LOCUS       sequence-423304-        7341 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human CORO1A. C-terminal NanoLuc
            luciferase tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7341)
  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 7341)
  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-C_CORO1A:1-461
FEATURES             Location/Qualifiers
     source          1..7341
                     /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             2516..3028
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     primer_bind     complement(3104..3122)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3159..3280
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3170..3189)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3224..3243
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3525..3882
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3525..3716
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3548..3568)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3733..3868
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3913..4950
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             4929..4937
                     /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    4974..5022
                     /note="synthetic polyadenylation signal"
     promoter        5178..5282
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5283..5351,5352..6143)
                     /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(5501..5520)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(6301..6889)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6381..6400)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    7005..7288
                     /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 ggctagcaaa gcgatcgctt ccgaattcgc
     1081 caccatgagc cggcaggtgg tccgctccag caagttccgc cacgtgtttg gacagccggc
     1141 caaggccgac cagtgctatg aagatgtgcg cgtctcacag accacctggg acagtggctt
     1201 ctgtgctgtc aaccctaagt ttgtggccct gatctgtgag gccagcgggg gaggggcctt
     1261 cctggtgctg cccctgggca agactggacg tgtggacaag aatgcgccca cggtctgtgg
     1321 ccacacagcc cctgtgctag acatcgcctg gtgcccgcac aatgacaacg tcattgccag
     1381 tggctccgag gactgcacag tcatggtgtg ggagatcccg gatgggggcc tgatgctgcc
     1441 cctgcgggag cccgtcgtca ccctggaggg ccacaccaag cgtgtgggca ttgtggcctg
     1501 gcacaccaca gcccagaacg tgctgctcag tgcaggttgt gacaacgtga tcatggtgtg
     1561 ggacgtgggc actggggcgg ccatgctgac actgggccca gaggtgcacc cagacacgat
     1621 ctacagtgtg gactggagcc gagatggagg cctcatttgt acctcctgcc gtgacaagcg
     1681 cgtgcgcatc atcgagcccc gcaaaggcac tgtcgtagct gagaaggacc gtccccacga
     1741 ggggacccgg cccgtgcgtg cagtgttcgt gtcggagggg aagatcctga ccacgggctt
     1801 cagccgcatg agtgagcggc aggtggcgct gtgggacaca aagcacctgg aggagccgct
     1861 gtccctgcag gagctggaca ccagcagcgg tgtcctgctg cccttctttg accctgacac
     1921 caacatcgtc tacctctgtg gcaagggtga cagctcaatc cggtactttg agatcacttc
     1981 cgaggcccct ttcctgcact atctctccat gttcagttcc aaggagtccc agcggggcat
     2041 gggctacatg cccaaacgtg gcctggaggt gaacaagtgt gagatcgcca ggttctacaa
     2101 gctgcacgag cggaggtgtg agcccattgc catgacagtg cctcgaaagt cggacctgtt
     2161 ccaggaggac ctgtacccac ccaccgcagg gcccgaccct gccctcacgg ctgaggagtg
     2221 gctggggggt cgggatgctg ggcccctcct catctccctc aaggatggct acgtaccccc
     2281 aaagagccgg gagctgaggg tcaaccgggg cctggacacc gggcgcagga gggcagcacc
     2341 agaggccagt ggcactccca gctcggatgc cgtgtctcgg ctggaggagg agatgcggaa
     2401 gctccaggcc acggtgcagg agctccagaa gcgcttggac aggctggagg agacagtcca
     2461 ggccaagtct agatttgggc ccaattcctg caggcgagct ctcggctcga gcggcgtctt
     2521 cacactcgaa gatttcgttg gggactggcg acagacagcc ggctacaacc tggaccaagt
     2581 ccttgaacag ggaggtgtgt ccagtttgtt tcagaatctc ggggtgtccg taactccgat
     2641 ccaaaggatt gtcctgagcg gtgaaaatgg gctgaagatc gacatccatg tcatcatccc
     2701 gtatgaaggt ctgagcggcg accaaatggg ccagatcgaa aaaattttta aggtggtgta
     2761 ccctgtggat gatcatcact ttaaggtgat cctgcactat ggcacactgg taatcgacgg
     2821 ggttacgccg aacatgatcg actatttcgg acggccgtat gaaggcatcg ccgtgttcga
     2881 cggcaaaaag atcactgtaa cagggaccct gtggaacggc aacaaaatta tcgacgagcg
     2941 cctgatcaac cccgacggct ccctgctgtt ccgagtaacc atcaacggag tgaccggctg
     3001 gcggctgtgc gaacgcattc tggcgtaagg ccgcgactct agggcatgca agctgatccg
     3061 gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga gcaataacta
     3121 gcataacccc ttggggcggc cgcttcgagc agacatgata agatacattg atgagtttgg
     3181 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     3241 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     3301 ttttatgttt caggttcagg gggagatgtg ggaggttttt ttaagcaagt aaaacctcta
