LOCUS       sequence-479787-        6086 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Express NanoLuc-cMyc 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 6086)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6086)
  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            "Unpublished"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-cMyc Vector
FEATURES             Location/Qualifiers
     source          1..6086
                     /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"
     CDS             1599..2921
                     /codon_start=1
                     /product="human c-Myc proto-oncogene"
                     /label=c-Myc
                     /translation="MPLNVSFTNRNYDLDYDSVQPYFYCDEEENFYQQQQQSELQPPAP
                     SEDIWKKFELLPTPPLSPSRRSGLCSPSYVAVTPFSLRGDNDGGGGSFSTADQLEMVTE
                     LLGGDMVNQSFICDPDDETFIKNIIIQDCMWSGFSAAAKLVSEKLASYQAARKDSGSPN
                     PARGHSVCSTSSLYLQDLSAAASECIDPSVVFPYPLNDSSSPKSCASQDSSAFSPSSDS
                     LLSSTESSPQGSPEPLVLHEETPPTTSSDSEEEQEDEEEIDVVSVEKRQAPGKRSESGS
                     PSAGGHSKPPHSPLVLKRCHVSTHQHNYAAPPSTRKDYPAAKRVKLDSVRVLRQISNNR
                     KCTSPRSSDTEENVKRRTHNVLERQRRNELKRSFFALRDQIPELENNEKAPKVVILKKA
                     TAYILSVQAEEQKLISEEDLLRKRREQLKHKLEQLRNSCAV"
     primer_bind     complement(3029..3047)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3084..3205
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3095..3114)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3149..3168
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3450..3807
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3450..3641
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3473..3493)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3658..3793
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3822..3869
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3901..4695
                     /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(3955..3974)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4565..4584
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4759..4807
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(5046..5634)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5126..5145)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5750..6033
                     /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 gcccctcaac gttagcttca
     1621 ccaacaggaa ctatgacctc gactacgact cggtgcagcc gtatttctac tgcgacgagg
     1681 aggagaactt ctaccagcag cagcagcaga gcgagctgca gcccccggcg cccagcgagg
     1741 atatctggaa gaaattcgag ctgctgccca ccccgcccct gtcccctagc cgccgctccg
     1801 ggctctgctc gccctcctac gttgcggtca cacccttctc ccttcgggga gacaacgacg
     1861 gcggtggcgg gagcttctcc acggccgacc agctggagat ggtgaccgag ctgctgggag
     1921 gagacatggt gaaccagagt ttcatctgcg acccggacga cgagaccttc atcaaaaaca
     1981 tcatcatcca ggactgtatg tggagcggct tctcggccgc cgccaagctc gtctcagaga
     2041 agctggcctc ctaccaggct gcgcgcaaag acagcggcag cccgaacccc gcccgcggcc
     2101 acagcgtctg ctccacctcc agcttgtacc tgcaggatct gagcgccgcc gcctcagagt
     2161 gcatcgaccc ctcggtggtc ttcccctacc ctctcaacga cagcagctcg cccaagtcct
     2221 gcgcctcgca agactccagc gccttctctc cgtcctcgga ttctctgctc tcctcgacgg
     2281 agtcctcccc gcagggcagc cccgagcccc tggtgctcca tgaggagaca ccgcccacca
     2341 ccagcagcga ctctgaggag gaacaagaag atgaggaaga aatcgatgtt gtttctgtgg
     2401 aaaagaggca ggctcctggc aaaaggtcag agtctggatc accttctgct ggaggccaca
     2461 gcaaacctcc tcacagccca ctggtcctca agaggtgcca cgtctccaca catcagcaca
     2521 actacgcagc gcctccctcc actcggaagg actatcctgc tgccaagagg gtcaagttgg
     2581 acagtgtcag agtcctgaga cagatcagca acaaccgaaa atgcaccagc cccaggtcct
     2641 cggacaccga ggagaatgtc aagaggcgaa cacacaacgt cttggagcgc cagaggagga
     2701 acgagctaaa acggagcttt tttgccctgc gtgaccagat cccggagttg gaaaacaatg
     2761 aaaaggcccc caaggtagtt atccttaaaa aagccacagc atacatcctg tccgtccaag
     2821 cagaggagca aaagctcatt tctgaagagg acttgttgcg gaaacgacga gaacagttga
     2881 aacacaaact tgaacagcta cggaactctt gtgcggttta aacgaattcg ggctcggtac
     2941 ccggggatcc tctagagtcg acctgcaggc atgcaagctg atccggctgc taacaaagcc
     3001 cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg
     3061 gcggccgctt cgagcagaca tgataagata cattgatgag tttggacaaa ccacaactag
     3121 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     3181 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     3241 tcagggggag atgtgggagg tttttttaag caagtaaaac ctctacaaat gtggtaaaat
     3301 cgaattttaa caaaatatta acgcttacaa tttcctgatg cggtattttc tccttacgca
     3361 tctgtgcggt atttcacacc gcatacgcgg atctgcgcag caccatggcc tgaaataacc
     3421 tctgaaagag gaacttggtt aggtaccttc tgaggcggaa agaaccagct gtggaatgtg
     3481 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3541 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     3601 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     3661 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     3721 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     3781 ttttttggag gcctaggctt ttgcaaaaag cttaattaac tgttgacaat taatcatcgg
     3841 catagtatat cggcatagta taatacgaca aggtgaggaa ctaaacccag gaggcagatc
     3901 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     3961 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     4021 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     4081 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     4141 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     4201 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     4261 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     4321 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     4381 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     4441 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     4501 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     4561 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     4621 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     4681 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     4741 tgccatcacg atggccgcaa taaaatatct ttattttcat tacatctgtg tgttggtttt
     4801 ttgtgtgaat cgatagcgat aaggatcctc tttgcgcttg cgttttccct tgtccagata
     4861 gcccagtagc tgacattcat ccggggtcag caccgtttct gcggactggc tttctacccg
     4921 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     4981 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     5041 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     5101 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     5161 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     5221 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     5281 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     5341 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     5401 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     5461 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
     5521 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     5581 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatt tcaagaagat
     5641 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     5701 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttatagtc cggaaataca
     5761 ggaacgcacg ctggatggcc cttcgctggg atggtgaaac catgaaaaat ggcagcttca
     5821 gtggattaag tgggggtaat gtggcctgta ccctctggtt gcataggtat tcatacggtt
     5881 aaaatttatc aggcgcgatt gcggcagttt ttcgggtggt ttgttgccat ttttacctgt
     5941 ctgctgccgt gatcgcgctg aacgcgtttt agcggtgcgt acaattaagg gattatggta
     6001 aatccactta ctgtctgccc tcgtagccat cgagataaac cgcagtactc cggccacgat
     6061 gcgtccggcg tagaggatcg agatct
//
