LOCUS       sequence-481959-        7461 bp    DNA     circular SYN 04-AUG-2025
DEFINITION  Express NanoLuc-RAD21 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 7461)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7461)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 4, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: NanoLuc-RAD21 Vector
FEATURES             Location/Qualifiers
     source          1..7461
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    17..98
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     42..61
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(2215..2727)
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     promoter        complement(2766..2784)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2767..2784)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     enhancer        2904..3207
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     3362..3382
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        3415..3462
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3494..4153
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3560..3579)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      complement(4325..4913)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4405..4424)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5039..5087
                     /note="synthetic polyadenylation signal"
     CDS             complement(5111..5905)
                     /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(5222..5241)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5832..5851
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(6283..7461)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          6292..7230
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     complement(6315..6335)
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
ORIGIN
        1 aagacgcgtg acgtcggcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
       61 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
      121 ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca
      181 aatgtggtat ggctgattat gatccgcggc cgcaatacgt cgacgttatc agctgacttc
      241 gtacgagagc ctaggtgttt ctagaggatc cccgggtacc gagcccgaat tcgtttaaac
      301 tataatatgg aaccttggtc caggtgttgc gatgatgtca ctgtacggtt cttcctgtgt
      361 cagctcaata gcttgctgct ttttaagaac caagaagctg tagaactttg cggcagcttg
      421 ttttctgttc gtatttcgac ataactcaag caaactgata gattcagctc cagttttagc
      481 aagagcacgc tgaagaccat gaagcatctg ctgagtcctt ttgttccatc ttctttcttc
      541 ctgatcttga tcgccccctg atgcatcttc atcctcttcc tcttcatcat cttctttttc
      601 cttctctttc tccttctctt tttctggcag aagttctaac tctggtatta gctgacagat
      661 atttggaggt tcttctgggg gaagctctac aggtggtatt tccatctgct ctacctgctg
      721 aggaggcatc acaggctctg ggtcaatttg tccagctttt cgcttaactc cctgaggtgg
      781 tggtggaggc atagctgact catctatgtt tgttctgctg gcctccatca ctgactcctg
      841 gaggcggctt ggctcttcaa taatgggctc atcgataaca tcacgctgct gatgctgctg
      901 ttgctggtcc tctctaggaa cctctggatt ttcaaattct ttgaggaatt catccaaatt
      961 atctgcctct cctcctttcc tcctttttct aaggtcttct ggtacaagcg gtgtaagaca
     1021 gcgtgtaaag agcttcagta gtctgttatt ccacaaaggc tgagcaggta aagaaaacag
     1081 tttttctact cctcctgtct ctttccacat catcaatttc ttggtgggcg gtgccagatc
     1141 caaagtagta acaatatctg aataatcact aagttgggct ctaattgtct tgctatccaa
     1201 ctctttgaca ctgtcaacaa ttagcttcct cttcctcttg gcttttgttt ctttaacagt
