LOCUS       sequence-425922-        6999 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human SCAP isoform 2. N-terminal
            HiBiT tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6999)
  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 6999)
  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_SCAP:1-905
FEATURES             Location/Qualifiers
     source          1..6999
                     /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    4062..4183
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4073..4092)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(4298..4331)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        4341..4388
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4420..5214
                     /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(4474..4493)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5084..5103
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(5372..5960)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5452..5471)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    6086..6134
                     /note="synthetic polyadenylation signal"
     CDS             complement(6158..6556)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      6553..6562
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(6587..6944)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6601..6736
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        6753..6944
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     6901..6921
                     /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 tgaccctgac
     1261 tgaaaggctg cgtgagaaga tatctcgggc cttctacaac catgggctcc tctgtgcatc
     1321 ctatcccatc cccatcatcc tcatcggcta cttcacccta gtgcccgcca tccaggagtt
     1381 ctgtctcttt gctgtcgtgg ggctggtgtc tgacttcttc cttcagatgc tgtttttcac
     1441 cactgtcctg tccattgaca ttcgccggat ggagctagca gacctgaaca agcgactgcc
     1501 ccctgaggcc tgcctgccct cagccaagcc agtggggcag ccaacgcgct acgagcggca
     1561 gctggctgtg aggccgtcca caccccacac catcacgttg cagccgtctt ccttccgaaa
     1621 cctgcggctc cccaagaggc tgcgtgttgt ctacttcctg gcccgcaccc gcctggcaca
     1681 gcgcctcatc atggctggca ccgttgtctg gattggcatc ctggtataca cagacccagc
     1741 agggctgcgc aactacctcg ctgcccaggt gacggaacag agcccattgg gtgagggagc
     1801 cctggctccc atgcccgtgc ctagtggcat gctgcccccc agccacccgg accctgcctt
     1861 ctccatcttc ccacctgatg cccctaagct acctgagaac cagacgtcgc caggcgagtc
     1921 acctgagcgt ggaggtccag cagaggttgt ccatgacagc ccagtcccag aggtaacctg
     1981 ggggcctgag gatgaggaac tttggaggaa attgtccttc cgccactggc cgacgctctt
     2041 cagctattac aacatcacac tggccaagag gtacatcagc ctgctgcccg tcatcccagt
     2101 cacgctccgc ctgaacccga gggaggctct ggagggccgg caccctcagg acggccgcag
     2161 tgcctggccc ccaccggggc ccatacctgc tgggcactgg gaagcaggac ccaagggccc
     2221 aggtggggtg caggcccatg gagacgtcac gctgtacaag gtggcggcgc tgggcctggc
     2281 caccggcatc gtcttggtgc tgctgctgct ctgcctctac cgcgtgctat gcccgcgcaa
     2341 ctacgggcag ctgggtggtg ggcccgggcg gcggaggcgc ggggagctgc cctgcgacga
     2401 ctacggctat gcgccacccg agacggagat cgtgccgctt gtgctgcgcg gccacctcat
     2461 ggacatcgag tgcctggcca gcgacggcat gctgctggtg agctgctgcc tggcaggcca
     2521 catctgcgtg tgggacgcgc agaccgggga ttgcctaacg cgcattccgc gcccagggca
     2581 gcgccgggac agtggcgtgg gcagcgggct tgaggctcag gagagctggg aacgactttc
     2641 agatggtggg aaggctggtc cagaggagcc tggggacagc cctcccctga gacaccgccc
     2701 ccggggccct ccgccgcctt ccctcttcgg ggaccagcct gacctcacct gcttaattga
     2761 caccaacttt tcagcgcagc ctcggtcctc acagcccact cagcccgagc cccggcaccg
     2821 ggcggtctgt ggccgctctc gggactcccc aggctatgac ttcagctgcc tggtgcagcg
     2881 ggtgtaccag gaggaggggc tggcggccgt ctgcacacca gccctgcgcc caccctcgcc
     2941 tgggccggtg ctgtcccagg cccctgagga cgagggtggc tcccccgaga aaggctcccc
     3001 ttccctcgcc tgggccccca gtgccgaggg ttccatctgg agcttggagc tgcagggcaa
     3061 cctcatcgtg gtggggcgga gcagcggccg gctggaggtg tgggacgcca ttgaaggggt
     3121 gctgtgctgc agcagcgagg aggtctcctc aggcattacc gctctggtgt tcttggacaa
     3181 aaggattgtg gctgcacggc tcaacggttc ccttgatttc ttctccttgg agacccacac
     3241 tgccctcagc cccctgcagt ttagagggac cccagggcgg ggcagttccc ctgcctctcc
     3301 agtgtacagc agcagcgaca cagtggcctg tcacctgacc cacacagtgc cctgtgcaca
