LOCUS       sequence-423457-        5621 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human CSTF1. C-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5621)
  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 5621)
  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_CSTF1:1-431
FEATURES             Location/Qualifiers
     source          1..5621
                     /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    2684..2805
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2695..2714)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(2920..2953)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2963..3010
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3042..3836
                     /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(3096..3115)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3706..3725
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(3994..4582)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4074..4093)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4708..4756
                     /note="synthetic polyadenylation signal"
     CDS             complement(4780..5178)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5175..5184
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5209..5566)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5223..5358
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5375..5566
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5523..5543
                     /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 ccatgtacag aaccaaagtg
     1261 ggcttgaagg accgccagca gctctacaag ctgatcatta gccagctgct atatgacggc
     1321 tacatcagca tcgccaatgg cctcatcaat gaaatcaagc ctcagtctgt gtgtgcaccc
     1381 tcggagcagc tcctgcatct catcaaactc ggaatggaaa acgatgacac cgcagttcag
     1441 tatgcaattg gtcgttcaga tactgttgcc cctggcacag ggattgacct ggaatttgat
     1501 gcagatgttc agactatgtc cccagaggct tctgagtacg aaacatgcta tgtcacatca
     1561 cataaaggac catgccgtgt agctacctat agtagagatg gacagttaat agctactggg
     1621 tctgctgatg cttcgataaa gatacttgac acagagagga tgttggccaa aagtgccatg
     1681 ccaatagagg tcatgatgaa tgagaccgca caacaaaata tggaaaacca cccagtgatt
     1741 cgaactcttt atgaccatgt ggatgaagtc acgtgccttg ctttccaccc aacagaacag
     1801 atcctggctt ctggttcaag ggattatact cttaaattat ttgattattc caaaccatca
     1861 gcaaaaagag ccttcaaata cattcaggaa gctgaaatgt tacgttccat ctcttttcat
     1921 ccttctggag actttatact tgtcggaact cagcatccta ctcttcgcct ttatgatatc
     1981 aacacctttc aatgttttgt ctcttgcaat cctcaagatc aacacaccga tgctatatgt
     2041 tccgttaatt acaattctag tgccaatatg tacgtaactg gaagcaagga cggctgcatc
     2101 aaattatggg atggtgtttc aaatcgatgc atcacaactt ttgagaaagc acatgacggt
     2161 gctgaagttt gttctgccat tttttccaaa aattctaaat acattctctc aagtggaaaa
     2221 gactctgtag ctaaactttg ggaaatatca acgggacgaa cactggtcag atacacgggc
     2281 gcgggtttaa gtggacgcca ggtgcaccgg acacaggctg tgtttaacca caccgaggac
     2341 tatgtgttgc tgcccgacga gaggacgatc agtctttgct gctgggactc gaggacagcc
     2401 gagcggagaa acctgctgtc gttggggcac aacaatattg tacgctgcat agtgcactcc
     2461 cccaccaacc ccgggttcat gacgtgcagc gatgacttca gagcgcggtt ttggtaccgg
     2521 agatcgacca ctgacggctc gagcggtggg agctccggtg tgagcggctg gcggctgttc
     2581 aagaagatta gctaagttta aacggccgcg actctagact gcaggcatgc aagctgattt
     2641 ggctgctgcc accgctgagc aaggccgctt cgagcagaca tgataagata cattgatgag
     2701 tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat
     2761 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
     2821 attcatttta tgtttcaggt tcagggggag atgtgggagg tttttttaag caagtaaaac
     2881 ctctacaaat gtggtaaaat cttaattaag acttctaata taacttcgta tagcatacat
     2941 tatacgaagt tatggttcca ctgttgacaa ttaatcatcg gcatagtata tcggcatagt
     3001 ataatacgac aaggtgagga actaaaccca ggaggcagat catgattgaa caagatggat
     3061 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     3121 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     3181 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc
     3241 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     3301 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     3361 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     3421 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     3481 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     3541 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga
     3601 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     3661 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     3721 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     3781 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgattcg
     3841 aaatgaccga ccaagcgacg cccaaccggt atcagctcac tcaaaggcgg taatacggtt
     3901 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     3961 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     4021 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     4081 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     4141 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     4201 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     4261 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     4321 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     4381 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     4441 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     4501 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     4561 gcgcagaaaa aaaggatttc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     4621 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     4681 ctagatcctt ttatagtccg gaatcgtcac acaaaaaacc aacacacaga tgtaatgaaa
     4741 ataaagatat tttattgcgg ccatcgtgat ggctagagtt tagccctccc acacataacc
     4801 agagggcagc aattcacgaa tcccaactgc cgtcggctgt ccatcactgt ccttcactat
     4861 ggctttgatc ccaggatgca gatcgagaag cacctgtcgg caccgtccgc aggggctcaa
     4921 gatgcccctg ttctcatttc cgatcgcgac gatacaagtc aggttgccag ctgccgcagc
     4981 agcagcagtg cccagcacca cgagttctgc acaaggtccc ccagtaaaat gatatacatt
     5041 gacaccagtg aagatgcggc cgtcgctaga gagagctgcg ctggcgacgc tgtagtcttc
     5101 agagatgggg atgctgttga ttgtagccgt tgctctttca atgagggtgg attcttcttg
     5161 agacaaaggc ttggccatgg tggcgctagt gtcagaagaa tcgagctttt tgcaaaagcc
     5221 taggcctcca aaaaagcctc ctcactactt ctggaatagc tcagaggccg aggcggcctc
     5281 ggcctctgca taaataaaaa aaattagtca gccatggggc ggagaatggg cggaactggg
     5341 cggagttagg ggcgggatgg gcggagttag gggcgggact atggttgctg actaattgag
     5401 atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac acctggttgc
     5461 tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc
     5521 acaccctaac tgacacacat tccacagctg gttctttccg cctcagaagg tacctaacca
     5581 agttcctctt tcagaggtta tttcaggcca tggtgctgcg c
//
