LOCUS       sequence-471874-        6941 bp    DNA     circular SYN 14-JUL-2025
DEFINITION  Express NanoLuc(R)-HSP90AB1 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 6941)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 6941)
  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: NanoLuc-HSP90AB1 Fusion Vector
FEATURES             Location/Qualifiers
     source          1..6941
                     /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(3884..3902)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3939..4060
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3950..3969)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4004..4023
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4305..4662
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4305..4496
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4328..4348)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4513..4648
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4677..4724
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4756..5550
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4810..4829)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5420..5439
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5614..5662
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(5901..6489)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5981..6000)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6605..6888
                     /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 gcctgaggaa gtgcaccatg
     1621 gagaggagga ggtggagact tttgcctttc aggcagaaat tgcccaactc atgtccctca
     1681 tcatcaatac cttctattcc aacaaggaga ttttccttcg ggagttgatc tctaatgctt
     1741 ctgatgcctt ggacaagatt cgctatgaga gcctgacaga cccttcgaag ttggacagtg
     1801 gtaaagagct gaaaattgac atcatcccca accctcagga acgtaccctg actttggtag
     1861 acacaggcat tggcatgacc aaagctgatc tcataaataa tttgggaacc attgccaagt
     1921 ctggtactaa agcattcatg gaggctcttc aggctggtgc agacatctcc atgattgggc
     1981 agtttggtgt tggcttttat tctgcctact tggtggcaga gaaagtggtt gtgatcacaa
     2041 agcacaacga tgatgaacag tatgcttggg agtcttctgc tggaggttcc ttcactgtgc
     2101 gtgctgacca tggtgagccc attggcaggg gtaccaaagt gatcctccat cttaaagaag
     2161 atcagacaga gtacctagaa gagaggcggg tcaaagaagt agtgaagaag cattctcagt
     2221 tcataggcta tcccatcacc ctttatttgg agaaggaacg agagaaggaa attagtgatg
     2281 atgaggcaga ggaagagaaa ggtgagaaag aagaggaaga taaagatgat gaagaaaaac
     2341 ccaagatcga agatgtgggt tcagatgagg aggatgacag cggtaaggat aagaagaaga
     2401 aaactaagaa gatcaaagag aaatacattg atcaggaaga actaaacaag accaagccta
     2461 tttggaccag aaaccctgat gacatcaccc aagaggagta tggagaattc tacaagagcc
     2521 tcactaatga ctgggaagac cacttggcag tcaagcactt ttctgtagaa ggtcagttgg
     2581 aattcagggc attgctattt attcctcgtc gggctccctt tgaccttttt gagaacaaga
     2641 agaaaaagaa caacatcaaa ctctatgtcc gccgtgtgtt catcatggac agctgtgatg
     2701 agttgatacc agagtatctc aattttatcc gtggtgtggt tgactctgag gatctgcccc
     2761 tgaacatctc ccgagaaatg ctccagcaga gcaaaatctt gaaagtcatt cgcaaaaaca
     2821 ttgttaagaa gtgccttgag ctcttctctg agctggcaga agacaaggag aattacaaga
     2881 aattctatga ggcattctct aaaaatctca agcttggaat ccacgaagac tccactaacc
     2941 gccgccgcct gtctgagctg ctgcgctatc atacctccca gtctggagat gagatgacat
     3001 ctctgtcaga gtatgtttct cgcatgaagg agacacagaa gtccatctat tacatcactg
     3061 gtgagagcaa agagcaggtg gccaactcag cttttgtgga gcgagtgcgg aaacggggct
     3121 tcgaggtggt atatatgacc gagcccattg acgagtactg tgtgcagcag ctcaaggaat
     3181 ttgatgggaa gagcctggtc tcagttacca aggagggtct ggagctgcct gaggatgagg
     3241 aggagaagaa gaagatggaa gagagcaagg caaagtttga gaacctctgc aagctcatga
     3301 aagaaatctt agataagaag gttgagaagg tgacaatctc caatagactt gtgtcttcac
     3361 cttgctgcat tgtgaccagc acctacggct ggacagccaa tatggagcgg atcatgaaag
     3421 cccaggcact tcgggacaac tccaccatgg gctatatgat ggccaaaaag cacctggaga
     3481 tcaaccctga ccaccccatt gtggagacgc tgcggcagaa ggctgaggcc gacaagaatg
     3541 ataaggcagt taaggacctg gtggtgctgc tgtttgaaac cgccctgcta tcttctggct
     3601 tttcccttga ggatccccag acccactcca accgcatcta tcgcatgatc aagctaggtc
     3661 taggtattga tgaagatgaa gtggcagcag aggaacccaa tgctgcagtt cctgatgaga
     3721 tcccccctct cgagggcgat gaggatgcgt ctcgcatgga agaagtcgat gtttaaacga
     3781 attcgggctc ggtacccggg gatcctctag agtcgacctg caggcatgca agctgatccg
     3841 gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga gcaataacta
     3901 gcataacccc ttggggcggc cgcttcgagc agacatgata agatacattg atgagtttgg
     3961 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     4021 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     4081 ttttatgttt caggttcagg gggagatgtg ggaggttttt ttaagcaagt aaaacctcta
     4141 caaatgtggt aaaatcgaat tttaacaaaa tattaacgct tacaatttcc tgatgcggta
     4201 ttttctcctt acgcatctgt gcggtatttc acaccgcata cgcggatctg cgcagcacca
     4261 tggcctgaaa taacctctga aagaggaact tggttaggta ccttctgagg cggaaagaac
     4321 cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     4381 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     4441 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     4501 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     4561 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     4621 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagcttaa ttaactgttg
     4681 acaattaatc atcggcatag tatatcggca tagtataata cgacaaggtg aggaactaaa
     4741 cccaggaggc agatcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     4801 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     4861 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     4921 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     4981 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     5041 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     5101 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     5161 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     5221 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     5281 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     5341 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     5401 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     5461 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     5521 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     5581 aagcgacgcc caacctgcca tcacgatggc cgcaataaaa tatctttatt ttcattacat
     5641 ctgtgtgttg gttttttgtg tgaatcgata gcgataagga tcctctttgc gcttgcgttt
     5701 tcccttgtcc agatagccca gtagctgaca ttcatccggg gtcagcaccg tttctgcgga
     5761 ctggctttct acccggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     5821 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     5881 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     5941 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     6001 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     6061 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     6121 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     6181 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     6241 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     6301 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
     6361 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     6421 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     6481 ggatttcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     6541 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     6601 tagtccggaa atacaggaac gcacgctgga tggcccttcg ctgggatggt gaaaccatga
     6661 aaaatggcag cttcagtgga ttaagtgggg gtaatgtggc ctgtaccctc tggttgcata
     6721 ggtattcata cggttaaaat ttatcaggcg cgattgcggc agtttttcgg gtggtttgtt
     6781 gccattttta cctgtctgct gccgtgatcg cgctgaacgc gttttagcgg tgcgtacaat
     6841 taagggatta tggtaaatcc acttactgtc tgccctcgta gccatcgaga taaaccgcag
     6901 tactccggcc acgatgcgtc cggcgtagag gatcgagatc t
//
