LOCUS       sequence-425899-        6381 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human DNAI1. N-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6381)
  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 6381)
  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_DNAI1:1-699
FEATURES             Location/Qualifiers
     source          1..6381
                     /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    3444..3565
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3455..3474)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(3680..3713)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3723..3770
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3802..4596
                     /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(3856..3875)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4466..4485
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4754..5342)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4834..4853)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5468..5516
                     /note="synthetic polyadenylation signal"
     CDS             complement(5540..5938)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5935..5944
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5969..6326)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5983..6118
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        6135..6326
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     6283..6303
                     /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 tgattcctgc
     1261 ttctgcgaag tctccccata aacagcctca taagcagagc atcagcatag gcagaggaac
     1321 caggaagaga gatgaagatt cagggactga agtgggagaa ggcacagatg aatgggccca
     1381 atccaaagcc acagttagac cccctgacca gctggagttg accgatgcgg agttaaagga
     1441 ggagttcact cggattttga cagccaacaa cccacacgca ccccagaaca ttgtcaggta
     1501 cagcttcaaa gaaggcacat ataagcctat tggctttgtg aaccaactgg cagttcacta
     1561 cacccaggtt gggaacctga tccccaaaga ctcagatgaa ggacggcggc agcattaccg
     1621 cgatgaatta gtggcaggtt ctcaggagtc tgtcaaggtg atttcagaaa caggaaacct
     1681 cgaagaagac gaagagccca aggagttaga aactgagcct gggagtcaaa cagatgtgcc
     1741 tgcagctggg gcagctgaaa aagtgactga agaagaattg atgactccta agcagcccaa
     1801 ggagagaaag ctcactaacc agttcaactt cagtgagagg gcctcacaga cctgcaacaa
     1861 ccctgtccgg gatcgagaat gccagacgga gcctcctccc aggacaaact tttcagccac
     1921 agccaatcag tgggagatct atgatgccta tgtagaggaa cttgagaagc aggaaaagac
     1981 caaagagaag gagaaggcaa agaccccagt ggctaaaaaa tcagggaaga tggccatgag
     2041 gaagctgaca tctatggagt ctcagactga tgatctcatc aaattgtccc aagctgctaa
     2101 gatcatggag cggatggtca accagaatac atatgatgac attgctcaag attttaagta
     2161 ctatgacgat gctgctgatg aataccggga ccaggtgggt accctgctgc cgctctggaa
     2221 gttccaaaat gacaaagcca agcgcctgtc cgtcactgcc ctctgctgga atccaaagta
     2281 cagggatctg tttgcagtgg gatatggctc ttatgacttc atgaagcaga gccggggcat
     2341 gctgctgctc tacagcctga agaaccccag cttccctgag tacatgttca gcagcaacag
     2401 cggcgtcatg tgtctcgaca tccacgtgga ccacccctac ctggtggcag taggccacta
     2461 tgacggcaac gtggccattt acaacctcaa gaagccccac tcccagccct ccttctgcag
     2521 ctcagccaag tctggcaagc actcagaccc tgtgtggcag gtcaagtggc agaaggatga
     2581 catggaccaa aaccttaact tcttctctgt gtcatctgac ggcaggattg tgtcttggac
     2641 tctcgtgaag agaaagctgg ttcacataga tgtcatcaag ctgaaggtgg aaggcagcac
     2701 cacggaagtt cctgaggggt tgcagctgca ccaagtgggt tgtggcactg cctttgactt
     2761 ccacaaagag attgactaca tgttcctagt gggcacagag gagggaaaaa tctacaagtg
     2821 ctctaaatcc tactccagcc aattcctcga cacctatgac gcccacaaca tgtcagtgga
     2881 cactgtgtcc tggaacccat accacaccaa ggtcttcatg tcctgcagct ccgactggac
     2941 agtgaagatc tgggaccaca ccatcaagac cccgatgttc atctatgacc tgaactcagc
     3001 cgtgggtgat gtggcctggg cgccatactc ttctactgtg ttcgcagcag tcaccacaga
     3061 tgggaaggcc cacatatttg acttagccat caacaagtat gaggccatct gcaaccagcc
     3121 tgtggcggcc aaaaagaaca ggctcaccca cgtgcagttc aatctcatcc accccatcat
     3181 cattgtgggc gatgaccgtg ggcacatcat cagcctcaag ctctcaccca atttgcgcaa
     3241 gatgccaaag gaaaagaagg ggcaggaggt gcagaagggt ccagctgtgg agattgcgaa
     3301 actggacaaa ctgctgaacc tggtgaggga agtgaaaatc aagacctaat ctagaggtca
     3361 agcttcgggg gtttaaacct gcaggcatgc aagctgattt ggctgctgcc accgctgagc
     3421 aaggccgctt cgagcagaca tgataagata cattgatgag tttggacaaa ccacaactag
     3481 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     3541 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     3601 tcagggggag atgtgggagg tttttttaag caagtaaaac ctctacaaat gtggtaaaat
     3661 cttaattaag acttctaata taacttcgta tagcatacat tatacgaagt tatggttcca
     3721 ctgttgacaa ttaatcatcg gcatagtata tcggcatagt ataatacgac aaggtgagga
     3781 actaaaccca ggaggcagat catgattgaa caagatggat tgcacgcagg ttctccggcc
     3841 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     3901 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     3961 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     4021 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     4081 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     4141 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     4201 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     4261 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     4321 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     4381 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     4441 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     4501 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     4561 atcgccttct atcgccttct tgacgagttc ttctgattcg aaatgaccga ccaagcgacg
     4621 cccaaccggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     4681 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     4741 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     4801 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     4861 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     4921 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     4981 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     5041 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     5101 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     5161 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     5221 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     5281 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatttc
     5341 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     5401 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttatagtccg
     5461 gaatcgtcac acaaaaaacc aacacacaga tgtaatgaaa ataaagatat tttattgcgg
     5521 ccatcgtgat ggctagagtt tagccctccc acacataacc agagggcagc aattcacgaa
     5581 tcccaactgc cgtcggctgt ccatcactgt ccttcactat ggctttgatc ccaggatgca
     5641 gatcgagaag cacctgtcgg caccgtccgc aggggctcaa gatgcccctg ttctcatttc
     5701 cgatcgcgac gatacaagtc aggttgccag ctgccgcagc agcagcagtg cccagcacca
     5761 cgagttctgc acaaggtccc ccagtaaaat gatatacatt gacaccagtg aagatgcggc
     5821 cgtcgctaga gagagctgcg ctggcgacgc tgtagtcttc agagatgggg atgctgttga
     5881 ttgtagccgt tgctctttca atgagggtgg attcttcttg agacaaaggc ttggccatgg
     5941 tggcgctagt gtcagaagaa tcgagctttt tgcaaaagcc taggcctcca aaaaagcctc
     6001 ctcactactt ctggaatagc tcagaggccg aggcggcctc ggcctctgca taaataaaaa
     6061 aaattagtca gccatggggc ggagaatggg cggaactggg cggagttagg ggcgggatgg
     6121 gcggagttag gggcgggact atggttgctg actaattgag atgcatgctt tgcatacttc
     6181 tgcctgctgg ggagcctggg gactttccac acctggttgc tgactaattg agatgcatgc
     6241 tttgcatact tctgcctgct ggggagcctg gggactttcc acaccctaac tgacacacat
     6301 tccacagctg gttctttccg cctcagaagg tacctaacca agttcctctt tcagaggtta
     6361 tttcaggcca tggtgctgcg c
//
