LOCUS       sequence-473388-        7204 bp    DNA     circular SYN 14-JUL-2025
DEFINITION  ExpressVDR-NanoLuc(R) 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 7204)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 7204)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: VDR-NanoLuc Fusion Vector
FEATURES             Location/Qualifiers
     source          1..7204
                     /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             2367..2879
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     primer_bind     complement(2967..2985)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3022..3143
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3033..3052)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3087..3106
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3388..3745
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3388..3579
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3411..3431)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3596..3731
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3776..4813
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             4792..4800
                     /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    4837..4885
                     /note="synthetic polyadenylation signal"
     promoter        5041..5145
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5146..5214,5215..6006)
                     /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(5364..5383)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(6164..6752)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6244..6263)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6868..7151
                     /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 ggctagcgat cgccatggag gcaatggcgg
     1081 ccagcacttc cctgcctgac cctggagact ttgaccggaa cgtgccccgg atctgtgggg
     1141 tgtgtggaga ccgagccact ggctttcact tcaatgctat gacctgtgaa ggctgcaaag
     1201 gcttcttcag gcgaagcatg aagcggaagg cactattcac ctgccccttc aacggggact
     1261 gccgcatcac caaggacaac cgacgccact gccaggcctg ccggctcaaa cgctgtgtgg
     1321 acatcggcat gatgaaggag ttcattctga cagatgagga agtgcagagg aagcgggaga
     1381 tgatcctgaa gcggaaggag gaggaggcct tgaaggacag tctgcggccc aagctgtctg
     1441 aggagcagca gcgcatcatt gccatactgc tggacgccca ccataagacc tacgacccca
     1501 cctactccga cttctgccag ttccggcctc cagttcgtgt gaatgatggt ggagggagcc
     1561 atccttccag gcccaactcc agacacactc ccagcttctc tggggactcc tcctcctcct
     1621 gctcagatca ctgtatcacc tcttcagaca tgatggactc gtccagcttc tccaatctgg
     1681 atctgagtga agaagattca gatgaccctt ctgtgaccct agagctgtcc cagctctcca
     1741 tgctgcccca cctggctgac ctggtcagtt acagcatcca aaaggtcatt ggctttgcta
     1801 agatgatacc aggattcaga gacctcacct ctgaggacca gatcgtactg ctgaagtcaa
     1861 gtgccattga ggtcatcatg ttgcgctcca atgagtcctt caccatggac gacatgtcct
     1921 ggacctgtgg caaccaagac tacaagtacc gcgtcagtga cgtgaccaaa gccggacaca
     1981 gcctggagct gattgagccc ctcatcaagt tccaggtggg actgaagaag ctgaacttgc
     2041 atgaggagga gcatgtcctg ctcatggcca tctgcatcgt ctccccagat cgtcctgggg
     2101 tgcaggacgc cgcgctgatt gaggccatcc aggaccgcct gtccaacaca ctgcagacgt
     2161 acatccgctg ccgccacccg cccccgggca gccacctgct ctatgccaag atgatccaga
     2221 agctagccga cctgcgcagc ctcaatgagg agcactccaa gcagtaccgc tgcctctcct
     2281 tccagcctga gtgcagcatg aagctaacgc cccttgtgct cgaagtgttt ggcaatgaga
     2341 tctccgtttc tctcggctcg agcggcgtct tcacactcga agatttcgtt ggggactggc
     2401 gacagacagc cggctacaac ctggaccaag tccttgaaca gggaggtgtg tccagtttgt
     2461 ttcagaatct cggggtgtcc gtaactccga tccaaaggat tgtcctgagc ggtgaaaatg
     2521 ggctgaagat cgacatccat gtcatcatcc cgtatgaagg tctgagcggc gaccaaatgg
     2581 gccagatcga aaaaattttt aaggtggtgt accctgtgga tgatcatcac tttaaggtga
     2641 tcctgcacta tggcacactg gtaatcgacg gggttacgcc gaacatgatc gactatttcg
     2701 gacggccgta tgaaggcatc gccgtgttcg acggcaaaaa gatcactgta acagggaccc
     2761 tgtggaacgg caacaaaatt atcgacgagc gcctgatcaa ccccgacggc tccctgctgt
     2821 tccgagtaac catcaacgga gtgaccggct ggcggctgtg cgaacgcatt ctggcgtaag
     2881 gccgcgactc tagagtcgac ctgcaggcat gcaagctgat ccggctgcta acaaagcccg
     2941 aaaggaagct gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc
     3001 ggccgcttcg agcagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa
     3061 tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
     3121 ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc
     3181 agggggagat gtgggaggtt tttttaagca agtaaaacct ctacaaatgt ggtaaaatcg
     3241 aattttaaca aaatattaac gcttacaatt tcctgatgcg gtattttctc cttacgcatc
