LOCUS       sequence-379950-        8472 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the Nanoluciferase fused to the human parathyroid hormone 
            receptor type 1 that is missing the extracellular domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8472)
  AUTHORS   Yu Z, Cary BP, Kim TW, Nguyen KD, Gardella TJ, Gellman SH
  TITLE     Kinetic and Thermodynamic Insights into Agonist Interactions with 
            the Parathyroid Hormone Receptor-1 from a New NanoBRET Assay.
  JOURNAL   ACS Chem Biol. 2022 Nov 18;17(11):3148-3158. doi: 
            10.1021/acschembio.2c00595. Epub 2022 Oct 25.
  PUBMED    36282520
REFERENCE   2  (bases 1 to 8472)
  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/acschembio.2c00595"; journalName: "ACS Chem Biol"; date: 
            "2022-11-18- 18"; volume: "17"; issue: "11"; pages: "3148-3158"
COMMENT     Sequence Label: nLuc-Delta-ECD-PTHR1
FEATURES             Location/Qualifiers
     source          1..8472
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        49..352
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        354..556
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     507..527
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             630..656
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             726..1235
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     polyA_signal    2631..2752
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2642..2661)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2696..2715
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2867..2913
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2943..3079
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3124..3195
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(3421..4009)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3501..3520)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        4509..5718
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     5555..5574
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             5722..6750
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    6777..6825
                     /note="synthetic polyadenylation signal"
     CDS             join(7091..7159,7160..7951)
                     /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(7309..7328)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      7964..8100
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    8252..8323
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(8328..8374)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    8392..8440
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
       61 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
      121 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
      181 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
      241 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
      301 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tgctgatgcg
      361 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
      421 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
      481 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
      541 ctatataagc agagctggtt tagtgaaccg tcagatcaga tctttgtcga tcctaccatc
      601 cactcgacac acccgccagc ggccgcatgt acccatacga tgttccagat tacgctgatg
      661 ggaccgcccg gatcgcaccc ggcctggcgc tcctgctctg ctgccccgtg ctcagctccg
      721 cgtacgtctt cacactcgaa gatttcgttg gggactggcg acagacagcc ggctacaacc
      781 tggaccaagt ccttgaacag ggaggtgtgt ccagtttgtt tcagaatctc ggggtgtccg
      841 taactccgat ccaaaggatt gtcctgagcg gtgaaaatgg gctgaagatc gacatccatg
      901 tcatcatccc gtatgaaggt ctgagcggcg accaaatggg ccagatcgaa aaaattttta
      961 aggtggtgta ccctgtggat gatcatcact ttaaggtgat cctgcactat ggcacactgg
     1021 taatcgacgg ggttacgccg aacatgatcg actatttcgg acggccgtat gaaggcatcg
     1081 ccgtgttcga cggcaaaaag atcactgtaa cagggaccct gtggaacggc aacaaaatta
     1141 tcgacgagcg cctgatcaac cccgacggct ccctgctgtt ccgagtaacc atcaacggag
     1201 tgaccggctg gcggctgtgc gaacgcattc tggcgggtgg aggtggatca tttctcacca
     1261 atgagactcg tgaacgggag gtgtttgacc gcctgggcat gatttacacc gtgggctact
     1321 ccgtgtccct ggcgtccctc accgtagctg tgctcatcct ggcctacttt aggcggctgc
     1381 actgcacgcg caactacatc cacatgcacc tgttcctgtc cttcatgctg cgcgccgtga
