LOCUS       sequence-422633-        6119 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of human COPB2 fragment M1 to A597. C-terminal 
            HiBiT tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6119)
  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 6119)
  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_COPB2:1-597
FEATURES             Location/Qualifiers
     source          1..6119
                     /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    3182..3303
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3193..3212)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(3418..3451)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3461..3508
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3540..4334
                     /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(3594..3613)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4204..4223
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4492..5080)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4572..4591)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5206..5254
                     /note="synthetic polyadenylation signal"
     CDS             complement(5278..5676)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5673..5682
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5707..6064)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5721..5856
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5873..6064
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     6021..6041
                     /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 ccatgcctct gcgacttgat
     1261 atcaaaagaa agctaactgc tagatctgat cgagttaaga gtgtggatct gcatcctaca
     1321 gagccatgga tgttggcaag tctttacaat ggcagtgtgt gtgtttggaa tcatgaaaca
     1381 cagacactgg tgaagacatt tgaagtatgt gatcttcctg ttcgagctgc aaagtttgtt
     1441 gcaaggaaga attgggttgt gacaggagcg gatgacatgc agattagagt gttcaattac
     1501 aatactctgg agagagttca tatgtttgaa gcacactcag actacattcg ctgtattgct
     1561 gttcatccaa cccagccttt cattctaact agcagtgatg acatgcttat taagctctgg
     1621 gactgggata aaaaatggtc ttgctcacaa gtgtttgaag gacacaccca ttatgttatg
     1681 cagattgtga tcaaccccaa agataacaat cagtttgcca gtgcctcttt ggacaggact
     1741 atcaaggtgt ggcagttggg ctcttcgtca ccaaacttca ctttggaagg acatgagaaa
     1801 ggcgtgaatt gcattgatta ctacagtggt ggggacaagc catacctcat ttcaggtgca
     1861 gatgaccgtc ttgttaaaat atgggattat cagaataaaa catgtgtgca gacactggaa
     1921 ggacatgccc aaaatgtgtc ttgtgccagc tttcatcctg agttgccaat cattatcaca
     1981 ggttcagaag atggaacagt acgtatttgg cattcaagca cctaccggct tgagagcaca
     2041 ctgaattatg gaatggagag ggtatggtgc gtggccagtc taagagggtc aaacaatgtc
     2101 gctttgggct atgatgaagg gagcatcatt gttaagcttg gtcgggagga acctgccatg
     2161 tccatggatg ccaatggaaa gataatttgg gccaagcatt cagaagtcca gcaggccaac
     2221 ctaaaagcaa tgggagatgc tgaaattaaa gatggtgaaa gattgccact ggcagtaaag
     2281 gatatgggca gttgtgaaat ataccctcag actattcagc acaatcctaa tgggcggttt
     2341 gtggtggtgt gtggtgatgg ggagtatatc atctacacag caatggcatt gagaaacaag
     2401 agctttggat ctgctcagga gtttgcatgg gcccacgatt cttcagagta tgcaataaga
     2461 gagagcaaca gcattgtaaa gatatttaag aactttaagg aaaaaaaatc atttaaacca
     2521 gattttggag cagaaagtat ctacggcggc ttcttattgg gagtcagatc tgtaaatggc
     2581 ttagccttct atgactggga caatacagaa ctcatacgaa gaattgaaat tcagcccaaa
     2641 catattttct ggtctgactc tggagagcta gtctgtattg ctactgagga atcatttttt
     2701 atccttaagt atctgtcaga aaaagtcttg gctgcacagg aaacacatga gggagttact
     2761 gaagatggca ttgaagatgc ctttgaggtt cttggtgaga ttcaggaaat tgtgaaaaca
     2821 gggctttggg taggcgattg cttcatttac acaagttctg tgaacagatt aaattattat
     2881 gttggaggag aaatagtcac cattgcccac ttggacagga cgatgtatct cctaggctac
     2941 attcctaaag acaacaggct ttatctgggg gataaagaat tgaacatcat tagctattcc
     3001 ctgctggttt cagtcctgga ataccagaca gctggctcga gcggtgggag ctccggtgtg
     3061 agcggctggc ggctgttcaa gaagattagc taagtttaaa cggccgcgac tctagactgc
     3121 aggcatgcaa gctgatttgg ctgctgccac cgctgagcaa ggccgcttcg agcagacatg
     3181 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
     3241 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
     3301 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggagat gtgggaggtt
     3361 tttttaagca agtaaaacct ctacaaatgt ggtaaaatct taattaagac ttctaatata
     3421 acttcgtata gcatacatta tacgaagtta tggttccact gttgacaatt aatcatcggc
     3481 atagtatatc ggcatagtat aatacgacaa ggtgaggaac taaacccagg aggcagatca
     3541 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     3601 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     3661 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     3721 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     3781 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     3841 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     3901 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     3961 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     4021 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
     4081 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     4141 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     4201 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     4261 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     4321 acgagttctt ctgattcgaa atgaccgacc aagcgacgcc caaccggtat cagctcactc
     4381 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     4441 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     4501 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     4561 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     4621 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     4681 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     4741 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     4801 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     4861 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     4921 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     4981 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     5041 ttgcaagcag cagattacgc gcagaaaaaa aggatttcaa gaagatcctt tgatcttttc
     5101 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     5161 atcaaaaagg atcttcacct agatcctttt atagtccgga atcgtcacac aaaaaaccaa
     5221 cacacagatg taatgaaaat aaagatattt tattgcggcc atcgtgatgg ctagagttta
     5281 gccctcccac acataaccag agggcagcaa ttcacgaatc ccaactgccg tcggctgtcc
     5341 atcactgtcc ttcactatgg ctttgatccc aggatgcaga tcgagaagca cctgtcggca
     5401 ccgtccgcag gggctcaaga tgcccctgtt ctcatttccg atcgcgacga tacaagtcag
     5461 gttgccagct gccgcagcag cagcagtgcc cagcaccacg agttctgcac aaggtccccc
     5521 agtaaaatga tatacattga caccagtgaa gatgcggccg tcgctagaga gagctgcgct
     5581 ggcgacgctg tagtcttcag agatggggat gctgttgatt gtagccgttg ctctttcaat
     5641 gagggtggat tcttcttgag acaaaggctt ggccatggtg gcgctagtgt cagaagaatc
     5701 gagctttttg caaaagccta ggcctccaaa aaagcctcct cactacttct ggaatagctc
     5761 agaggccgag gcggcctcgg cctctgcata aataaaaaaa attagtcagc catggggcgg
     5821 agaatgggcg gaactgggcg gagttagggg cgggatgggc ggagttaggg gcgggactat
     5881 ggttgctgac taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga
     5941 ctttccacac ctggttgctg actaattgag atgcatgctt tgcatacttc tgcctgctgg
     6001 ggagcctggg gactttccac accctaactg acacacattc cacagctggt tctttccgcc
     6061 tcagaaggta cctaaccaag ttcctctttc agaggttatt tcaggccatg gtgctgcgc
//
