LOCUS       sequence-481980-        6948 bp    DNA     circular SYN 05-AUG-2025
DEFINITION  Express RPS6KA1-NanoLuc 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 6948)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6948)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 5, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: RPS6KA1-NanoLuc Fusion Vector
FEATURES             Location/Qualifiers
     source          1..6948
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     34..54
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          219..351
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     334..353
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     395..414
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        395..413
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2653..3165
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     primer_bind     complement(3253..3271)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3308..3429
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3319..3338)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3373..3392
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3674..4031
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3674..3865
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3697..3717)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3882..4017
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4046..4093
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4125..4919
                     /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(4179..4198)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4789..4808
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4983..5031
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(5270..5858)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5350..5369)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5974..6257
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
     enhancer        6448..6827
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(6828..6948,1..83)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
ORIGIN
        1 ttccaaaatg tcgtaataac cccgccccgt tgacgcaaat gggcggtagg cgtgtacggt
       61 gggaggtcta tataagcaga gctggtttag tgaaccgtca gatcactaga agctttattg
      121 cggtagttta tcacagttaa attgctaacg cagtcagtgc ttctgacaca acagtctcga
      181 acttaagctg cagaagttgg tcgtgaggca ctgggcaggt aagtatcaag gttacaagac
      241 aggtttaagg agaccaatag aaactgggct tgtcgagaca gagaagactc ttgcgtttct
      301 gataggcacc tattggtctt actgacatcc actttgcctt tctctccaca ggtgtccact
      361 cccagttcaa ttacagctct taaggctaga gtattaatac gactcactat agggctagcg
      421 atcgccatgc cgctcgccca gctcaaggag ccctggccgc tcatggagct agtgccgctg
      481 gacccggaga atggacagac ctcaggggaa gaagctggac ttcagccgtc caaggatgag
      541 ggcgtcctca aggagatctc catcacgcac cacgtcaagg ctggctctga gaaggctgat
      601 ccatcccatt tcgagctcct caaggttctg ggccagggat cctttggcaa agtcttcctg
      661 gtgcggaaag tcacccggcc tgacagtggg cacctgtatg ctatgaaggt gctgaagaag
      721 gcaacgctga aagtacgtga ccgcgtccgg accaagatgg agagagacat cctggctgat
      781 gtaaatcacc cattcgtggt gaagctgcac tatgccttcc agaccgaggg caagctctat
      841 ctcattctgg acttcctgcg tggtggggac ctcttcaccc ggctctcaaa agaggtgatg
      901 ttcacggagg aggatgtgaa gttttacctg gccgagctgg ctctgggcct ggatcacctg
      961 cacagcctgg gtatcattta cagagacctc aagcctgaga acatccttct ggatgaggag
     1021 ggccacatca aactcactga ctttggcctg agcaaagagg ccattgacca cgagaagaag
     1081 gcctattctt tctgcgggac agtggagtac atggcccctg aggtcgtcaa ccgccagggc
     1141 cactcccata gtgcggactg gtggtcctat ggggtgttga tgtttgagat gctgacgggc
     1201 tccctgccct tccaggggaa ggaccggaag gagaccatga cactgattct gaaggcgaag
     1261 ctaggcatgc cccagtttct gagcactgaa gcccagagcc tcttgcgggc cctgttcaag
     1321 cggaatcctg ccaaccggct cggctccggc cctgatgggg cagaggaaat caagcggcat
     1381 gtcttctact ccaccattga ctggaataag ctataccgtc gtgagatcaa gccacccttc
     1441 aagccagcag tggctcagcc tgatgacacc ttctactttg acaccgagtt cacgtcccgc
     1501 acacccaagg attccccagg catccccccc agcgctgggg cccatcagct gttccggggc
     1561 ttcagcttcg tggccaccgg cctgatggaa gacgacggca agcctcgtgc cccgcaggca
     1621 cccctgcact cggtggtaca gcaactccat gggaagaacc tggtttttag tgacggctac
     1681 gtggtaaagg agacaattgg tgtgggctcc tactctgagt gcaagcgctg tgtccacaag
     1741 gccaccaaca tggagtatgc tgtcaaggtc attgataaga gcaagcggga tccttcagaa
     1801 gagattgaga ttcttctgcg gtatggccag caccccaaca tcatcactct gaaagatgtg
     1861 tatgatgatg gcaaacacgt gtacctggtg acagagctga tgcggggtgg ggagctgctg
     1921 gacaagatcc tgcggcagaa gttcttctca gagcgggagg ccagctttgt cctgcacacc
     1981 attggcaaaa ctgtggagta tctgcactca cagggggttg tgcacaggga cctgaagccc
     2041 agcaacatcc tgtatgtgga cgagtccggg aatcccgagt gcctgcgcat ctgtgacttt
     2101 ggttttgcca aacagctgcg ggctgagaat gggctcctca tgacaccttg ctacacagcc
     2161 aactttgtgg cgcctgaggt gctgaagcgc cagggctacg atgaaggctg cgacatctgg
     2221 agcctgggca ttctgctgta caccatgctg gcaggatata ctccatttgc caacggtccc
     2281 agtgacacac cagaggaaat cctaacccgg atcggcagtg ggaagtttac cctcagtggg
     2341 ggaaattgga acacagtttc agagacagcc aaggacctgg tgtccaagat gctacacgtg
     2401 gatccccacc agcgcctcac agctaagcag gttctgcagc atccatgggt cacccagaaa
