LOCUS       sequence-478505-        7230 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Express NanoLuc-VDR Fusion Protein in Mammalian Cells under a CMV 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7230)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7230)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 7, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: NanoLuc-VDR Fusion Vector
FEATURES             Location/Qualifiers
     source          1..7230
                     /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(2993..3011)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3048..3169
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3059..3078)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3113..3132
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3414..3771
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3414..3605
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3437..3457)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3622..3757
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3802..4839
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             4818..4826
                     /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    4863..4911
                     /note="synthetic polyadenylation signal"
     promoter        5067..5171
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5172..5240,5241..6032)
                     /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(5390..5409)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(6190..6778)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6270..6289)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6894..7177
                     /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 cgatcgccat ggaggcaatg gcggccagca
     1621 cttccctgcc tgaccctgga gactttgacc ggaacgtgcc ccggatctgt ggggtgtgtg
     1681 gagaccgagc cactggcttt cacttcaatg ctatgacctg tgaaggctgc aaaggcttct
     1741 tcaggcgaag catgaagcgg aaggcactat tcacctgccc cttcaacggg gactgccgca
     1801 tcaccaagga caaccgacgc cactgccagg cctgccggct caaacgctgt gtggacatcg
     1861 gcatgatgaa ggagttcatt ctgacagatg aggaagtgca gaggaagcgg gagatgatcc
     1921 tgaagcggaa ggaggaggag gccttgaagg acagtctgcg gcccaagctg tctgaggagc
     1981 agcagcgcat cattgccata ctgctggacg cccaccataa gacctacgac cccacctact
     2041 ccgacttctg ccagttccgg cctccagttc gtgtgaatga tggtggaggg agccatcctt
     2101 ccaggcccaa ctccagacac actcccagct tctctgggga ctcctcctcc tcctgctcag
     2161 atcactgtat cacctcttca gacatgatgg actcgtccag cttctccaat ctggatctga
     2221 gtgaagaaga ttcagatgac ccttctgtga ccctagagct gtcccagctc tccatgctgc
     2281 cccacctggc tgacctggtc agttacagca tccaaaaggt cattggcttt gctaagatga
     2341 taccaggatt cagagacctc acctctgagg accagatcgt actgctgaag tcaagtgcca
     2401 ttgaggtcat catgttgcgc tccaatgagt ccttcaccat ggacgacatg tcctggacct
     2461 gtggcaacca agactacaag taccgcgtca gtgacgtgac caaagccgga cacagcctgg
     2521 agctgattga gcccctcatc aagttccagg tgggactgaa gaagctgaac ttgcatgagg
     2581 aggagcatgt cctgctcatg gccatctgca tcgtctcccc agatcgtcct ggggtgcagg
     2641 acgccgcgct gattgaggcc atccaggacc gcctgtccaa cacactgcag acgtacatcc
     2701 gctgccgcca cccgcccccg ggcagccacc tgctctatgc caagatgatc cagaagctag
     2761 ccgacctgcg cagcctcaat gaggagcact ccaagcagta ccgctgcctc tccttccagc
     2821 ctgagtgcag catgaagcta acgccccttg tgctcgaagt gtttggcaat gagatctccg
     2881 tttaaacgaa ttcgggctcg gtacccgggg atcctctaga gtcgacctgc aggcatgcaa
     2941 gctgatccgg ctgctaacaa agcccgaaag gaagctgagt tggctgctgc caccgctgag
     3001 caataactag cataacccct tggggcggcc gcttcgagca gacatgataa gatacattga
     3061 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     3121 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
     3181 ttgcattcat tttatgtttc aggttcaggg ggagatgtgg gaggtttttt taagcaagta
     3241 aaacctctac aaatgtggta aaatcgaatt ttaacaaaat attaacgctt acaatttcct
     3301 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac gcggatctgc
     3361 gcagcaccat ggcctgaaat aacctctgaa agaggaactt ggttaggtac cttctgaggc
