LOCUS       sequence-423327-        8208 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human SEC23B. N-terminal NanoLuc
            luciferase tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8208)
  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 8208)
  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: pNLF1-N_SEC23B:1-767
FEATURES             Location/Qualifiers
     source          1..8208
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..721
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     672..692
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          857..989
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     972..991
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1033..1052
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1033..1051
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1065..1577
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     primer_bind     complement(3976..3994)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    4026..4147
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4037..4056)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4091..4110
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4392..4749
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4392..4583
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4415..4435)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4600..4735
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4780..5817
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             5796..5804
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     polyA_signal    5841..5889
                     /note="synthetic polyadenylation signal"
     promoter        6045..6149
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6150..6218,6219..7010)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6368..6387)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(7168..7756)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7248..7267)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    7872..8155
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      661 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      721 tggtttagtg aaccgtcaga tcactagaag ctttattgcg gtagtttatc acagttaaat
      781 tgctaacgca gtcagtgctt ctgacacaac agtctcgaac ttaagctgca gaagttggtc
      841 gtgaggcact gggcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa
      901 actgggcttg tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac
      961 tgacatccac tttgcctttc tctccacagg tgtccactcc cagttcaatt acagctctta
     1021 aggctagagt attaatacga ctcactatag ggctagcgct caccatggtc ttcacactcg
     1081 aagatttcgt tggggactgg cgacagacag ccggctacaa cctggaccaa gtccttgaac
     1141 agggaggtgt gtccagtttg tttcagaatc tcggggtgtc cgtaactccg atccaaagga
     1201 ttgtcctgag cggtgaaaat gggctgaaga tcgacatcca tgtcatcatc ccgtatgaag
     1261 gtctgagcgg cgaccaaatg ggccagatcg aaaaaatttt taaggtggtg taccctgtgg
     1321 atgatcatca ctttaaggtg atcctgcact atggcacact ggtaatcgac ggggttacgc
     1381 cgaacatgat cgactatttc ggacggccgt atgaaggcat cgccgtgttc gacggcaaaa
     1441 agatcactgt aacagggacc ctgtggaacg gcaacaaaat tatcgacgag cgcctgatca
     1501 accccgacgg ctccctgctg ttccgagtaa ccatcaacgg agtgaccggc tggcggctgt
     1561 gcgaacgcat tctggcgggc tcgagcggcg cgatcgcttc cgaattcatg gcgacatacc
     1621 tggagttcat ccagcagaat gaagaacggg atggtgtgcg ttttagttgg aacgtgtggc
     1681 cttccagccg gctggaggct acaagaatgg ttgtacccct ggcttgtctc cttactcctt
     1741 tgaaagaacg tccagaccta cctcctgtac aatatgaacc tgtgctttgc agcaggccaa
     1801 cttgtaaagc tgttctcaac ccactttgtc aggttgatta tcgagcaaaa ctttgggcct
     1861 gtaatttctg ttttcaaaga aatcagtttc ctccagctta tggaggcata tctgaggtga
     1921 atcaacctgc cgaattgatg ccccagtttt ctacaattga gtacgtgata cagcgaggtg
     1981 ctcagtcccc tctgatcttt ctctatgtgg ttgacacatg cctggaggaa gatgaccttc
     2041 aagcactcaa agagtccctg cagatgtccc tgagtcttct tcctccagat gctctggtgg
     2101 gtctgatcac atttggaagg atggtgcagg ttcatgagct aagctgtgaa ggaatctcca
     2161 aaagttatgt cttccgaggg accaaggatt taactgcaaa gcaaatacag gatatgttgg
     2221 gcctgaccaa gccagccatg cccatgcagc aagcacgacc tgcacaacca caggagcacc
     2281 cttttgcttc aagcagattt ctgcagcctg ttcacaagat tgatatgaac ctcactgatc
     2341 ttcttgggga gctacagagg gacccatggc cagtaactca ggggaagaga cctttgcgat
     2401 ccactggtgt ggctttgtcc attgctgttg gcttgctgga gggcactttt ccaaacacag
     2461 gagccaggat catgctgttt actggaggtc cccctaccca agggcctggc atggtggttg
     2521 gagatgaatt aaagattcct attcgttctt ggcatgatat tgagaaagat aatgcacgat
     2581 tcatgaaaaa ggcaaccaag cactatgaga tgcttgctaa tcgaacagct gcaaatggtc
     2641 actgcattga tatttatgct tgtgcccttg atcaaactgg acttttggag atgaagtgtt
     2701 gtgcaaatct tactggaggc tacatggtaa tgggagattc tttcaacact tctctcttca
     2761 agcagacatt ccaaagaatc tttactaaag attttaatgg agatttccga atggcatttg
     2821 gtgctacttt ggacgtaaag acctctcggg aactgaagat tgcaggagcc attggtccat
     2881 gcgtatctct gaatgtgaaa ggaccgtgtg tgtcagaaaa tgagcttggt gttggtggca
     2941 cgagtcagtg gaaaatctgt ggcctagatc ctacatctac acttggcatc tattttgaag
     3001 ttgtcaatca gcacaacacc ccgatccccc aaggaggcag aggagccatc cagtttgtca
     3061 cgcattatca gcactccagc acccagagac gcatccgcgt gaccaccatc gcccgaaatt
     3121 gggcagatgt acagagtcag ctcaggcaca tagaagcagc atttgaccag gaggctgcgg
     3181 cagtgttgat ggcacggctt ggggtgttcc gagcggagtc agaggagggg cccgatgtgc
     3241 tccggtggct ggaccgacaa ctcatccgac tgtgtcaaaa gtttggacag tataacaaag
     3301 aagaccccac ttcttttagg ttatcagatt ccttttctct atatcctcag tttatgttcc
     3361 atctgagaag atctccattt cttcaagtgt ttaacaacag tcctgatgag tcgtcatatt