     1261 tatatcaata ggctccaatg caaatgcttc ttcctcattt ggaacaagtg ttgtttgatc
     1321 agtcatggtt ggcattggtt caacgggatc cactgaatca ggactatcag gcccacccat
     1381 tgatacatta tcatcctcat ccatatcgtc atgtgcaggc tgctctggca acatcacccc
     1441 tgcctcagag agggcagggg gatcatcaaa gataccgcca tcattattac taataagttt
     1501 gtcatctaat attccaccat catttccttc tccaaaatta tcatccttat attgatcttc
     1561 atattctaaa tggttaattt tctcattcag attgctggtg ctctgttcag actctaatag
     1621 gaggttagaa gtagtagtgc ttactaacat gtcgtcatcc tcaaaagcac tgccttctct
     1681 cattatctca cgatcatcca ttccaaaatc accaaaatca ttttcttgta aaatactgat
     1741 gttcccaact tcttctctca tggttatctc ttccactcta ctctgattca agctgaactg
     1801 ctgggccaca tcgatgtcat ctaagtcagg cagtggctga tcaaagtcat gaaattcttc
     1861 aggtaaagta atggcattat aagctgcttc ccgattttcc tcaggcaggt caaccacacc
     1921 tggccgaaaa gccatcttta tcttaatgaa tgcttcatta cagtctgcaa gaaggtattt
     1981 ggctttcctg tgatagattc gaactactcc cagtaagaga tgtcctgatg tccgtaatgc
     2041 cattttcacc tttggtgaga tgatactctc cacgctgctc tctaaattac actcgaacac
     2101 atgggctttg gttagcttct tatcccaatg ggccgctagc caaattttgg ccagaggccc
     2161 tcttttactg agaacaaaat gtgcgtagaa catggcgatc gcgccgctcg agcccgccag
     2221 aatgcgttcg cacagccgcc agccggtcac tccgttgatg gttactcgga acagcaggga
     2281 gccgtcgggg ttgatcaggc gctcgtcgat aattttgttg ccgttccaca gggtccctgt
     2341 tacagtgatc tttttgccgt cgaacacggc gatgccttca tacggccgtc cgaaatagtc
     2401 gatcatgttc ggcgtaaccc cgtcgattac cagtgtgcca tagtgcagga tcaccttaaa
     2461 gtgatgatca tccacagggt acaccacctt aaaaattttt tcgatctggc ccatttggtc
     2521 gccgctcaga ccttcatacg ggatgatgac atggatgtcg atcttcagcc cattttcacc
     2581 gctcaggaca atcctttgga tcggagttac ggacaccccg agattctgaa acaaactgga
     2641 cacacctccc tgttcaagga cttggtccag gttgtagccg gctgtctgtc gccagtcccc
     2701 aacgaaatct tcgagtgtga agaccatggt gagcgctagc tgcaaaaaga acaagtagct
     2761 tgtattctat agtgtcacct aaatgggcga tctgacggtt cactaaacca gctctgctta
     2821 tataggcctc cctggcgcgc cactagttat taatagtaat caattacggg gtcattagtt
     2881 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
     2941 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
     3001 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
     3061 gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga cggtaaatgg
     3121 cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg gcagtacatc
     3181 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac caatgggcgt
     3241 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
     3301 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc cgccccgttg
     3361 acgcaaatgg gcggtaggcg tgtacggtgg gaggtcaaat cgaattctaa tactgttgac
     3421 aattaatcat cggcatagta tatcggcata gtataatacg acaaggtgag gaactaaacc
     3481 caggaggcag atcatggaga aaaaaatcac tggatatacc accgttgata tatcccaatg
     3541 gcatcgtaaa gaacattttg aggcatttca gtcagttgct caatgtacct ataaccagac
     3601 cgttcagctg gatattacgg cctttttaaa gaccgtaaag aaaaataagc acaagtttta
     3661 tccggccttt attcacattc ttgcccgcct gatgaatgct catccggaat tccgtatggc
     3721 aatgaaagac ggtgagctgg tgatatggga tagtgttcac ccttgttaca ccgttttcca
     3781 tgagcaaact gaaacgtttt catcgctctg gagtgaatac cacgacgatt tccggcagtt
     3841 tctacacata tattcgcaag atgtggcgtg ttacggtgaa aacctggcct atttccctaa
     3901 agggtttatt gagaatatgt ttttcgtctc agccaatccc tgggtgagtt tcaccagttt
     3961 tgatttaaac gtggccaata tggacaactt cttcgccccc gttttcacta tgggcaaata
