ctccgtcatc tacatcatca acgtccacag catgtcctgc
     1501 aacaagcagc tcacagccca ctgctccagt gtcacggatt tgattgtgca ggacggacag
     1561 gaggcaccca gcaacgtgta ctcgtgcagc atggacggca tggtgctggt gtggaatgtg
     1621 agcacactgc aggtgaccag ccgcttccag ctgccgcgag gtggcctgac gtccatcaga
     1681 ctgcacggcg gccgcctgtg gtgctgcaca ggtaacagca tcatggtcat gaaaatgaat
     1741 ggatccctcc atcaagaatt gaagattgag gagaacttca aagacaccag tacctccttc
     1801 ctggccttcc agctccttcc tgaggaggag cagctgtggg cggcctgtgc aggacgcagc
     1861 gaggtttaca tctggagcct gaaggacctg gcccagcccc cgcagagggt gcccctcgag
     1921 gactgctctg agatcaactg catgatccgg gtgaagaagc aggtctgggt gggcagccga
     1981 gggctggggc agggaacacc caaggggaaa atctacgtga ttgacgccga gaggaagacc
     2041 gtggagaagg agctggtggc gcacatggac accgtgagga cgctgtgctc ggctgaggac
     2101 agatacgtgc tgagtgggtc gggcagggag gaggggaaag tcgccatttg gaaaggcgaa
     2161 ggctcgagcg gtgggagctc cggtgtgagc ggctggcggc tgttcaagaa gattagctaa
     2221 gtttaaacgg ccgcgactct agactgcagg catgcaagct gatttggctg ctgccaccgc
     2281 tgagcaaggc cgcttcgagc agacatgata agatacattg atgagtttgg acaaaccaca
     2341 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt
     2401 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt
     2461 caggttcagg gggagatgtg ggaggttttt ttaagcaagt aaaacctcta caaatgtggt
     2521 aaaatcttaa ttaagacttc taatataact tcgtatagca tacattatac gaagttatgg
     2581 ttccactgtt gacaattaat catcggcata gtatatcggc atagtataat acgacaaggt
     2641 gaggaactaa acccaggagg cagatcatga ttgaacaaga tggattgcac gcaggttctc
     2701 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     2761 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     2821 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca
     2881 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     2941 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     3001 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     3061 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     3121 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     3181 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     3241 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     3301 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     3361 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     3421 agcgcatcgc cttctatcgc cttcttgacg agttcttctg attcgaaatg accgaccaag
     3481 cgacgcccaa ccggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     3541 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     3601 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     3661 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     3721 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     3781 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     3841 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     3901 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     3961 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     4021 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     4081 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     4141 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     4201 atttcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     4261 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttata
     4321 gtccggaatc gtcacacaaa aaaccaacac acagatgtaa tgaaaataaa gatattttat
     4381 tgcggccatc gtgatggcta gagtttagcc ctcccacaca taaccagagg gcagcaattc
     4441 acgaatccca actgccgtcg gctgtccatc actgtccttc actatggctt tgatcccagg
     4501 atgcagatcg agaagcacct gtcggcaccg tccgcagggg ctcaagatgc ccctgttctc
     4561 atttccgatc gcgacgatac aagtcaggtt gccagctgcc gcagcagcag cagtgcccag
     4621 caccacgagt tctgcacaag gtcccccagt aaaatgatat acattgacac cagtgaagat
     4681 gcggccgtcg ctagagagag ctgcgctggc gacgctgtag tcttcagaga tggggatgct
     4741 gttgattgta gccgttgctc tttcaatgag ggtggattct tcttgagaca aaggcttggc
     4801 catggtggcg ctagtgtcag aagaatcgag ctttttgcaa aagcctaggc ctccaaaaaa
     4861 gcctcctcac tacttctgga atagctcaga ggccgaggcg gcctcggcct ctgcataaat
     4921 aaaaaaaatt agtcagccat ggggcggaga atgggcggaa ctgggcggag ttaggggcgg
     4981 gatgggcgga gttaggggcg ggactatggt tgctgactaa ttgagatgca tgctttgcat
     5041 acttctgcct gctggggagc ctggggactt tccacacctg gttgctgact aattgagatg
     5101 catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc ctaactgaca
     5161 cacattccac agctggttct ttccgcctca gaaggtacct aaccaagttc ctctttcaga
     5221 ggttatttca ggccatggtg ctgcgc
//
