LOCUS       sequence-423408-        6828 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human DNAI4. N-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6828)
  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 6828)
  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-N_DNAI4:1-848
FEATURES             Location/Qualifiers
     source          1..6828
                     /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    3891..4012
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3902..3921)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(4127..4160)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        4170..4217
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4249..5043
                     /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(4303..4322)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4913..4932
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(5201..5789)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5281..5300)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5915..5963
                     /note="synthetic polyadenylation signal"
     CDS             complement(5987..6385)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      6382..6391
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(6416..6773)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6430..6565
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        6582..6773
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     6730..6750
                     /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 ataggggtta gcgatcgcca tggtgagcgg
     1201 ctggcggctg ttcaagaaga ttagcgggag ctccggtggc tcgagcggta tgacgcccgg
     1261 caaacattcc ggagcctcgg cccgagccgc taacgcagga gcttgggggt acagggactt
     1321 cagaggcggc caaaaaaagg ggtggtggac cactccccag ctggtcgcca ccatgccagt
     1381 ctctccggca ggcagtcaca agcagcagaa cttcgggttg aacaatgcca cacaaccaaa
     1441 gaagtctatt agcttttttg ctacaatgaa agcaacttca gtgaaaggat atactggtgc
     1501 aaatcaaagc agaatggctg tgtccaaaac cgtgcttatt ccacctgaac tgaaaactgt
     1561 agaaaaacca aatcccaata taaagacaac acaggtattt gacataaatg gaactgatgt
     1621 tactccccga cctctttacc atccagatcc acttactggt acagcaaaac caagtaaact
     1681 cttgacatca caagaaggat cacttggatc agaatttata tcttcctata gcctttatca
     1741 gaatacaata aatcctagta cgttagggca gtttacaagg tcagttttag gaagcagtac
     1801 agtttctaag tcaagtgtat cagcaagtga atcaatagca gaagacctgg aagaaccatc
     1861 ctataaacgg gaaagattga ctagtttcac agatttgcaa gttataaggg cagcacctga
     1921 aaaaattgta acaaaagaag acctggagaa gaatatagag ataatactca cagagacaga
     1981 aacactgaga ttttttgact tgcccacagt catggtctct gtagaatctg aagaagctga
     2041 gaaagtaact cagagaaaca agaattatga agtcctttgt agaaacagat taggcaatga
     2101 cctatatgtt gaaaggatga tgcagacttt caatggagca ccaaagaata aagatgttca
     2161 atgtgataaa atcataatgg aagataaagg cataatgtcc actgcctggg atttgtatga
     2221 ttcttacaat gctatggaac ttgtatcttt atcagtaaaa caatctgtgg ttgagtcaag
     2281 tagtaaagca aatgtacttc ctaaagatca ggaccaaaga ttgccaggga gcactacaga
     2341 aaaaaatagt gaaactagtt ctctaatgga catagaaaat gtaattctgg caaaaatcca
     2401 tgaagatgag gaagaccact cagatgcaat attaaaatct gacaaatttc atcaggactt
     2461 attttttatg gaacgggttc tgatggaaaa tatatttcag cccaaacttg cagcttatcg
     2521 tcagcttcct gttttaaaag aacctgaacc tgaagagcct gaagatgttt tagaaagtgc
     2581 aaaacatgaa gaggtagagg aagaatctaa gaaggaggag gaagaagaaa tacatgcaga
     2641 agaatcaaca atacccgcca acttggaacg actttggtct ttttcctgtg acttaaccaa
     2701 aggcctcaat gtgagcagcc ttgcctggaa taaaacaaat ccagatcttt tggctgttgg
     2761 ctatgggcac tttggattta aagagcaaaa aagaggactg gcttgctgct ggtcaataaa
     2821 gaatcccatg tggccagaac gtatttatca gagtccatat ggagttactg ctgtggattt
     2881 ttcaattgga gcacctaacc ttttagccgt tggctatcac aatggcacaa ttgcaattta
     2941 caatgtacgg agcaacagta atgttccagt tctggatagt agtgaatcac ctcaaaaaca
     3001 tttgggacct gtatggcaac tacagtggat agaacaagat cgaggaacaa caggagatgg
     3061 caaaagagaa atactagttt ctatatcagc agatggaaga atctccaaat gggttatacg
     3121 aaaaggacta gactgttatg atttgatgcg attaaagaga actacagctg ccagtaacaa
     3181 aaaaggaggg gaaaaggaaa agaaagatga agctttgata tctcgacagg ctcctggaat