     3361 caaatgtggt aaaatcgaat tttaacaaaa tattaacgct tacaatttcc tgatgcggta
     3421 ttttctcctt acgcatctgt gcggtatttc acaccgcata cgcggatctg cgcagcacca
     3481 tggcctgaaa taacctctga aagaggaact tggttaggta ccttctgagg cggaaagaac
     3541 cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     3601 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     3661 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     3721 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     3781 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     3841 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctcga ttcttctgac
     3901 actagcgcca ccatgaagaa gcccgaactc accgctacca gcgttgaaaa atttctcatc
     3961 gagaagttcg acagtgtgag cgacctgatg cagttgtcgg agggcgaaga gagccgagcc
     4021 ttcagcttcg atgtcggcgg acgcggctat gtactgcggg tgaatagctg cgctgatggc
     4081 ttctacaaag accgctacgt gtaccgccac ttcgccagcg ctgcactacc catccccgaa
     4141 gtgttggaca tcggcgagtt cagcgagagc ctgacatact gcatcagtag acgcgcccaa
     4201 ggcgttactc tccaagacct ccccgaaaca gagctgcctg ctgtgttaca gcctgtcgcc
     4261 gaagctatgg atgctattgc cgccgccgac ctcagtcaaa ccagcggctt cggcccattc
     4321 gggccccaag gcatcggcca gtacacaacc tggcgggatt tcatttgcgc cattgctgat
     4381 ccccatgtct accactggca gaccgtgatg gacgacaccg tgtccgccag cgtagctcaa
     4441 gccctggacg aactgatgct gtgggccgaa gactgtcccg aggtgcgcca cctcgtccat
     4501 gccgacttcg gcagcaacaa cgtcctgacc gacaacggcc gcatcaccgc cgtaatcgac
     4561 tggtccgaag ctatgttcgg ggacagtcag tacgaggtgg ccaacatctt cttctggcgg
     4621 ccctggctgg cttgcatgga gcagcagact cgctacttcg agcgccggca tcccgagctg
     4681 gccggcagcc ctcgtctgcg agcctacatg ctgcgcatcg gcctggatca gctctaccag
     4741 agcctcgtgg acggcaactt cgacgatgct gcctgggctc aaggccgctg cgatgccatc
     4801 gtccgcagcg gggccggcac cgtcggtcgc acacaaatcg ctcgccggag cgcagccgta
     4861 tggaccgacg gctgcgtcga ggtgctggcc gacagcggca accgccggcc cagtacacga
     4921 ccgcgcgcta aggaggtagg tcgagtttag actctagcca tcacgatggc cgcaataaaa
     4981 tatctttatt ttcattacat ctgtgtgttg gttttttgtg tgaatcgata gcgataagga
     5041 tcctctttgc gcttgcgttt tcccttgtcc agatagccca gtagctgaca ttcatccggg
     5101 gtcagcaccg tttctgcgga ctggctttct acgtaatggt ttcttagacg tcaggtggca
     5161 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     5221 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     5281 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     5341 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     5401 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     5461 ccgaagaacg ttttccaatg atgagcactt tcaaagttct gctatgtggc gcggtattat
     5521 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     5581 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     5641 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacta
     5701 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     5761 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     5821 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     5881 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     5941 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     6001 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     6061 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     6121 cctcactgat taagcattgg taattcgaaa tgaccgacca agcgacgccc aaccggtatc
     6181 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     6241 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     6301 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     6361 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     6421 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     6481 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     6541 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     6601 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     6661 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     6721 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     6781 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     6841 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatttcaag aagatccttt
     6901 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     6961 catgagatta tcaaaaagga tcttcaccta gatcctttta tagtccggaa atacaggaac
     7021 gcacgctgga tggcccttcg ctgggatggt gaaaccatga aaaatggcag cttcagtgga
     7081 ttaagtgggg gtaatgtggc ctgtaccctc tggttgcata ggtattcata cggttaaaat
     7141 ttatcaggcg cgattgcggc agtttttcgg gtggtttgtt gccattttta cctgtctgct
     7201 gccgtgatcg cgctgaacgc gttttagcgg tgcgtacaat taagggatta tggtaaatcc
     7261 acttactgtc tgccctcgta gccatcgaga taaaccgcag tactccggcc acgatgcgtc
     7321 cggcgtagag gatcgagatc t
//