     1441 gcatcttcgt caaggacgct gtgctctact ctggcgccac gcttgatgag gctgagcgcc
     1501 tcaccgagga ggagctgcgc gccatcgccc aggcgccccc gccgcctgcc accgccgctg
     1561 ccggctacgc gggctgcagg gtggctgtga ccttcttcct ttacttcctg gccaccaact
     1621 actactggat tctggtggag gggctgtacc tgcacagcct catcttcatg gccttcttct
     1681 cagagaagaa gtacctgtgg ggcttcacag tcttcggctg gggtctgccc gctgtcttcg
     1741 tggctgtgtg ggtcagtgtc agagctaccc tggccaacac cgggtgctgg gacttgagct
     1801 ccgggaacaa aaagtggatc atccaggtgc ccatcctggc ctccattgtg ctcaacttca
     1861 tcctcttcat caatatcgtc cgggtgctcg ccaccaagct gcgggagacc aacgccggcc
     1921 ggtgtgacac acggcagcag taccggaagc tgctcaaatc cacgctggtg ctcatgcccc
     1981 tctttggcgt ccactacatt gtcttcatgg ccacaccata caccgaggtc tcagggacgc
     2041 tctggcaagt ccagatgcac tatgagatgc tcttcaactc cttccaggga ttttttgtcg
     2101 caatcatata ctgtttctgc aacggcgagg tacaagctga gatcaagaaa tcttggagcc
     2161 gctggacact ggcactggac ttcaagcgaa aggcacgcag cgggagcagc agctatagct
     2221 acggccccat ggtgtcccac acaagtgtga ccaatgtcgg cccccgtgtg ggactcggcc
     2281 tgcccctcag cccccgccta ctgcccactg ccaccaccaa cggccaccct cagctgcctg
     2341 gccatgccaa gccagggacc ccagccctgg agaccctcga gaccacacca cctgccatgg
     2401 ctgctcccaa ggacgatggg ttcctcaacg gctcctgctc aggcctggac gaggaggcct
     2461 ctgggcctga gcggccacct gccctgctac aggaagagtg ggagacagtc atgtgatcta
     2521 gaagttgtct cctcctgcac tgactgactg atacaatcga tttctggatc cgcaggcctc
     2581 tgctagcttg actgactgag atacagcgta ccttcagctc acagacatga taagatacat
     2641 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     2701 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
     2761 caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa
     2821 gtaaaacctc tacaaatgtg gtattggccc atctctatcg gtatcgtagc ataacccctt
     2881 ggggcctcta aacgggtctt gaggggtttt ttgtgcccct cgggccggat tgctatctac
     2941 cggcattggc gcagaaaaaa atgcctgatg cgacgctgcg cgtcttatac tcccacatat
     3001 gccagattca gcaacggata cggcttcccc aacttgccca cttccatacg tgtcctcctt
     3061 accagaaatt tatccttaag gtcgtcagct atcctgcagg cgatctctcg atttcgatca
     3121 agacattcct ttaatggtct tttctggaca ccactagggg tcagaagtag ttcatcaaac
     3181 tttcttccct ccctaatctc attggttacc ttgggctatc gaaacttaat taaccagtca
     3241 agtcagctac ttggcgagat cgacttgtct gggtttcgac tacgctcaga attgcgtcag
     3301 tcaagttcga tctggtcctt gctattgcac ccgttctccg attacgagtt tcatttaaat
     3361 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     3421 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     3481 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     3541 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     3601 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     3661 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     3721 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     3781 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     3841 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     3901 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     3961 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     4021 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     4081 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     4141 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     4201 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcattta aatttccgaa
     4261 ctctccaagg ccctcgtcgg aaaatcttca aacctttcgt ccgatccatc ttgcaggcta
     4321 cctctcgaac gaactatcgc aagtctcttg gccggccttg cgccttggct attgcttggc
     4381 agcgcctatc gccaggtatt actccaatcc cgaatatccg agatcgggat cacccgagag
     4441 aagttcaacc tacatcctca atcccgatct atccgagatc cgaggaatat cgaaatcggg
     4501 gcgcgcctgg cctccgcgcc gggttttggc gcctcccgcg ggcgcccccc tcgtcacggc
     4561 gagcgctgcc acgtcagacg aagggcgcag gagcgtcctg atccttccgc ccggacgctc
     4621 aggacagcgg cccgctgctc ataagactcg gccttagaac cccagtatca gcagaaggac
     4681 attttaggac gggacttggg tgactctagg gcactggttt tctttccaga gagcggaaca
     4741 ggcgaggaaa agtagtccct tctcggcgat tctgcggagg gatctccgtg gggcggtgaa
     4801 cgccgatgat tatataagga cgcgccgggt gtggcacagc tagttccgtc gcagccggga
     4861 tttgggtcgc ggttcttgtt tgtggatcgc tgtgatcgtc acttggtgag tagcgggctg
     4921 ctgggctggc cggggctttc gtggccgccg ggccgctcgg tgggacggaa gcgtgtggag
