LOCUS       sequence-425880-        6309 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human NEDD1. C-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6309)
  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 6309)
  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_NEDD1:1-660
FEATURES             Location/Qualifiers
     source          1..6309
                     /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    3372..3493
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3383..3402)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(3608..3641)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3651..3698
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3730..4524
                     /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(3784..3803)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4394..4413
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4682..5270)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4762..4781)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5396..5444
                     /note="synthetic polyadenylation signal"
     CDS             complement(5468..5866)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5863..5872
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5897..6254)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5911..6046
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        6063..6254
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     6211..6231
                     /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 ccatgcagga aaacctcaga
     1261 tttgcttcat caggagatga tattaaaata tgggatgctt catctatgac attggtggat
     1321 aaattcaacc cacacacatc accacatgga atcagctcaa tatgttggag cagcaataat
     1381 aactttttag taacagcatc ttccagtggc gacaaaatag ttgtctcaag ttgcaaatgt
     1441 aaacctgttc cacttttaga gcttgctgaa gggcaaaagc agacatgtgt caatttaaat
     1501 tctacatcta tgtatttggt aagcggaggc ctaaataaca ctgttaatat ttgggattta
     1561 aaatcaaaaa gagttcatcg atctcttaag gatcataaag atcaagtaac ttgtgtaaca
     1621 tacaattgga atgattgcta cattgcttct ggatctctta gtggtgaaat tattttacac
     1681 agtgtaacca ctaatttatc tagtactcct tttggccatg gtagtaacca gtctgttcgg
     1741 cacttgaagt actccttgtt taagaaatca ctactgggca gtgtttcgga taatggaata
     1801 gtaactctct gggatgtaaa tagtcagagt ccataccata actttgacag tgtacacaaa
     1861 gctccagcgt caggcatctg tttttctcct gtcaatgaat tgctctttgt aaccataggc
     1921 ttggataaaa gaatcatcct ctatgacact tcaagtaaga agctagtgaa aactttagtg
     1981 gctgacactc ctctaactgc ggtagatttc atgcctgatg gagccacttt ggctattgga
     2041 tcttcccggg ggaaaatata tcaatatgat ttaagaatgt tgaaatcacc agttaagacc
     2101 atcagtgctc acaagacatc tgtgcagtgt atagcatttc agtactccac tgttcttact
     2161 aagtcaagtt taaataaagg ctgttcaaat aagcccacaa cagtgaacaa acgaagtgtt
     2221 aatgtgaatg ctgctagtgg aggagttcag aattccggaa ttgtcagaga agcacctgcc
     2281 acgtccattg ccacagttct accacaacct atgacatcag ctatggggaa aggaacagtt
     2341 gctgttcaag aaaaagcagg tttgcctcga agcataaaca cagacacttt atctaaggaa
     2401 acagacagtg gaaaaaatca ggatttctcc agctttgatg atactgggaa aagtagttta
     2461 ggtgacatgt tctcacctat cagagatgat gctgtagtta acaagggaag tgatgagtcc
     2521 ataggcaaag gagatggctt tgactttcta ccgcagttga actcagtgtt tcctccaaga
     2581 aaaaatccag taacttcaag tacttcagta ttgcattcta gtcctcttaa tgtttttatg
     2641 ggatctccag ggaaagagga aaatgaaaac cgtgatctaa cagctgagtc taagaaaata
     2701 tatatgggaa aacaggaatc taaagactcc ttcaaacagt tagcaaagtt ggtcacatct
     2761 ggtgctgaaa gtggaaatct aaatacctct ccatcatcta accaaacaag aaattctgag
     2821 aaatttgaaa agccagagaa tgaaattgaa gcccagttga tatgtgaacc cccaatcaat
     2881 ggatcctcaa ctccaaatcc aaagatagca tcttctgtca ctgctggagt tgccagttca
     2941 ctctcagaaa aaatagccga cagcattgga aataaccggc aaaatgcacc attgacttcc
     3001 attcaaattc gttttattca gaacatgata caggaaacgt tggatgactt tagagaagca
     3061 tgccataggg acattgtgaa tttgcaagtg gagatgatta aacagtttca tatgcaactg
     3121 aatgaaatgc attctttgct ggaaagatac tcagtgaatg aaggtttagt ggctgaaatt
     3181 gaaagactac gagaagaaaa caaaagatta cgggcccact tttggctcga gcggtgggag
     3241 ctccggtgtg agcggctggc ggctgttcaa gaagattagc taagtttaaa cggccgcgac
     3301 tctagactgc aggcatgcaa gctgatttgg ctgctgccac cgctgagcaa ggccgcttcg
     3361 agcagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     3421 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     3481 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggagat
     3541 gtgggaggtt tttttaagca agtaaaacct ctacaaatgt ggtaaaatct taattaagac
     3601 ttctaatata acttcgtata gcatacatta tacgaagtta tggttccact gttgacaatt
     3661 aatcatcggc atagtatatc ggcatagtat aatacgacaa ggtgaggaac taaacccagg
     3721 aggcagatca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     3781 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     3841 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     3901 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     3961 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     4021 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     4081 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     4141 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     4201 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     4261 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     4321 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     4381 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     4441 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     4501 cgccttcttg acgagttctt ctgattcgaa atgaccgacc aagcgacgcc caaccggtat
     4561 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     4621 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     4681 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     4741 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     4801 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     4861 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     4921 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     4981 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     5041 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     5101 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     5161 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     5221 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatttcaa gaagatcctt
     5281 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     5341 tcatgagatt atcaaaaagg atcttcacct agatcctttt atagtccgga atcgtcacac
     5401 aaaaaaccaa cacacagatg taatgaaaat aaagatattt tattgcggcc atcgtgatgg
     5461 ctagagttta gccctcccac acataaccag agggcagcaa ttcacgaatc ccaactgccg
     5521 tcggctgtcc atcactgtcc ttcactatgg ctttgatccc aggatgcaga tcgagaagca
     5581 cctgtcggca ccgtccgcag gggctcaaga tgcccctgtt ctcatttccg atcgcgacga
     5641 tacaagtcag gttgccagct gccgcagcag cagcagtgcc cagcaccacg agttctgcac
     5701 aaggtccccc agtaaaatga tatacattga caccagtgaa gatgcggccg tcgctagaga
     5761 gagctgcgct ggcgacgctg tagtcttcag agatggggat gctgttgatt gtagccgttg
     5821 ctctttcaat gagggtggat tcttcttgag acaaaggctt ggccatggtg gcgctagtgt
     5881 cagaagaatc gagctttttg caaaagccta ggcctccaaa aaagcctcct cactacttct
     5941 ggaatagctc agaggccgag gcggcctcgg cctctgcata aataaaaaaa attagtcagc
     6001 catggggcgg agaatgggcg gaactgggcg gagttagggg cgggatgggc ggagttaggg
     6061 gcgggactat ggttgctgac taattgagat gcatgctttg catacttctg cctgctgggg
     6121 agcctgggga ctttccacac ctggttgctg actaattgag atgcatgctt tgcatacttc
     6181 tgcctgctgg ggagcctggg gactttccac accctaactg acacacattc cacagctggt
     6241 tctttccgcc tcagaaggta cctaaccaag ttcctctttc agaggttatt tcaggccatg
     6301 gtgctgcgc
//
