LOCUS       sequence-425791-        5235 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of human KIF21A fragment L1337 to A1653. 
            N-terminal HiBiT tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5235)
  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 5235)
  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_KIF21A:1337-1653
FEATURES             Location/Qualifiers
     source          1..5235
                     /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    2298..2419
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2309..2328)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(2534..2567)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2577..2624
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2656..3450
                     /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(2710..2729)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3320..3339
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(3608..4196)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3688..3707)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4322..4370
                     /note="synthetic polyadenylation signal"
     CDS             complement(4394..4792)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      4789..4798
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4823..5180)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4837..4972
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5137..5157
                     /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 tcgagcggtc ttcagtgtat
     1261 tcacatagct gaagggcata caaaagctgt gctctgtgtg gattctactg atgatctcct
     1321 cttcactgga tcaaaagatc gtacttgtaa agtatggaat ctggtgactg ggcaggaaat
     1381 aatgtcactg gggggtcatc ccaacaatgt cgtgtctgta aaatactgta attataccag
     1441 tttggtcttc actgtatcaa catcttatat taaggtgtgg gatatcagag attcagcaaa
     1501 gtgcattcga acactaacgt cttcaggtca agttactctt ggagatgctt gttctgcaag
     1561 taccagtcga acagtagcta ttccttctgg agagaaccag atcaatcaaa ttgccctaaa
     1621 cccaactggc accttcctct atgctgcttc tggaaatgct gtcaggatgt gggatcttaa
     1681 aaggtttcag tctacaggaa agttaacagg acacctaggc cctgttatgt gccttactgt
     1741 ggatcagatt tccagtggac aagatctaat catcactggc tccaaggatc attacatcaa
     1801 aatgtttgat gttacagaag gagctcttgg gactgtgagt cccacccaca attttgaacc
     1861 ccctcattat gatggcatag aagcactaac cattcaaggg gataacctat ttagtgggtc
     1921 tagagataat ggaatcaaga aatgggactt aactcaaaaa gaccttcttc agcaagttcc
     1981 aaatgcacat aaggattggg tctgtgccct gggagtggtg ccagaccacc cagttttgct
     2041 cagtggctgc agagggggca ttttgaaagt ctggaacatg gatactttta tgccagtggg
     2101 agagatgaag ggtcatgata gtcctatcaa tgctatatgt gttaattcca cccacatttt
     2161 tactgcagct gatgatcgaa ctgtgagaat ttggaaggct taatctagag gtcaagcttc
     2221 gggggtttaa acctgcaggc atgcaagctg atttggctgc tgccaccgct gagcaaggcc
     2281 gcttcgagca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca
     2341 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
     2401 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg
     2461 ggagatgtgg gaggtttttt taagcaagta aaacctctac aaatgtggta aaatcttaat
     2521 taagacttct aatataactt cgtatagcat acattatacg aagttatggt tccactgttg
     2581 acaattaatc atcggcatag tatatcggca tagtataata cgacaaggtg aggaactaaa
     2641 cccaggaggc agatcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     2701 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     2761 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     2821 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     2881 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     2941 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     3001 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     3061 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     3121 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     3181 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     3241 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     3301 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     3361 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     3421 ttctatcgcc ttcttgacga gttcttctga ttcgaaatga ccgaccaagc gacgcccaac
     3481 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     3541 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     3601 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     3661 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     3721 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     3781 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     3841 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     3901 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     3961 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     4021 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
     4081 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     4141 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tttcaagaag
     4201 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     4261 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttatag tccggaatcg
     4321 tcacacaaaa aaccaacaca cagatgtaat gaaaataaag atattttatt gcggccatcg
     4381 tgatggctag agtttagccc tcccacacat aaccagaggg cagcaattca cgaatcccaa
     4441 ctgccgtcgg ctgtccatca ctgtccttca ctatggcttt gatcccagga tgcagatcga
     4501 gaagcacctg tcggcaccgt ccgcaggggc tcaagatgcc cctgttctca tttccgatcg
     4561 cgacgataca agtcaggttg ccagctgccg cagcagcagc agtgcccagc accacgagtt
     4621 ctgcacaagg tcccccagta aaatgatata cattgacacc agtgaagatg cggccgtcgc
     4681 tagagagagc tgcgctggcg acgctgtagt cttcagagat ggggatgctg ttgattgtag
     4741 ccgttgctct ttcaatgagg gtggattctt cttgagacaa aggcttggcc atggtggcgc
     4801 tagtgtcaga agaatcgagc tttttgcaaa agcctaggcc tccaaaaaag cctcctcact
     4861 acttctggaa tagctcagag gccgaggcgg cctcggcctc tgcataaata aaaaaaatta
     4921 gtcagccatg gggcggagaa tgggcggaac tgggcggagt taggggcggg atgggcggag
     4981 ttaggggcgg gactatggtt gctgactaat tgagatgcat gctttgcata cttctgcctg
     5041 ctggggagcc tggggacttt csrmmymtgg ttgctgacta attgagatgc atgctttgca
     5101 tacttctgcc tgctggggag cctggggact ttccacaccc taactgacac acattccaca
     5161 gctggttctt tccgcctcag aaggtaccta accaagttcc tctttcagag gttatttcag
     5221 gccatggtgc tgcgc
//