     4981 agaccgccaa gggctgtagt ctgggtccgc gagcaaggtt gccctgaact gggggttggg
     5041 gggagcgcag caaaatggcg gctgttcccg agtcttgaat ggaagacgct tgtgaggcgg
     5101 gctgtgaggt cgttgaaaca aggtgggggg catggtgggc ggcaagaacc caaggtcttg
     5161 agcccttcgc taatgcggga aagctcttat tcgggtgaga tgggctgggc accatctggg
     5221 gaccctgacg tgaagtttgt cactgactgg agaactcggt ttgtcgtctg ttgcgggggc
     5281 ggcagttatg gcggtgccgt tgggcagtgc acccgtacct ttgggagcgc gcgccctcgt
     5341 cgtgtcgtga cgtcacccgt tctgttggct tataatgcag ggtggggcca cctgccggta
     5401 ggtgtgcggt aggcttttct ccgtcgcagg acgcagggtt cgggcctagg gtaggctctc
     5461 ctgaatcgac aggcgccgga cctctggtga ggggagggat aagtgaggcg tcagtttctt
     5521 tggtcggttt tatgtaccta tcttcttaag tagctgaagc tccggttttg aactatgcgc
     5581 tcggggttgg cgagtgtgtt ttgtgaagtt ttttaggcac cttttgaaat gtaatcattt
     5641 gggtcaatat gtaattttca gtgttagact tgtaaattgt ccgctaaatt ctggccgttt
     5701 ttggcttttt tgttagacaa catgggtaaa aagcctgaac tcaccgcgac gtctgtcgag
     5761 aagtttctga tcgaaaagtt cgacagcgtc tccgacctga tgcagctctc ggagggcgaa
     5821 gaatctcgtg ctttcagctt cgatgtagga gggcgtggat atgtcctgcg ggtaaatagc
     5881 tgcgccgatg gtttctacaa agatcgttat gtttatcggc actttgcatc cgccgcgctc
     5941 ccgattccgg aagtgcttga cattggggag ttcagcgaga gcctgaccta ttgcatctcc
     6001 cgccgtgcac agggtgtcac gttgcaagac ctgcctgaaa ccgaactgcc cgctgttctg
     6061 cagccggtcg cggaggcaat ggatgccatc gctgccgccg atcttagcca gacgagcggg
     6121 ttcggcccat tcggaccgca aggaatcggt caatacacta catggcgtga tttcatatgc
     6181 gcgattgctg atccccatgt gtatcactgg caaactgtga tggacgacac cgtcagtgcg
     6241 tccgtcgcgc aggctctcga tgagctgatg ctttgggccg aggactgccc cgaagtccgg
     6301 cacctcgtgc acgcggattt cggctccaac aatgtcctga cggacaatgg ccgcataaca
     6361 gcggtcattg actggagcga ggcgatgttc ggggattccc aatacgaggt cgccaacatc
     6421 ttcttctgga ggccgtggtt ggcttgtatg gagcagcaga cgcgctactt cgagcggagg
     6481 catccggagc ttgcaggatc gccccggctc cgggcgtata tgctccgcat tggtcttgac
     6541 caactctatc agagcttggt tgacggcaat ttcgatgatg cagcttgggc gcagggtcga
     6601 tgcgacgcaa tcgtccgatc cggagccggg actgtcgggc gtacacaaat cgcccgcaga
     6661 agcgccgccg tctggaccga tggctgtgta gaagtactcg ccgatagtgg aaaccgacgc
     6721 cccagcactc gtccgagggc aaaggaataa gctagtatgt aagcctagtc ttagataata
     6781 aaatcgctat ccatcgaaga tggatgtgtg ttggtttttt gtgtgtgtaa cgctaggcgc
     6841 gcctggtgta ccgagaacga tcctctcagt gcgagtctcg acgatccata tcgttgcttg
     6901 gcagtcagcc agtcggaatc cagcttggga cccaggaagt ccaatcgtca gatattgtac
     6961 tcaagcctgg tcacggcagc gtaccgatct gtttaaacct agatattgat agtctgatcg
     7021 gtcaacgtat aatcgagtcc tagcttttgc aaacatctat caagagacag gatcagcagg
     7081 aggctttcgc atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     7141 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     7201 gttgggtgcg cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     7261 ttttcgcccc gaagaacgct ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     7321 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     7381 gaatgacttg gttgagtatt caccagtcac agaaaagcat cttacggatg gcatgacagt
     7441 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     7501 gacaacgatt ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     7561 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     7621 caccacgatg cctgtagcaa tggcaacaac cttgcgtaaa ctattaactg gcgaactact
     7681 tactctagct tcccggcaac agttgataga ctggatggag gcggataaag ttgcaggacc
     7741 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     7801 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     7861 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     7921 gataggtgcc tcactgatta agcattggta accgattcta ggtgcattgg cgcagaaaaa
     7981 aatgcctgat gcgacgctgc gcgtcttata ctcccacata tgccagattc agcaacggat
     8041 acggcttccc caacttgccc acttccatac gtgtcctcct taccagaaat ttatccttaa
     8101 gatcccgaat cgtttaaact cgactctggc tctatcgaat ctccgtcgtt tcgagcttac
     8161 gcgaacagcc gtggcgctca tttgctcgtc gggcatcgaa tctcgtcagc tatcgtcagc
     8221 ttaccttttt ggcagcgatc gcggctcccg acatcttgga ccattagctc cacaggtatc
     8281 ttcttccctc tagtggtcat aacagcagct tcagctacct ctcaattcaa aaaacccctc
     8341 aagacccgtt tagaggcccc aaggggttat gctatcaatc gttgcgttac acacacaaaa
     8401 aaccaacaca catccatctt cgatggatag cgattttatt atctaactgc tgatcgagtg
     8461 tagccagatc ta
//