     3361 ccaaaaaccc atcacagccc tgaaagccgc tgctgggcgc ttggtgactg ggagccaaga
     3421 ccacacactg agagtgttcc gtctggagga ctcgtgctgc ctcttcaccc ttcagggcca
     3481 ctcaggggcc atcacgaccg tgtacattga ccagaccatg gtgctggcca gtggaggaca
     3541 agatggggcc atctgcctgt gggatgtact gactggcagc cgggtcagcc atgtgtttgc
     3601 tcaccgtggg gatgtcacct cccttacctg taccacctcc tgtgtcatca gcagtggcct
     3661 ggatgacctc atcagcatct gggaccgcag cacaggcatc aagttctact ccattcagca
     3721 ggacctgggc tgtggtgcaa gcttgggtgt catctcagac aacctgctgg tgactggcgg
     3781 ccagggctgt gtctcctttt gggacctaaa ctacggggac ctgttacaga cagtctacct
     3841 ggggaagaac agtgaggccc agcctgcccg ccagatcctg gtgctggaca acgctgccat
     3901 tgtctgcaac tttggcagtg agctcagcct ggtgtatgtg ccctctgtgc tggagaagct
     3961 ggactaatct agaggtcaag cttcgggggt ttaaacctgc aggcatgcaa gctgatttgg
     4021 ctgctgccac cgctgagcaa ggccgcttcg agcagacatg ataagataca ttgatgagtt
     4081 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     4141 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     4201 tcattttatg tttcaggttc agggggagat gtgggaggtt tttttaagca agtaaaacct
     4261 ctacaaatgt ggtaaaatct taattaagac ttctaatata acttcgtata gcatacatta
     4321 tacgaagtta tggttccact gttgacaatt aatcatcggc atagtatatc ggcatagtat
     4381 aatacgacaa ggtgaggaac taaacccagg aggcagatca tgattgaaca agatggattg
     4441 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     4501 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     4561 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta
     4621 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     4681 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     4741 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     4801 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     4861 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     4921 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc
     4981 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     5041 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     5101 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     5161 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgattcgaa
     5221 atgaccgacc aagcgacgcc caaccggtat cagctcactc aaaggcggta atacggttat
     5281 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     5341 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     5401 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     5461 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     5521 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     5581 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     5641 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     5701 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     5761 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     5821 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     5881 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     5941 gcagaaaaaa aggatttcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     6001 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     6061 agatcctttt atagtccgga atcgtcacac aaaaaaccaa cacacagatg taatgaaaat
     6121 aaagatattt tattgcggcc atcgtgatgg ctagagttta gccctcccac acataaccag
     6181 agggcagcaa ttcacgaatc ccaactgccg tcggctgtcc atcactgtcc ttcactatgg
     6241 ctttgatccc aggatgcaga tcgagaagca cctgtcggca ccgtccgcag gggctcaaga
     6301 tgcccctgtt ctcatttccg atcgcgacga tacaagtcag gttgccagct gccgcagcag
     6361 cagcagtgcc cagcaccacg agttctgcac aaggtccccc agtaaaatga tatacattga
     6421 caccagtgaa gatgcggccg tcgctagaga gagctgcgct ggcgacgctg tagtcttcag
     6481 agatggggat gctgttgatt gtagccgttg ctctttcaat gagggtggat tcttcttgag
     6541 acaaaggctt ggccatggtg gcgctagtgt cagaagaatc gagctttttg caaaagccta
     6601 ggcctccaaa aaagcctcct cactacttct ggaatagctc agaggccgag gcggcctcgg
     6661 cctctgcata aataaaaaaa attagtcagc catggggcgg agaatgggcg gaactgggcg
     6721 gagttagggg cgggatgggc ggagttaggg gcgggactat ggttgctgac taattgagat
     6781 gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac ctggttgctg
     6841 actaattgag atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac
     6901 accctaactg acacacattc cacagctggt tctttccgcc tcagaaggta cctaaccaag
     6961 ttcctctttc agaggttatt tcaggccatg gtgctgcgc
//
