LOCUS       sequence-423450-        7043 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human SCAP isoform 2. C-terminal
            HiBiT tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7043)
  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 7043)
  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_SCAP:1-905
FEATURES             Location/Qualifiers
     source          1..7043
                     /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    4106..4227
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4117..4136)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(4342..4375)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        4385..4432
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4464..5258
                     /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(4518..4537)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5128..5147
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(5416..6004)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5496..5515)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    6130..6178
                     /note="synthetic polyadenylation signal"
     CDS             complement(6202..6600)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      6597..6606
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(6631..6988)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6645..6780
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        6797..6988
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     6945..6965
                     /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 ccatgaccct gactgaaagg
     1261 ctgcgtgaga agatatctcg ggccttctac aaccatgggc tcctctgtgc atcctatccc
     1321 atccccatca tcctcatcgg ctacttcacc ctagtgcccg ccatccagga gttctgtctc
     1381 tttgctgtcg tggggctggt gtctgacttc ttccttcaga tgctgttttt caccactgtc
     1441 ctgtccattg acattcgccg gatggagcta gcagacctga acaagcgact gccccctgag
     1501 gcctgcctgc cctcagccaa gccagtgggg cagccaacgc gctacgagcg gcagctggct
     1561 gtgaggccgt ccacacccca caccatcacg ttgcagccgt cttccttccg aaacctgcgg
     1621 ctccccaaga ggctgcgtgt tgtctacttc ctggcccgca cccgcctggc acagcgcctc
     1681 atcatggctg gcaccgttgt ctggattggc atcctggtat acacagaccc agcagggctg
     1741 cgcaactacc tcgctgccca ggtgacggaa cagagcccat tgggtgaggg agccctggct
     1801 cccatgcccg tgcctagtgg catgctgccc cccagccacc cggaccctgc cttctccatc
     1861 ttcccacctg atgcccctaa gctacctgag aaccagacgt cgccaggcga gtcacctgag
     1921 cgtggaggtc cagcagaggt tgtccatgac agcccagtcc cagaggtaac ctgggggcct
     1981 gaggatgagg aactttggag gaaattgtcc ttccgccact ggccgacgct cttcagctat
     2041 tacaacatca cactggccaa gaggtacatc agcctgctgc ccgtcatccc agtcacgctc
     2101 cgcctgaacc cgagggaggc tctggagggc cggcaccctc aggacggccg cagtgcctgg
     2161 cccccaccgg ggcccatacc tgctgggcac tgggaagcag gacccaaggg cccaggtggg
     2221 gtgcaggccc atggagacgt cacgctgtac aaggtggcgg cgctgggcct ggccaccggc
     2281 atcgtcttgg tgctgctgct gctctgcctc taccgcgtgc tatgcccgcg caactacggg
     2341 cagctgggtg gtgggcccgg gcggcggagg cgcggggagc tgccctgcga cgactacggc
     2401 tatgcgccac ccgagacgga gatcgtgccg cttgtgctgc gcggccacct catggacatc
     2461 gagtgcctgg ccagcgacgg catgctgctg gtgagctgct gcctggcagg ccacatctgc
     2521 gtgtgggacg cgcagaccgg ggattgccta acgcgcattc cgcgcccagg gcagcgccgg
     2581 gacagtggcg tgggcagcgg gcttgaggct caggagagct gggaacgact ttcagatggt
     2641 gggaaggctg gtccagagga gcctggggac agccctcccc tgagacaccg cccccggggc
     2701 cctccgccgc cttccctctt cggggaccag cctgacctca cctgcttaat tgacaccaac
     2761 ttttcagcgc agcctcggtc ctcacagccc actcagcccg agccccggca ccgggcggtc
     2821 tgtggccgct ctcgggactc cccaggctat gacttcagct gcctggtgca gcgggtgtac
     2881 caggaggagg ggctggcggc cgtctgcaca ccagccctgc gcccaccctc gcctgggccg
     2941 gtgctgtccc aggcccctga ggacgagggt ggctcccccg agaaaggctc cccttccctc
     3001 gcctgggccc ccagtgccga gggttccatc tggagcttgg agctgcaggg caacctcatc
     3061 gtggtggggc ggagcagcgg ccggctggag gtgtgggacg ccattgaagg ggtgctgtgc
     3121 tgcagcagcg aggaggtctc ctcaggcatt accgctctgg tgttcttgga caaaaggatt
     3181 gtggctgcac ggctcaacgg ttcccttgat ttcttctcct tggagaccca cactgccctc
     3241 agccccctgc agtttagagg gaccccaggg cggggcagtt cccctgcctc tccagtgtac
     3301 agcagcagcg acacagtggc ctgtcacctg acccacacag tgccctgtgc acaccaaaaa
