LOCUS       sequence-426022-        6265 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human NEDD1. N-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6265)
  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 6265)
  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_NEDD1:1-660
FEATURES             Location/Qualifiers
     source          1..6265
                     /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    3328..3449
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3339..3358)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(3564..3597)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3607..3654
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3686..4480
                     /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(3740..3759)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4350..4369
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4638..5226)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4718..4737)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5352..5400
                     /note="synthetic polyadenylation signal"
     CDS             complement(5424..5822)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5819..5828
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5853..6210)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5867..6002
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        6019..6210
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     6167..6187
                     /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 tgcaggaaaa
     1261 cctcagattt gcttcatcag gagatgatat taaaatatgg gatgcttcat ctatgacatt
     1321 ggtggataaa ttcaacccac acacatcacc acatggaatc agctcaatat gttggagcag
     1381 caataataac tttttagtaa cagcatcttc cagtggcgac aaaatagttg tctcaagttg
     1441 caaatgtaaa cctgttccac ttttagagct tgctgaaggg caaaagcaga catgtgtcaa
     1501 tttaaattct acatctatgt atttggtaag cggaggccta aataacactg ttaatatttg
     1561 ggatttaaaa tcaaaaagag ttcatcgatc tcttaaggat cataaagatc aagtaacttg
     1621 tgtaacatac aattggaatg attgctacat tgcttctgga tctcttagtg gtgaaattat
     1681 tttacacagt gtaaccacta atttatctag tactcctttt ggccatggta gtaaccagtc
     1741 tgttcggcac ttgaagtact ccttgtttaa gaaatcacta ctgggcagtg tttcggataa
     1801 tggaatagta actctctggg atgtaaatag tcagagtcca taccataact ttgacagtgt
     1861 acacaaagct ccagcgtcag gcatctgttt ttctcctgtc aatgaattgc tctttgtaac
     1921 cataggcttg gataaaagaa tcatcctcta tgacacttca agtaagaagc tagtgaaaac
     1981 tttagtggct gacactcctc taactgcggt agatttcatg cctgatggag ccactttggc
     2041 tattggatct tcccggggga aaatatatca atatgattta agaatgttga aatcaccagt
     2101 taagaccatc agtgctcaca agacatctgt gcagtgtata gcatttcagt actccactgt
     2161 tcttactaag tcaagtttaa ataaaggctg ttcaaataag cccacaacag tgaacaaacg
     2221 aagtgttaat gtgaatgctg ctagtggagg agttcagaat tccggaattg tcagagaagc
     2281 acctgccacg tccattgcca cagttctacc acaacctatg acatcagcta tggggaaagg
     2341 aacagttgct gttcaagaaa aagcaggttt gcctcgaagc ataaacacag acactttatc
     2401 taaggaaaca gacagtggaa aaaatcagga tttctccagc tttgatgata ctgggaaaag
     2461 tagtttaggt gacatgttct cacctatcag agatgatgct gtagttaaca agggaagtga
     2521 tgagtccata ggcaaaggag atggctttga ctttctaccg cagttgaact cagtgtttcc
     2581 tccaagaaaa aatccagtaa cttcaagtac ttcagtattg cattctagtc ctcttaatgt
     2641 ttttatggga tctccaggga aagaggaaaa tgaaaaccgt gatctaacag ctgagtctaa
     2701 gaaaatatat atgggaaaac aggaatctaa agactccttc aaacagttag caaagttggt
     2761 cacatctggt gctgaaagtg gaaatctaaa tacctctcca tcatctaacc aaacaagaaa
     2821 ttctgagaaa tttgaaaagc cagagaatga aattgaagcc cagttgatat gtgaaccccc
     2881 aatcaatgga tcctcaactc caaatccaaa gatagcatct tctgtcactg ctggagttgc
     2941 cagttcactc tcagaaaaaa tagccgacag cattggaaat aaccggcaaa atgcaccatt
     3001 gacttccatt caaattcgtt ttattcagaa catgatacag gaaacgttgg atgactttag
     3061 agaagcatgc catagggaca ttgtgaattt gcaagtggag atgattaaac agtttcatat
     3121 gcaactgaat gaaatgcatt ctttgctgga aagatactca gtgaatgaag gtttagtggc
     3181 tgaaattgaa agactacgag aagaaaacaa aagattacgg gcccactttt taatctagag
     3241 gtcaagcttc gggggtttaa acctgcaggc atgcaagctg atttggctgc tgccaccgct
     3301 gagcaaggcc gcttcgagca gacatgataa gatacattga tgagtttgga caaaccacaa
     3361 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
     3421 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
     3481 aggttcaggg ggagatgtgg gaggtttttt taagcaagta aaacctctac aaatgtggta
     3541 aaatcttaat taagacttct aatataactt cgtatagcat acattatacg aagttatggt
     3601 tccactgttg acaattaatc atcggcatag tatatcggca tagtataata cgacaaggtg
     3661 aggaactaaa cccaggaggc agatcatgat tgaacaagat ggattgcacg caggttctcc
     3721 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     3781 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     3841 cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac
     3901 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     3961 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     4021 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     4081 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     4141 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     4201 caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     4261 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     4321 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     4381 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     4441 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga ttcgaaatga ccgaccaagc
     4501 gacgcccaac cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     4561 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     4621 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     4681 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     4741 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     4801 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     4861 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     4921 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     4981 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     5041 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
     5101 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     5161 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     5221 tttcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     5281 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttatag
     5341 tccggaatcg tcacacaaaa aaccaacaca cagatgtaat gaaaataaag atattttatt
     5401 gcggccatcg tgatggctag agtttagccc tcccacacat aaccagaggg cagcaattca
     5461 cgaatcccaa ctgccgtcgg ctgtccatca ctgtccttca ctatggcttt gatcccagga
     5521 tgcagatcga gaagcacctg tcggcaccgt ccgcaggggc tcaagatgcc cctgttctca
     5581 tttccgatcg cgacgataca agtcaggttg ccagctgccg cagcagcagc agtgcccagc
     5641 accacgagtt ctgcacaagg tcccccagta aaatgatata cattgacacc agtgaagatg
     5701 cggccgtcgc tagagagagc tgcgctggcg acgctgtagt cttcagagat ggggatgctg
     5761 ttgattgtag ccgttgctct ttcaatgagg gtggattctt cttgagacaa aggcttggcc
     5821 atggtggcgc tagtgtcaga agaatcgagc tttttgcaaa agcctaggcc tccaaaaaag
     5881 cctcctcact acttctggaa tagctcagag gccgaggcgg cctcggcctc tgcataaata
     5941 aaaaaaatta gtcagccatg gggcggagaa tgggcggaac tgggcggagt taggggcggg
     6001 atgggcggag ttaggggcgg gactatggtt gctgactaat tgagatgcat gctttgcata
     6061 cttctgcctg ctggggagcc tggggacttt ccacacctgg ttgctgacta attgagatgc
     6121 atgctttgca tacttctgcc tgctggggag cctggggact ttccacaccc taactgacac
     6181 acattccaca gctggttctt tccgcctcag aaggtaccta accaagttcc tctttcagag
     6241 gttatttcag gccatggtgc tgcgc
//
