LOCUS       sequence-423435-        5282 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of human KIF21A fragment L1337 to A1653. 
            C-terminal HiBiT tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5282)
  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 5282)
  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_KIF21A:1337-1653
FEATURES             Location/Qualifiers
     source          1..5282
                     /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    2345..2466
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2356..2375)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(2581..2614)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2624..2671
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2703..3497
                     /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(2757..2776)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3367..3386
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(3655..4243)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3735..3754)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4369..4417
                     /note="synthetic polyadenylation signal"
     CDS             complement(4441..4839)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      4836..4845
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4870..5227)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4884..5019
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5036..5227
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5184..5204
                     /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 ccatgcttca gtgtattcac
     1261 atagctgaag ggcatacaaa agctgtgctc tgtgtggatt ctactgatga tctcctcttc
     1321 actggatcaa aagatcgtac ttgtaaagta tggaatctgg tgactgggca ggaaataatg
     1381 tcactggggg gtcatcccaa caatgtcgtg tctgtaaaat actgtaatta taccagtttg
     1441 gtcttcactg tatcaacatc ttatattaag gtgtgggata tcagagattc agcaaagtgc
     1501 attcgaacac taacgtcttc aggtcaagtt actcttggag atgcttgttc tgcaagtacc
     1561 agtcgaacag tagctattcc ttctggagag aaccagatca atcaaattgc cctaaaccca
     1621 actggcacct tcctctatgc tgcttctgga aatgctgtca ggatgtggga tcttaaaagg
     1681 tttcagtcta caggaaagtt aacaggacac ctaggccctg ttatgtgcct tactgtggat
     1741 cagatttcca gtggacaaga tctaatcatc actggctcca aggatcatta catcaaaatg
     1801 tttgatgtta cagaaggagc tcttgggact gtgagtccca cccacaattt tgaaccccct
     1861 cattatgatg gcatagaagc actaaccatt caaggggata acctatttag tgggtctaga
     1921 gataatggaa tcaagaaatg ggacttaact caaaaagacc ttcttcagca agttccaaat
     1981 gcacataagg attgggtctg tgccctggga gtggtgccag accacccagt tttgctcagt
     2041 ggctgcagag ggggcatttt gaaagtctgg aacatggata cttttatgcc agtgggagag
     2101 atgaagggtc atgatagtcc tatcaatgcc atatgtgtta attccaccca catttttact
     2161 gcagctgatg atcgaactgt gagaatttgg aaggctggct cgagcggtgg gagctccggt
     2221 gtgagcggct ggcggctgtt caagaagatt agctaagttt aaacggccgc gactctagac
     2281 tgcaggcatg caagctgatt tggctgctgc caccgctgag caaggccgct tcgagcagac
     2341 atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc
     2401 tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa
     2461 caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga gatgtgggag
     2521 gtttttttaa gcaagtaaaa cctctacaaa tgtggtaaaa tcttaattaa gacttctaat
     2581 ataacttcgt atagcataca ttatacgaag ttatggttcc actgttgaca attaatcatc
     2641 ggcatagtat atcggcatag tataatacga caaggtgagg aactaaaccc aggaggcaga
     2701 tcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     2761 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     2821 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     2881 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     2941 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     3001 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     3061 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     3121 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     3181 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
     3241 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     3301 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     3361 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     3421 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     3481 ttgacgagtt cttctgattc gaaatgaccg accaagcgac gcccaaccgg tatcagctca
     3541 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     3601 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     3661 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     3721 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     3781 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     3841 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     3901 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     3961 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     4021 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     4081 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     4141 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     4201 tgtttgcaag cagcagatta cgcgcagaaa aaaaggattt caagaagatc ctttgatctt
     4261 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     4321 attatcaaaa aggatcttca cctagatcct tttatagtcc ggaatcgtca cacaaaaaac
     4381 caacacacag atgtaatgaa aataaagata ttttattgcg gccatcgtga tggctagagt
     4441 ttagccctcc cacacataac cagagggcag caattcacga atcccaactg ccgtcggctg
     4501 tccatcactg tccttcacta tggctttgat cccaggatgc agatcgagaa gcacctgtcg
     4561 gcaccgtccg caggggctca agatgcccct gttctcattt ccgatcgcga cgatacaagt
     4621 caggttgcca gctgccgcag cagcagcagt gcccagcacc acgagttctg cacaaggtcc
     4681 cccagtaaaa tgatatacat tgacaccagt gaagatgcgg ccgtcgctag agagagctgc
     4741 gctggcgacg ctgtagtctt cagagatggg gatgctgttg attgtagccg ttgctctttc
     4801 aatgagggtg gattcttctt gagacaaagg cttggccatg gtggcgctag tgtcagaaga
     4861 atcgagcttt ttgcaaaagc ctaggcctcc aaaaaagcct cctcactact tctggaatag
     4921 ctcagaggcc gaggcggcct cggcctctgc ataaataaaa aaaattagtc agccatgggg
     4981 cggagaatgg gcggaactgg gcggagttag gggcgggatg ggcggagtta ggggcgggac
     5041 tatggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg
     5101 ggactttcca cacctggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc
     5161 tggggagcct ggggactttc cacaccctaa ctgacacaca ttccacagct ggttctttcc
     5221 gcctcagaag gtacctaacc aagttcctct ttcagaggtt atttcaggcc atggtgctgc
     5281 gc
//