     3241 gtgttttgct tttcatccca aggacacaaa tatctatttg gctggcactg aagaaggtca
     3301 tattcacaaa tgttcttgtt catataatga acaatactta gatacctaca gaggacataa
     3361 gggtccagtg tataaagtga catggaatcc attttgtcat gatgtatttt taagctgttc
     3421 tgcagattgg ggtgttatta tatggcaaca ggagaatgtc aagccatctt tgagttttta
     3481 tccagctact tctgttgttt acgacgttgc ctggtctcca aaatcatcct atatatttgc
     3541 agctgcaaat gagaacaggg tggagatttg ggaccttcat atcagcactt tggaccctct
     3601 gattgtgaat actgctaacc ctggaatcaa gttcacaacc attctctttg ccaaacaaac
     3661 agattgcctt ctggtaggag acagtgatgg acaggtttct gtgtatgaac tgagaaatat
     3721 gcctactgtt ttggaaactg gccggggaga tataatggat actttgcttg gatccaagtc
     3781 aaaccaatca gcataatcta gaggtcaagc ttcgggggtt taaacctgca ggcatgcaag
     3841 ctgatttggc tgctgccacc gctgagcaag gccgcttcga gcagacatga taagatacat
     3901 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     3961 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
     4021 caattgcatt cattttatgt ttcaggttca gggggagatg tgggaggttt ttttaagcaa
     4081 gtaaaacctc tacaaatgtg gtaaaatctt aattaagact tctaatataa cttcgtatag
     4141 catacattat acgaagttat ggttccactg ttgacaatta atcatcggca tagtatatcg
     4201 gcatagtata atacgacaag gtgaggaact aaacccagga ggcagatcat gattgaacaa
     4261 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     4321 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     4381 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     4441 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     4501 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     4561 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     4621 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     4681 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     4741 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     4801 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     4861 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     4921 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     4981 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     5041 tgattcgaaa tgaccgacca agcgacgccc aaccggtatc agctcactca aaggcggtaa
     5101 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     5161 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     5221 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     5281 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     5341 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     5401 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     5461 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     5521 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     5581 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     5641 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     5701 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     5761 agattacgcg cagaaaaaaa ggatttcaag aagatccttt gatcttttct acggggtctg
     5821 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     5881 tcttcaccta gatcctttta tagtccggaa tcgtcacaca aaaaaccaac acacagatgt
     5941 aatgaaaata aagatatttt attgcggcca tcgtgatggc tagagtttag ccctcccaca
     6001 cataaccaga gggcagcaat tcacgaatcc caactgccgt cggctgtcca tcactgtcct
     6061 tcactatggc tttgatccca ggatgcagat cgagaagcac ctgtcggcac cgtccgcagg
     6121 ggctcaagat gcccctgttc tcatttccga tcgcgacgat acaagtcagg ttgccagctg
     6181 ccgcagcagc agcagtgccc agcaccacga gttctgcaca aggtccccca gtaaaatgat
     6241 atacattgac accagtgaag atgcggccgt cgctagagag agctgcgctg gcgacgctgt
     6301 agtcttcaga gatggggatg ctgttgattg tagccgttgc tctttcaatg agggtggatt
     6361 cttcttgaga caaaggcttg gccatggtgg cgctagtgtc agaagaatcg agctttttgc
     6421 aaaagcctag gcctccaaaa aagcctcctc actacttctg gaatagctca gaggccgagg
     6481 cggcctcggc ctctgcataa ataaaaaaaa ttagtcagcc atggggcgga gaatgggcgg
     6541 aactgggcgg agttaggggc gggatgggcg gagttagggg cgggactatg gttgctgact
     6601 aattgagatg catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc
     6661 tggttgctga ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg
     6721 actttccaca ccctaactga cacacattcc acagctggtt ctttccgcct cagaaggtac
     6781 ctaaccaagt tcctctttca gaggttattt caggccatgg tgctgcgc
//