     3421 acagacatca ttttgcccgg caggacctga cccagtccct catcatgatc cagcccattc
     3481 tctactctta ctcctttcat gggccaccag agccagtact cttggatagc agcagcattc
     3541 tagctgacag aattttgctg atggatactt tctttcaaat tgtcatttat cttggtgaga
     3601 ccatagccca gtggcgtaaa gctggctacc aggacatgcc cgagtatgaa aacttcaagc
     3661 accttctgca ggcaccactg gatgatgctc aagaaattct gcaagcacgc ttcccgatgc
     3721 cacgttacat caacacggag catggaggca gtcaggctcg attccttttg tccaaagtga
     3781 acccatctca gacacacaat aacctgtatg cttggggaca ggaaactgga gcacccatcc
     3841 taactgatga tgttagcctg caggtgttca tggaccattt gaagaagctg gctgtctcca
     3901 gtgcctgtta ataatctaga tttgggccca attcctgcag gattttgcgg ccgcttgctg
     3961 agttggctgc tgccaccgct gagcaataac tagcataacc ccttggccgc ttcgagcaga
     4021 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     4081 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     4141 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg agatgtggga
     4201 ggttttttta agcaagtaaa acctctacaa atgtggtaaa atcgaatttt aacaaaatat
     4261 taacgcttac aatttcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     4321 ccgcatacgc ggatctgcgc agcaccatgg cctgaaataa cctctgaaag aggaacttgg
     4381 ttaggtacct tctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga
     4441 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     4501 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     4561 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     4621 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     4681 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     4741 ttttgcaaaa agctcgattc ttctgacact agcgccacca tgaagaagcc cgaactcacc
     4801 gctaccagcg ttgaaaaatt tctcatcgag aagttcgaca gtgtgagcga cctgatgcag
     4861 ttgtcggagg gcgaagagag ccgagccttc agcttcgatg tcggcggacg cggctatgta
     4921 ctgcgggtga atagctgcgc tgatggcttc tacaaagacc gctacgtgta ccgccacttc
     4981 gccagcgctg cactacccat ccccgaagtg ttggacatcg gcgagttcag cgagagcctg
     5041 acatactgca tcagtagacg cgcccaaggc gttactctcc aagacctccc cgaaacagag
     5101 ctgcctgctg tgttacagcc tgtcgccgaa gctatggatg ctattgccgc cgccgacctc
     5161 agtcaaacca gcggcttcgg cccattcggg ccccaaggca tcggccagta cacaacctgg
     5221 cgggatttca tttgcgccat tgctgatccc catgtctacc actggcagac cgtgatggac
     5281 gacaccgtgt ccgccagcgt agctcaagcc ctggacgaac tgatgctgtg ggccgaagac
     5341 tgtcccgagg tgcgccacct cgtccatgcc gacttcggca gcaacaacgt cctgaccgac
     5401 aacggccgca tcaccgccgt aatcgactgg tccgaagcta tgttcgggga cagtcagtac
     5461 gaggtggcca acatcttctt ctggcggccc tggctggctt gcatggagca gcagactcgc
     5521 tacttcgagc gccggcatcc cgagctggcc ggcagccctc gtctgcgagc ctacatgctg
     5581 cgcatcggcc tggatcagct ctaccagagc ctcgtggacg gcaacttcga cgatgctgcc
     5641 tgggctcaag gccgctgcga tgccatcgtc cgcagcgggg ccggcaccgt cggtcgcaca
     5701 caaatcgctc gccggagcgc agccgtatgg accgacggct gcgtcgaggt gctggccgac
     5761 agcggcaacc gccggcccag tacacgaccg cgcgctaagg aggtaggtcg agtttagact
     5821 ctagccatca cgatggccgc aataaaatat ctttattttc attacatctg tgtgttggtt
     5881 ttttgtgtga atcgatagcg ataaggatcc tctttgcgct tgcgttttcc cttgtccaga
     5941 tagcccagta gctgacattc atccggggtc agcaccgttt ctgcggactg gctttctacg
     6001 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     6061 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     6121 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     6181 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     6241 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     6301 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttca
     6361 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     6421 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     6481 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     6541 ctgcggccaa cttacttctg acaactatcg gaggaccgaa ggagctaacc gcttttttgc
     6601 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     6661 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     6721 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     6781 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     6841 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     6901 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     6961 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ttcgaaatga
     7021 ccgaccaagc gacgcccaac cggtatcagc tcactcaaag gcggtaatac ggttatccac
     7081 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     7141 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     7201 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     7261 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     7321 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     7381 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     7441 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     7501 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     7561 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     7621 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     7681 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     7741 aaaaaaagga tttcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     7801 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     7861 ccttttatag tccggaaata caggaacgca cgctggatgg cccttcgctg ggatggtgaa
     7921 accatgaaaa atggcagctt cagtggatta agtgggggta atgtggcctg taccctctgg
     7981 ttgcataggt attcatacgg ttaaaattta tcaggcgcga ttgcggcagt ttttcgggtg
     8041 gtttgttgcc atttttacct gtctgctgcc gtgatcgcgc tgaacgcgtt ttagcggtgc
     8101 gtacaattaa gggattatgg taaatccact tactgtctgc cctcgtagcc atcgagataa
     8161 accgcagtac tccggccacg atgcgtccgg cgtagaggat cgagatct
//