     2461 gacaagcttc cccaaagcca gctgtcccac caggacctac agcttgtgaa gggagccatg
     2521 gctgccacgt actccgcact caacagctcc aagcccaccc cccagctgaa gcccatcgag
     2581 tcatccatcc tggcccagcg gcgagtgagg aagttgccat ccaccaccct ggtttctctc
     2641 ggctcgagcg gcgtcttcac actcgaagat ttcgttgggg actggcgaca gacagccggc
     2701 tacaacctgg accaagtcct tgaacaggga ggtgtgtcca gtttgtttca gaatctcggg
     2761 gtgtccgtaa ctccgatcca aaggattgtc ctgagcggtg aaaatgggct gaagatcgac
     2821 atccatgtca tcatcccgta tgaaggtctg agcggcgacc aaatgggcca gatcgaaaaa
     2881 atttttaagg tggtgtaccc tgtggatgat catcacttta aggtgatcct gcactatggc
     2941 acactggtaa tcgacggggt tacgccgaac atgatcgact atttcggacg gccgtatgaa
     3001 ggcatcgccg tgttcgacgg caaaaagatc actgtaacag ggaccctgtg gaacggcaac
     3061 aaaattatcg acgagcgcct gatcaacccc gacggctccc tgctgttccg agtaaccatc
     3121 aacggagtga ccggctggcg gctgtgcgaa cgcattctgg cgtaaggccg cgactctaga
     3181 gtcgacctgc aggcatgcaa gctgatccgg ctgctaacaa agcccgaaag gaagctgagt
     3241 tggctgctgc caccgctgag caataactag cataacccct tggggcggcc gcttcgagca
     3301 gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa
     3361 tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat
     3421 aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggagatgtgg
     3481 gaggtttttt taagcaagta aaacctctac aaatgtggta aaatcgaatt ttaacaaaat
     3541 attaacgctt acaatttcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     3601 caccgcatac gcggatctgc gcagcaccat ggcctgaaat aacctctgaa agaggaactt
     3661 ggttaggtac cttctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg
     3721 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3781 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3841 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     3901 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     3961 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     4021 gcttttgcaa aaagcttaat taactgttga caattaatca tcggcatagt atatcggcat
     4081 agtataatac gacaaggtga ggaactaaac ccaggaggca gatcatgatt gaacaagatg
     4141 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     4201 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     4261 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
     4321 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     4381 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     4441 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     4501 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     4561 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     4621 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg
     4681 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     4741 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     4801 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     4861 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     4921 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgatggcc
     4981 gcaataaaat atctttattt tcattacatc tgtgtgttgg ttttttgtgt gaatcgatag
     5041 cgataaggat cctctttgcg cttgcgtttt cccttgtcca gatagcccag tagctgacat
     5101 tcatccgggg tcagcaccgt ttctgcggac tggctttcta cccggtatca gctcactcaa
     5161 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     5221 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     5281 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     5341 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     5401 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     5461 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     5521 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     5581 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     5641 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     5701 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     5761 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     5821 gcaagcagca gattacgcgc agaaaaaaag gatttcaaga agatcctttg atcttttcta
     5881 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     5941 caaaaaggat cttcacctag atccttttat agtccggaaa tacaggaacg cacgctggat
     6001 ggcccttcgc tgggatggtg aaaccatgaa aaatggcagc ttcagtggat taagtggggg
     6061 taatgtggcc tgtaccctct ggttgcatag gtattcatac ggttaaaatt tatcaggcgc
     6121 gattgcggca gtttttcggg tggtttgttg ccatttttac ctgtctgctg ccgtgatcgc
     6181 gctgaacgcg ttttagcggt gcgtacaatt aagggattat ggtaaatcca cttactgtct
     6241 gccctcgtag ccatcgagat aaaccgcagt actccggcca cgatgcgtcc ggcgtagagg
     6301 atcgagatct tcaatattgg ccattagcca tattattcat tggttatata gcataaatca
     6361 atattggcta ttggccattg catacgttgt atctatatca taatatgtac atttatattg
     6421 gctcatgtcc aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat
     6481 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
     6541 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
     6601 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
     6661 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg
     6721 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact
     6781 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
     6841 ggcagtacac caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
     6901 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggact
//
