LOCUS       sequence-424668-        6584 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human CDRT1. C-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6584)
  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 6584)
  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_CDRT1:1-752
FEATURES             Location/Qualifiers
     source          1..6584
                     /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"
     regulatory      2167..2176
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3647..3768
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3658..3677)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(3883..3916)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3926..3973
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4005..4799
                     /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(4059..4078)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4669..4688
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4957..5545)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5037..5056)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5671..5719
                     /note="synthetic polyadenylation signal"
     CDS             complement(5743..6141)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      6138..6147
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(6172..6529)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6186..6321
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        6338..6529
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     6486..6506
                     /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 ccatggaaaa cctggaatca
     1261 aggctcaaga atgcccccta ttttcgttgt gagaagggaa ccgattccat ccctctatgc
     1321 cggaagtgtg agacgcgtgt cttagcctgg aagatcttct ctaccaaaga gtggttctgc
     1381 aggatcaatg acatatcaca gaggaggttt ctagttggca ttctgaagca gttaaatagc
     1441 ttatatttgt tacactattt ccaaaatatc cttcagacca cacagggaaa ggatttcatc
     1501 tataacaggt cccggatcga cctcagcaag aaagagggga aagttgtgaa gtcatccttg
     1561 aaccaaatgt tggataaaac agtagaacag aagatgaaag agatcttgta ttggtttgcg
     1621 aacagcaccc agtggaccaa ggcgaattat actctcttac tgctgcagat gtgcaacccc
     1681 aaattactgc tcactgctgc caatgtgatc agagtcctgt ttctgagaga ggagaacaat
     1741 atctcagggc tcaatcaaga catcacagat gtgtgttttt cccctgagaa agaccacagc
     1801 tccaagtctg cgacctcaca agtctattgg acagccaaaa ctcagcacac atcccttcct
     1861 ttgtccaaag ccccagaaaa tgaacacttc cttggggcag catctaaccc tgaggaacca
     1921 tggaggaatt cactccggtg tatatccgaa atgaataggc tgttttctgg aaaagcagac
     1981 ataaccaagc cagggtacga tccctgcaat ctattggttg acctggatga catcagagac
     2041 ctgtcttctg ggttcagcaa ataccgagac ttcatccgtt acctgcccat ccacctctcc
     2101 aagtacattc taagaatgct ggatagacac accctgaaca agtgcgcctc tgtgagccag
     2161 cactgggccg ccatggctca acaggtcaag atggacttgt cagcgcacgg cttcattcag
     2221 aaccagatta ccttcttgca ggggtcctat acaagaggaa ttgatcctaa ttatgccaat
     2281 aaggtttcta tcccagttcc taaaatggta gatgacggga agagcatgcg tgtgaaacat
     2341 ccgaagtgga agttgagaac gaagaatgag tacaacctgt ggactgcata ccagaacgag
     2401 gaaacgcagc aggtcctgat ggaggagaga aatgttttct gtgggaccta caatgttcgc
     2461 attctctctg acacgtggga tcaaaaccga gtcatccact attccggggg agatctgata
     2521 gctgtgtcat ctaatcgaaa gatccatctt ctggacatca tacaagtgaa agcgataccc
     2581 gttgaattcc gaggccatgc tgggagtgtc cgggccctct tcctgtgtga ggaggaaaac
     2641 tttctcctaa gcgggagcta tgacctaagt atcagatact gggatctgaa aagtggggtt
     2701 tgcacacgaa tcttcggtgg tcaccagggg actatcactt gcatggactt gtgtaagaac
     2761 aggctcgtat ctggaggaag agattgccag gtaaaagtat gggatgtaga cacagggaag
     2821 tgcctgaaga cgtttagaca caaagacccc atcttggcca ccaggatcaa tgatacctac
     2881 attgtgagca gctgtgagcg agggctggtg aaagtgtggc acattgccat ggcccagttg
     2941 gtaaagactc tcagtggcca cgagggagct gtgaaatgcc tgttctttga ccagtggcat
     3001 ctcctctcag gaagcactga tggcctggtc atggcctgga gcatggtggg gaagtacgag
     3061 cgctgcctga tggccttcaa gcatcccaaa gaggtgctcg acgtgtccct tctcttcctc
