LOCUS       sequence-423353-        6075 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of human COPB2 fragment M1 to A597. N-terminal 
            HiBiT tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6075)
  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 6075)
  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_COPB2:1-597
FEATURES             Location/Qualifiers
     source          1..6075
                     /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    3138..3259
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3149..3168)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(3374..3407)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3417..3464
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3496..4290
                     /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(3550..3569)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4160..4179
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4448..5036)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4528..4547)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5162..5210
                     /note="synthetic polyadenylation signal"
     CDS             complement(5234..5632)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5629..5638
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5663..6020)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5677..5812
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5829..6020
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5977..5997
                     /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 tgcctctgcg
     1261 acttgatatc aaaagaaagc taactgctag atctgatcga gttaagagtg tggatctgca
     1321 tcctacagag ccatggatgt tggcaagtct ttacaatggc agtgtgtgtg tttggaatca
     1381 tgaaacacag acactggtga agacatttga agtatgtgat cttcctgttc gagctgcaaa
     1441 gtttgttgca aggaagaatt gggttgtgac aggagcggat gacatgcaga ttagagtgtt
     1501 caattacaat actctggaga gagttcatat gtttgaagca cactcagact acattcgctg
     1561 tattgctgtt catccaaccc agcctttcat tctaactagc agtgatgaca tgcttattaa
     1621 gctctgggac tgggataaaa aatggtcttg ctcacaagtg tttgaaggac acacccatta
     1681 tgttatgcag attgtgatca accccaaaga taacaatcag tttgccagtg cctctttgga
     1741 caggactatc aaggtgtggc agttgggctc ttcgtcacca aacttcactt tggaaggaca
     1801 tgagaaaggc gtgaattgca ttgattacta cagtggtggg gacaagccat acctcatttc
     1861 aggtgcagat gaccgtcttg ttaaaatatg ggattatcag aataaaacat gtgtgcagac
     1921 actggaagga catgcccaaa atgtgtcttg tgccagcttt catcctgagt tgccaatcat
     1981 tatcacaggt tcagaagatg gaacagtacg tatttggcat tcaagcacct accggcttga
     2041 gagcacactg aattatggaa tggagagggt atggtgcgtg gccagtctaa gagggtcaaa
     2101 caatgtcgct ttgggctatg atgaagggag catcattgtt aagcttggtc gggaggaacc
     2161 tgccatgtcc atggatgcca atggaaagat aatttgggcc aagcattcag aagtccagca
     2221 ggccaaccta aaagcaatgg gagatgctga aattaaagat ggtgaaagat tgccactggc
     2281 agtaaaggat atgggcagtt gtgaaatata ccctcagact attcagcaca atcctaatgg
     2341 gcggtttgtg gtggtgtgtg gtgatgggga gtatatcatc tacacagcaa tggcattgag
     2401 aaacaagagc tttggatctg ctcaggagtt tgcatgggcc cacgattctt cagagtatgc
     2461 aataagagag agcaacagca ttgtaaagat atttaagaac tttaaggaaa aaaaatcatt
     2521 taaaccagat tttggagcag aaagtatcta cggcggcttc ttattgggag tcagatctgt
     2581 aaatggctta gccttctatg actgggacaa tacagaactc atacgaagaa ttgaaattca
     2641 gcccaaacat attttctggt ctgactctgg agagctagtc tgtattgcta ctgaggaatc
     2701 attttttatc cttaagtatc tgtcagaaaa agtcttggct gcacaggaaa cacatgaggg
     2761 agttactgaa gatggcattg aagatgcctt tgaggttctt ggtgagattc aggaaattgt
     2821 gaaaacaggg ctttgggtag gcgattgctt catttacaca agttctgtga acagattaaa
     2881 ttattatgtt ggaggagaaa tagtcaccat tgcccacttg gacaggacga tgtatctcct
     2941 aggctacatt cctaaagaca acaggcttta tctgggggat aaagaattga acatcattag
     3001 ctattccctg ctggtttcag tcctggaata ccagacagct taatctagag gtcaagcttc
     3061 gggggtttaa acctgcaggc atgcaagctg atttggctgc tgccaccgct gagcaaggcc
     3121 gcttcgagca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca
     3181 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
     3241 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg
     3301 ggagatgtgg gaggtttttt taagcaagta aaacctctac aaatgtggta aaatcttaat
     3361 taagacttct aatataactt cgtatagcat acattatacg aagttatggt tccactgttg
     3421 acaattaatc atcggcatag tatatcggca tagtataata cgacaaggtg aggaactaaa
     3481 cccaggaggc agatcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     3541 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     3601 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     3661 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     3721 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     3781 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     3841 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     3901 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     3961 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     4021 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     4081 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     4141 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     4201 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     4261 ttctatcgcc ttcttgacga gttcttctga ttcgaaatga ccgaccaagc gacgcccaac
     4321 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     4381 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     4441 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     4501 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     4561 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     4621 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     4681 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     4741 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     4801 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     4861 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
     4921 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     4981 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tttcaagaag
     5041 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     5101 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttatag tccggaatcg
     5161 tcacacaaaa aaccaacaca cagatgtaat gaaaataaag atattttatt gcggccatcg
     5221 tgatggctag agtttagccc tcccacacat aaccagaggg cagcaattca cgaatcccaa
     5281 ctgccgtcgg ctgtccatca ctgtccttca ctatggcttt gatcccagga tgcagatcga
     5341 gaagcacctg tcggcaccgt ccgcaggggc tcaagatgcc cctgttctca tttccgatcg
     5401 cgacgataca agtcaggttg ccagctgccg cagcagcagc agtgcccagc accacgagtt
     5461 ctgcacaagg tcccccagta aaatgatata cattgacacc agtgaagatg cggccgtcgc
     5521 tagagagagc tgcgctggcg acgctgtagt cttcagagat ggggatgctg ttgattgtag
     5581 ccgttgctct ttcaatgagg gtggattctt cttgagacaa aggcttggcc atggtggcgc
     5641 tagtgtcaga agaatcgagc tttttgcaaa agcctaggcc tccaaaaaag cctcctcact
     5701 acttctggaa tagctcagag gccgaggcgg cctcggcctc tgcataaata aaaaaaatta
     5761 gtcagccatg gggcggagaa tgggcggaac tgggcggagt taggggcggg atgggcggag
     5821 ttaggggcgg gactatggtt gctgactaat tgagatgcat gctttgcata cttctgcctg
     5881 ctggggagcc tggggacttt ccacacctgg ttgctgacta attgagatgc atgctttgca
     5941 tacttctgcc tgctggggag cctggggact ttccacaccc taactgacac acattccaca
     6001 gctggttctt tccgcctcag aaggtaccta accaagttcc tctttcagag gttatttcag
     6061 gccatggtgc tgcgc
//
