LOCUS       sequence-423423-        6425 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human DNAI1. C-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6425)
  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 6425)
  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_DNAI1:1-699
FEATURES             Location/Qualifiers
     source          1..6425
                     /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    3488..3609
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3499..3518)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(3724..3757)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3767..3814
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3846..4640
                     /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(3900..3919)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4510..4529
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4798..5386)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4878..4897)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5512..5560
                     /note="synthetic polyadenylation signal"
     CDS             complement(5584..5982)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5979..5988
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(6013..6370)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6027..6162
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        6179..6370
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     6327..6347
                     /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 ccatgattcc tgcttctgcg
     1261 aagtctcccc ataaacagcc tcataagcag agcatcagca taggcagagg aaccaggaag
     1321 agagatgaag attcagggac tgaagtggga gaaggcacag atgaatgggc ccaatccaaa
     1381 gccacagtta gaccccctga ccagctggag ttgaccgatg cggagttaaa ggaggagttc
     1441 actcggattt tgacagccaa caacccacac gcaccccaga acattgtcag gtacagcttc
     1501 aaagaaggca catataagcc tattggcttt gtgaaccaac tggcagttca ctacacccag
     1561 gttgggaacc tgatccccaa agactcagat gaaggacggc ggcagcatta ccgcgatgaa
     1621 ttagtggcag gttctcagga gtctgtcaag gtgatttcag aaacaggaaa cctcgaagaa
     1681 gacgaagagc ccaaggagtt agaaactgag cctgggagtc aaacagatgt gcctgcagct
     1741 ggggcagctg aaaaagtgac tgaagaagaa ttgatgactc ctaagcagcc caaggagaga
     1801 aagctcacta accagttcaa cttcagtgag agggcctcac agacctgcaa caaccctgtc
     1861 cgggatcgag aatgccagac ggagcctcct cccaggacaa acttttcagc cacagccaat
     1921 cagtgggaga tctatgatgc ctatgtagag gaacttgaga agcaggaaaa gaccaaagag
     1981 aaggagaagg caaagacccc agtggctaaa aaatcaggga agatggccat gaggaagctg
     2041 acatctatgg agtctcagac tgatgatctc atcaaattgt cccaagctgc taagatcatg
     2101 gagcggatgg tcaaccagaa tacatatgat gacattgctc aagattttaa gtactatgac
     2161 gatgctgctg atgaataccg ggaccaggtg ggtaccctgc tgccgctctg gaagttccaa
     2221 aatgacaaag ccaagcgcct gtccgtcact gccctctgct ggaatccaaa gtacagggat
     2281 ctgtttgcag tgggatatgg ctcttatgac ttcatgaagc agagccgggg catgctgctg
     2341 ctctacagcc tgaagaaccc cagcttccct gagtacatgt tcagcagcaa cagcggcgtc
     2401 atgtgtctcg acatccacgt ggaccacccc tacctggtgg cagtaggcca ctatgacggc
     2461 aacgtggcca tttacaacct caagaagccc cactcccagc cctccttctg cagctcagcc
     2521 aagtctggca agcactcaga ccctgtgtgg caggtcaagt ggcagaagga tgacatggac
     2581 caaaacctta acttcttctc tgtgtcatct gacggcagga ttgtgtcttg gactctcgtg
     2641 aagagaaagc tggttcacat agatgtcatc aagctgaagg tggaaggcag caccacggaa
     2701 gttcctgagg ggttgcagct gcaccaagtg ggttgtggca ctgcctttga cttccacaaa
     2761 gagattgact acatgttcct agtgggcaca gaggagggaa aaatctacaa gtgctctaaa
     2821 tcctactcca gccaattcct cgacacctat gacgcccaca acatgtcagt ggacactgtg
     2881 tcctggaacc cataccacac caaggtcttc atgtcctgca gctccgactg gacagtgaag
     2941 atctgggacc acaccatcaa gaccccgatg ttcatctatg acctgaactc agccgtgggt
     3001 gatgtggcct gggcgccata ctcttctact gtgttcgcag cagtcaccac agatgggaag
     3061 gcccacatat ttgacttagc catcaacaag tatgaggcca tctgcaacca gcctgtggcg
     3121 gccaaaaaga acaggctcac ccacgtgcag ttcaatctca tccaccccat catcattgtg
     3181 ggcgatgacc gtgggcacat catcagcctc aagctctcac ccaatttgcg caagatgcca
     3241 aaggaaaaga aggggcagga ggtgcagaag ggtccagctg tggagattgc gaaactggac
     3301 aaactgctga acctggtgag ggaagtgaaa atcaagaccg gctcgagcgg tgggagctcc
     3361 ggtgtgagcg gctggcggct gttcaagaag attagctaag tttaaacggc cgcgactcta
     3421 gactgcaggc atgcaagctg atttggctgc tgccaccgct gagcaaggcc gcttcgagca
     3481 gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa
     3541 tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat
     3601 aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggagatgtgg
     3661 gaggtttttt taagcaagta aaacctctac aaatgtggta aaatcttaat taagacttct
     3721 aatataactt cgtatagcat acattatacg aagttatggt tccactgttg acaattaatc
     3781 atcggcatag tatatcggca tagtataata cgacaaggtg aggaactaaa cccaggaggc
     3841 agatcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     3901 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     3961 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     4021 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     4081 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     4141 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     4201 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     4261 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     4321 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc
     4381 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     4441 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     4501 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     4561 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     4621 ttcttgacga gttcttctga ttcgaaatga ccgaccaagc gacgcccaac cggtatcagc
     4681 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     4741 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     4801 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     4861 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     4921 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     4981 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     5041 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     5101 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     5161 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     5221 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     5281 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     5341 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tttcaagaag atcctttgat
     5401 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     5461 gagattatca aaaaggatct tcacctagat ccttttatag tccggaatcg tcacacaaaa
     5521 aaccaacaca cagatgtaat gaaaataaag atattttatt gcggccatcg tgatggctag
     5581 agtttagccc tcccacacat aaccagaggg cagcaattca cgaatcccaa ctgccgtcgg
     5641 ctgtccatca ctgtccttca ctatggcttt gatcccagga tgcagatcga gaagcacctg
     5701 tcggcaccgt ccgcaggggc tcaagatgcc cctgttctca tttccgatcg cgacgataca
     5761 agtcaggttg ccagctgccg cagcagcagc agtgcccagc accacgagtt ctgcacaagg
     5821 tcccccagta aaatgatata cattgacacc agtgaagatg cggccgtcgc tagagagagc
     5881 tgcgctggcg acgctgtagt cttcagagat ggggatgctg ttgattgtag ccgttgctct
     5941 ttcaatgagg gtggattctt cttgagacaa aggcttggcc atggtggcgc tagtgtcaga
     6001 agaatcgagc tttttgcaaa agcctaggcc tccaaaaaag cctcctcact acttctggaa
     6061 tagctcagag gccgaggcgg cctcggcctc tgcataaata aaaaaaatta gtcagccatg
     6121 gggcggagaa tgggcggaac tgggcggagt taggggcggg atgggcggag ttaggggcgg
     6181 gactatggtt gctgactaat tgagatgcat gctttgcata cttctgcctg ctggggagcc
     6241 tggggacttt ccacacctgg ttgctgacta attgagatgc atgctttgca tacttctgcc
     6301 tgctggggag cctggggact ttccacaccc taactgacac acattccaca gctggttctt
     6361 tccgcctcag aaggtaccta accaagttcc tctttcagag gttatttcag gccatggtgc
     6421 tgcgc
//