     3121 cgggtcatca gcgcctgtgc agatggcaag atccgaattt acaatttctt caatgggaac
     3181 tgtatgaagg tgataaaagc caatggcaga ggcgatcctg tgctgtcctt ctttattcag
     3241 ggcaacagaa tttcagtctg ccacatcagc acatttgcta aaagaattaa cgtgggatgg
     3301 aatggaatcg aaccaagtgc tacagctcag ggaggaaatg cctccttgac cgagtgtgct
     3361 catgtgagac tccacatcgc aggacactta ccagcatcga ggctgcccgt ggccgctgtc
     3421 cagcccatga caggcgggat ggccccaacc acagctccga cccatgtgtt ggcaatgctg
     3481 atccttttca gtggtgtggg ctcgagcggt gggagctccg gtgtgagcgg ctggcggctg
     3541 ttcaagaaga ttagctaagt ttaaacggcc gcgactctag actgcaggca tgcaagctga
     3601 tttggctgct gccaccgctg agcaaggccg cttcgagcag acatgataag atacattgat
     3661 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt
     3721 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat
     3781 tgcattcatt ttatgtttca ggttcagggg gagatgtggg aggttttttt aagcaagtaa
     3841 aacctctaca aatgtggtaa aatcttaatt aagacttcta atataacttc gtatagcata
     3901 cattatacga agttatggtt ccactgttga caattaatca tcggcatagt atatcggcat
     3961 agtataatac gacaaggtga ggaactaaac ccaggaggca gatcatgatt gaacaagatg
     4021 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     4081 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     4141 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
     4201 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     4261 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     4321 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     4381 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     4441 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     4501 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg
     4561 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     4621 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     4681 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     4741 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgat
     4801 tcgaaatgac cgaccaagcg acgcccaacc ggtatcagct cactcaaagg cggtaatacg
     4861 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     4921 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     4981 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     5041 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     5101 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     5161 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     5221 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     5281 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     5341 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     5401 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     5461 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     5521 tacgcgcaga aaaaaaggat ttcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     5581 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     5641 cacctagatc cttttatagt ccggaatcgt cacacaaaaa accaacacac agatgtaatg
     5701 aaaataaaga tattttattg cggccatcgt gatggctaga gtttagccct cccacacata
     5761 accagagggc agcaattcac gaatcccaac tgccgtcggc tgtccatcac tgtccttcac
     5821 tatggctttg atcccaggat gcagatcgag aagcacctgt cggcaccgtc cgcaggggct
     5881 caagatgccc ctgttctcat ttccgatcgc gacgatacaa gtcaggttgc cagctgccgc
     5941 agcagcagca gtgcccagca ccacgagttc tgcacaaggt cccccagtaa aatgatatac
     6001 attgacacca gtgaagatgc ggccgtcgct agagagagct gcgctggcga cgctgtagtc
     6061 ttcagagatg gggatgctgt tgattgtagc cgttgctctt tcaatgaggg tggattcttc
     6121 ttgagacaaa ggcttggcca tggtggcgct agtgtcagaa gaatcgagct ttttgcaaaa
     6181 gcctaggcct ccaaaaaagc ctcctcacta cttctggaat agctcagagg ccgaggcggc
     6241 ctcggcctct gcataaataa aaaaaattag tcagccatgg ggcggagaat gggcggaact
     6301 gggcggagtt aggggcggga tgggcggagt taggggcggg actatggttg ctgactaatt
     6361 gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc cacacctggt
     6421 tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt
     6481 tccacaccct aactgacaca cattccacag ctggttcttt ccgcctcaga aggtacctaa
     6541 ccaagttcct ctttcagagg ttatttcagg ccatggtgct gcgc
//
