LOCUS       sequence-425930-        5577 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human CSTF1. N-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5577)
  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 5577)
  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_CSTF1:1-431
FEATURES             Location/Qualifiers
     source          1..5577
                     /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    2640..2761
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2651..2670)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(2876..2909)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2919..2966
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2998..3792
                     /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(3052..3071)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3662..3681
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(3950..4538)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4030..4049)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4664..4712
                     /note="synthetic polyadenylation signal"
     CDS             complement(4736..5134)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5131..5140
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5165..5522)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5179..5314
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5331..5522
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5479..5499
                     /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 tgtacagaac
     1261 caaagtgggc ttgaaggacc gccagcagct ctacaagctg atcattagcc agctgctata
     1321 tgacggctac atcagcatcg ccaatggcct catcaatgaa atcaagcctc agtctgtgtg
     1381 tgcaccctcg gagcagctcc tgcatctcat caaactcgga atggaaaacg atgacaccgc
     1441 agttcagtat gcaattggtc gttcagatac tgttgcccct ggcacaggga ttgacctgga
     1501 atttgatgca gatgttcaga ctatgtcccc agaggcttct gagtacgaaa catgctatgt
     1561 cacatcacat aaaggaccat gccgtgtagc tacctatagt agagatggac agttaatagc
     1621 tactgggtct gctgatgctt cgataaagat acttgacaca gagaggatgt tggccaaaag
     1681 tgccatgcca atagaggtca tgatgaatga gaccgcacaa caaaatatgg aaaaccaccc
     1741 agtgattcga actctttatg accatgtgga tgaagtcacg tgccttgctt tccacccaac
     1801 agaacagatc ctggcttctg gttcaaggga ttatactctt aaattatttg attattccaa
     1861 accatcagca aaaagagcct tcaaatacat tcaggaagct gaaatgttac gttccatctc
     1921 ttttcatcct tctggagact ttatacttgt cggaactcag catcctactc ttcgccttta
     1981 tgatatcaac acctttcaat gttttgtctc ttgcaatcct caagatcaac acaccgatgc
     2041 tatatgttcc gttaattaca attctagtgc caatatgtac gtaactggaa gcaaggacgg
     2101 ctgcatcaaa ttatgggatg gtgtttcaaa tcgatgcatc acaacttttg agaaagcaca
     2161 tgacggtgct gaagtttgtt ctgccatttt ttccaaaaat tctaaataca ttctctcaag
     2221 tggaaaagac tctgtagcta aactttggga aatatcaacg ggacgaacac tggtcagata
     2281 cacgggcgcg ggtttaagtg gacgccaggt gcaccggaca caggctgtgt ttaaccacac
     2341 cgaggactat gtgttgctgc ccgacgagag gacgatcagt ctttgctgct gggactcgag
     2401 gacagccgag cggagaaacc tgctgtcgtt ggggcacaac aatattgtac gctgcatagt
     2461 gcactccccc accaaccccg ggttcatgac gtgcagcgat gacttcagag cgcggttttg
     2521 gtaccggaga tcgaccactg actaatctag aggtcaagct tcgggggttt aaacctgcag
     2581 gcatgcaagc tgatttggct gctgccaccg ctgagcaagg ccgcttcgag cagacatgat
     2641 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     2701 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     2761 taacaacaac aattgcattc attttatgtt tcaggttcag ggggagatgt gggaggtttt
     2821 tttaagcaag taaaacctct acaaatgtgg taaaatctta attaagactt ctaatataac
     2881 ttcgtatagc atacattata cgaagttatg gttccactgt tgacaattaa tcatcggcat
     2941 agtatatcgg catagtataa tacgacaagg tgaggaacta aacccaggag gcagatcatg
     3001 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     3061 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     3121 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     3181 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     3241 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     3301 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     3361 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     3421 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     3481 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     3541 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     3601 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     3661 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     3721 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     3781 gagttcttct gattcgaaat gaccgaccaa gcgacgccca accggtatca gctcactcaa
     3841 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     3901 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     3961 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     4021 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     4081 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     4141 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     4201 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     4261 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     4321 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     4381 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     4441 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     4501 gcaagcagca gattacgcgc agaaaaaaag gatttcaaga agatcctttg atcttttcta
     4561 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     4621 caaaaaggat cttcacctag atccttttat agtccggaat cgtcacacaa aaaaccaaca
     4681 cacagatgta atgaaaataa agatatttta ttgcggccat cgtgatggct agagtttagc
     4741 cctcccacac ataaccagag ggcagcaatt cacgaatccc aactgccgtc ggctgtccat
     4801 cactgtcctt cactatggct ttgatcccag gatgcagatc gagaagcacc tgtcggcacc
     4861 gtccgcaggg gctcaagatg cccctgttct catttccgat cgcgacgata caagtcaggt
     4921 tgccagctgc cgcagcagca gcagtgccca gcaccacgag ttctgcacaa ggtcccccag
     4981 taaaatgata tacattgaca ccagtgaaga tgcggccgtc gctagagaga gctgcgctgg
     5041 cgacgctgta gtcttcagag atggggatgc tgttgattgt agccgttgct ctttcaatga
     5101 gggtggattc ttcttgagac aaaggcttgg ccatggtggc gctagtgtca gaagaatcga
     5161 gctttttgca aaagcctagg cctccaaaaa agcctcctca ctacttctgg aatagctcag
     5221 aggccgaggc ggcctcggcc tctgcataaa taaaaaaaat tagtcagcca tggggcggag
     5281 aatgggcgga actgggcgga gttaggggcg ggatgggcgg agttaggggc gggactatgg
     5341 ttgctgacta attgagatgc atgctttgca tacttctgcc tgctggggag cctggggact
     5401 ttccacacct ggttgctgac taattgagat gcatgctttg catacttctg cctgctgggg
     5461 agcctgggga ctttccacac cctaactgac acacattcca cagctggttc tttccgcctc
     5521 agaaggtacc taaccaagtt cctctttcag aggttatttc aggccatggt gctgcgc
//