     3361 cccatcacag ccctgaaagc cgctgctggg cgcttggtga ctgggagcca agaccacaca
     3421 ctgagagtgt tccgtctgga ggactcgtgc tgcctcttca cccttcaggg ccactcaggg
     3481 gccatcacga ccgtgtacat tgaccagacc atggtgctgg ccagtggagg acaagatggg
     3541 gccatctgcc tgtgggatgt actgactggc agccgggtca gccatgtgtt tgctcaccgt
     3601 ggggatgtca cctcccttac ctgtaccacc tcctgtgtca tcagcagtgg cctggatgac
     3661 ctcatcagca tctgggaccg cagcacaggc atcaagttct actccattca gcaggacctg
     3721 ggctgtggtg caagcttggg tgtcatctca gacaacctgc tggtgactgg cggccagggc
     3781 tgtgtctcct tttgggacct aaactacggg gacctgttac agacagtcta cctggggaag
     3841 aacagtgagg cccagcctgc ccgccagatc ctggtgctgg acaacgctgc cattgtctgc
     3901 aactttggca gtgagctcag cctggtgtat gtgccctctg tgctggagaa gctggacggc
     3961 tcgagcggtg ggagctccgg tgtgagcggc tggcggctgt tcaagaagat tagctaagtt
     4021 taaacggccg cgactctaga ctgcaggcat gcaagctgat ttggctgctg ccaccgctga
     4081 gcaaggccgc ttcgagcaga catgataaga tacattgatg agtttggaca aaccacaact
     4141 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
     4201 accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag
     4261 gttcaggggg agatgtggga ggttttttta agcaagtaaa acctctacaa atgtggtaaa
     4321 atcttaatta agacttctaa tataacttcg tatagcatac attatacgaa gttatggttc
     4381 cactgttgac aattaatcat cggcatagta tatcggcata gtataatacg acaaggtgag
     4441 gaactaaacc caggaggcag atcatgattg aacaagatgg attgcacgca ggttctccgg
     4501 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     4561 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     4621 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
     4681 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     4741 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     4801 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     4861 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     4921 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     4981 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     5041 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     5101 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     5161 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     5221 gcatcgcctt ctatcgcctt cttgacgagt tcttctgatt cgaaatgacc gaccaagcga
     5281 cgcccaaccg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     5341 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     5401 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     5461 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     5521 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     5581 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     5641 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     5701 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     5761 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     5821 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
     5881 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     5941 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatt
     6001 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     6061 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttatagtc
     6121 cggaatcgtc acacaaaaaa ccaacacaca gatgtaatga aaataaagat attttattgc
     6181 ggccatcgtg atggctagag tttagccctc ccacacataa ccagagggca gcaattcacg
     6241 aatcccaact gccgtcggct gtccatcact gtccttcact atggctttga tcccaggatg
     6301 cagatcgaga agcacctgtc ggcaccgtcc gcaggggctc aagatgcccc tgttctcatt
     6361 tccgatcgcg acgatacaag tcaggttgcc agctgccgca gcagcagcag tgcccagcac
     6421 cacgagttct gcacaaggtc ccccagtaaa atgatataca ttgacaccag tgaagatgcg
     6481 gccgtcgcta gagagagctg cgctggcgac gctgtagtct tcagagatgg ggatgctgtt
     6541 gattgtagcc gttgctcttt caatgagggt ggattcttct tgagacaaag gcttggccat
     6601 ggtggcgcta gtgtcagaag aatcgagctt tttgcaaaag cctaggcctc caaaaaagcc
     6661 tcctcactac ttctggaata gctcagaggc cgaggcggcc tcggcctctg cataaataaa
     6721 aaaaattagt cagccatggg gcggagaatg ggcggaactg ggcggagtta ggggcgggat
     6781 gggcggagtt aggggcggga ctatggttgc tgactaattg agatgcatgc tttgcatact
     6841 tctgcctgct ggggagcctg gggactttcc acacctggtt gctgactaat tgagatgcat
     6901 gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacccta actgacacac
     6961 attccacagc tggttctttc cgcctcagaa ggtacctaac caagttcctc tttcagaggt
     7021 tatttcaggc catggtgctg cgc
//