     3301 tgtgcggtat ttcacaccgc atacgcggat ctgcgcagca ccatggcctg aaataacctc
     3361 tgaaagagga acttggttag gtaccttctg aggcggaaag aaccagctgt ggaatgtgtg
     3421 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     3481 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     3541 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     3601 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     3661 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     3721 ttttggaggc ctaggctttt gcaaaaagct cgattcttct gacactagcg ccaccatgaa
     3781 gaagcccgaa ctcaccgcta ccagcgttga aaaatttctc atcgagaagt tcgacagtgt
     3841 gagcgacctg atgcagttgt cggagggcga agagagccga gccttcagct tcgatgtcgg
     3901 cggacgcggc tatgtactgc gggtgaatag ctgcgctgat ggcttctaca aagaccgcta
     3961 cgtgtaccgc cacttcgcca gcgctgcact acccatcccc gaagtgttgg acatcggcga
     4021 gttcagcgag agcctgacat actgcatcag tagacgcgcc caaggcgtta ctctccaaga
     4081 cctccccgaa acagagctgc ctgctgtgtt acagcctgtc gccgaagcta tggatgctat
     4141 tgccgccgcc gacctcagtc aaaccagcgg cttcggccca ttcgggcccc aaggcatcgg
     4201 ccagtacaca acctggcggg atttcatttg cgccattgct gatccccatg tctaccactg
     4261 gcagaccgtg atggacgaca ccgtgtccgc cagcgtagct caagccctgg acgaactgat
     4321 gctgtgggcc gaagactgtc ccgaggtgcg ccacctcgtc catgccgact tcggcagcaa
     4381 caacgtcctg accgacaacg gccgcatcac cgccgtaatc gactggtccg aagctatgtt
     4441 cggggacagt cagtacgagg tggccaacat cttcttctgg cggccctggc tggcttgcat
     4501 ggagcagcag actcgctact tcgagcgccg gcatcccgag ctggccggca gccctcgtct
     4561 gcgagcctac atgctgcgca tcggcctgga tcagctctac cagagcctcg tggacggcaa
     4621 cttcgacgat gctgcctggg ctcaaggccg ctgcgatgcc atcgtccgca gcggggccgg
     4681 caccgtcggt cgcacacaaa tcgctcgccg gagcgcagcc gtatggaccg acggctgcgt
     4741 cgaggtgctg gccgacagcg gcaaccgccg gcccagtaca cgaccgcgcg ctaaggaggt
     4801 aggtcgagtt tagactctag ccatcacgat ggccgcaata aaatatcttt attttcatta
     4861 catctgtgtg ttggtttttt gtgtgaatcg atagcgataa ggatcctctt tgcgcttgcg
     4921 ttttcccttg tccagatagc ccagtagctg acattcatcc ggggtcagca ccgtttctgc
     4981 ggactggctt tctacgtaat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
     5041 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     5101 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     5161 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     5221 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     5281 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     5341 atgatgagca ctttcaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     5401 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     5461 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     5521 accatgagtg ataacactgc ggccaactta cttctgacaa ctatcggagg accgaaggag
     5581 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     5641 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     5701 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     5761 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     5821 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     5881 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     5941 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     6001 tggtaattcg aaatgaccga ccaagcgacg cccaaccggt atcagctcac tcaaaggcgg
     6061 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     6121 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     6181 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     6241 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     6301 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     6361 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     6421 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     6481 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     6541 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     6601 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     6661 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     6721 agcagattac gcgcagaaaa aaaggatttc aagaagatcc tttgatcttt tctacggggt
     6781 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     6841 ggatcttcac ctagatcctt ttatagtccg gaaatacagg aacgcacgct ggatggccct
     6901 tcgctgggat ggtgaaacca tgaaaaatgg cagcttcagt ggattaagtg ggggtaatgt
     6961 ggcctgtacc ctctggttgc ataggtattc atacggttaa aatttatcag gcgcgattgc
     7021 ggcagttttt cgggtggttt gttgccattt ttacctgtct gctgccgtga tcgcgctgaa
     7081 cgcgttttag cggtgcgtac aattaaggga ttatggtaaa tccacttact gtctgccctc
     7141 gtagccatcg agataaaccg cagtactccg gccacgatgc gtccggcgta gaggatcgag
     7201 atct
//