     3421 ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     3481 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     3541 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     3601 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     3661 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     3721 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcgat
     3781 tcttctgaca ctagcgccac catgaagaag cccgaactca ccgctaccag cgttgaaaaa
     3841 tttctcatcg agaagttcga cagtgtgagc gacctgatgc agttgtcgga gggcgaagag
     3901 agccgagcct tcagcttcga tgtcggcgga cgcggctatg tactgcgggt gaatagctgc
     3961 gctgatggct tctacaaaga ccgctacgtg taccgccact tcgccagcgc tgcactaccc
     4021 atccccgaag tgttggacat cggcgagttc agcgagagcc tgacatactg catcagtaga
     4081 cgcgcccaag gcgttactct ccaagacctc cccgaaacag agctgcctgc tgtgttacag
     4141 cctgtcgccg aagctatgga tgctattgcc gccgccgacc tcagtcaaac cagcggcttc
     4201 ggcccattcg ggccccaagg catcggccag tacacaacct ggcgggattt catttgcgcc
     4261 attgctgatc cccatgtcta ccactggcag accgtgatgg acgacaccgt gtccgccagc
     4321 gtagctcaag ccctggacga actgatgctg tgggccgaag actgtcccga ggtgcgccac
     4381 ctcgtccatg ccgacttcgg cagcaacaac gtcctgaccg acaacggccg catcaccgcc
     4441 gtaatcgact ggtccgaagc tatgttcggg gacagtcagt acgaggtggc caacatcttc
     4501 ttctggcggc cctggctggc ttgcatggag cagcagactc gctacttcga gcgccggcat
     4561 cccgagctgg ccggcagccc tcgtctgcga gcctacatgc tgcgcatcgg cctggatcag
     4621 ctctaccaga gcctcgtgga cggcaacttc gacgatgctg cctgggctca aggccgctgc
     4681 gatgccatcg tccgcagcgg ggccggcacc gtcggtcgca cacaaatcgc tcgccggagc
     4741 gcagccgtat ggaccgacgg ctgcgtcgag gtgctggccg acagcggcaa ccgccggccc
     4801 agtacacgac cgcgcgctaa ggaggtaggt cgagtttaga ctctagccat cacgatggcc
     4861 gcaataaaat atctttattt tcattacatc tgtgtgttgg ttttttgtgt gaatcgatag
     4921 cgataaggat cctctttgcg cttgcgtttt cccttgtcca gatagcccag tagctgacat
     4981 tcatccgggg tcagcaccgt ttctgcggac tggctttcta cgtaatggtt tcttagacgt
     5041 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     5101 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     5161 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     5221 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     5281 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     5341 gttttcgccc cgaagaacgt tttccaatga tgagcacttt caaagttctg ctatgtggcg
     5401 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     5461 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     5521 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     5581 tgacaactat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     5641 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     5701 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     5761 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     5821 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     5881 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     5941 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     6001 agataggtgc ctcactgatt aagcattggt aattcgaaat gaccgaccaa gcgacgccca
     6061 accggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     6121 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     6181 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     6241 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     6301 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     6361 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     6421 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     6481 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     6541 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     6601 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     6661 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     6721 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatttcaaga
     6781 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     6841 gattttggtc atgagattat caaaaaggat cttcacctag atccttttat agtccggaaa
     6901 tacaggaacg cacgctggat ggcccttcgc tgggatggtg aaaccatgaa aaatggcagc
     6961 ttcagtggat taagtggggg taatgtggcc tgtaccctct ggttgcatag gtattcatac
     7021 ggttaaaatt tatcaggcgc gattgcggca gtttttcggg tggtttgttg ccatttttac
     7081 ctgtctgctg ccgtgatcgc gctgaacgcg ttttagcggt gcgtacaatt aagggattat
     7141 ggtaaatcca cttactgtct gccctcgtag ccatcgagat aaaccgcagt actccggcca
     7201 cgatgcgtcc ggcgtagagg atcgagatct
//