     4021 ttatacgcaa ggcgacaagg tgctgatgcc gctggcgatt caggttcatc atgccgtctg
     4081 tgatggcttc catgtcggca gaatgcttaa tgaattacaa cagtactgcg atgagtggca
     4141 gggcggggcg taagcgggac tctggggttc gaaatgaccg accaagcgac gcccaaccgg
     4201 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     4261 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     4321 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     4381 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     4441 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     4501 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     4561 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     4621 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     4681 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     4741 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     4801 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     4861 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggattt caagaagatc
     4921 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     4981 tggtcatgag attatcaaaa aggatcttca cctagatcct tttatagtcc ggaatcgtca
     5041 cacaaaaaac caacacacag atgtaatgaa aataaagata ttttattgcg gccatcgtga
     5101 tggctagagt tcagaagaac tcgtcaagaa ggcgatagaa ggcgatgcgc tgcgaatcgg
     5161 gagcggcgat accgtaaagc acgaggaagc ggtcagccca ttcgccgcca agctcttcag
     5221 caatatcacg ggtagccaac gctatgtcct gatagcggtc cgccacaccc agccggccac
     5281 agtcgatgaa tccagaaaag cggccatttt ccaccatgat attcggcaag caggcatcgc
     5341 cgtgggtcac gacgagatcc tcgccgtcgg gcatgcgcgc cttgagcctg gcgaacagtt
     5401 cggctggcgc gagcccctga tgctcttcgt ccagatcatc ctgatcgaca agaccggctt
     5461 ccatccgagt acgtgctcgc tcgatgcgat gtttcgcttg gtggtcgaat gggcaggtag
     5521 ccggatcaag cgtatgcagc cgccgcattg catcagccat gatggatact ttctcggcag
     5581 gagcaaggtg agatgacagg agatcctgcc ccggcacttc gcccaatagc agccagtccc
     5641 ttcccgcttc agtgacaacg tcgagcacag ctgcgcaagg aacgcccgtc gtggccagcc
     5701 acgatagccg cgctgcctcg tcctgcagtt cattcagggc accggacagg tcggtcttga
     5761 caaaaagaac cgggcgcccc tgcgctgaca gccggaacac ggcggcatca gagcagccga
     5821 ttgtctgttg tgcccagtca tagccgaata gcctctccac ccaagcggcc ggagaacctg
     5881 cgtgcaatcc atcttgttca atcatggtgg catcgattcc cggtcaacat ccagtacacc
     5941 tccggcacac cggcttcccc aagggcgcca cgctctccca ccacaacatc ctcaacaacg
     6001 gctactgggt gggccgcacg gtcggctaca cggagcagga ccgggtgtgt ctgccggtgc
     6061 ccttctacca ctgcttcggc atggtcatgg gcaacttggg cgccacctcc cacgccgcct
     6121 gcatcgtcat cccggccccg tccttcgagc cggcggcgac gctggaggcg gtgcagcggg
     6181 agaggtgcac gtccctgtac ggcgtcccca cgaagccgaa ttccaccaca ctggactaga
     6241 tgcatgctcg acagtgtgat ggatatctgc agaattcggc tttcacgaca cctgaaatgg
     6301 aagaaaaaaa ctttgaacca ctgtctgagg cttgagaatg aaccaagatc caaactcaaa
     6361 aagggcaaat tccaaggaga attacatcaa gtgccaagct ggcctaactt cagtctccac
     6421 ccactcagtg tggggaaact ccatcgcata aaacccctcc ccccaaccta aagacgacgt
     6481 actccaaaag ctcgagaact aatcgaggtg cctggacggc gcccggtact ccgtggagtc
     6541 acatgaagcg acggctgagg acggaaaggc ccttttcctt tgtgtgggtg actcacccgc
     6601 ccgctctccc gagcgccgcg tcctccattt tgagctgcct gcagcagggc cgggaagcgg
     6661 ccatctttcc gctcacgcaa ctggtgccga ccgggccagc cttgccgccc agggcggggc
     6721 gatacacggc ggcgcgaggc caggcaccag agcaggccgg ccagcttgag actacccccg
     6781 tccgattctc ggtggccgcg ctcgcaggcc ccgcctcgcc gaacatgtgc gctgggacgc
     6841 acgggccccg tcgccgcccg cggccccaaa aaccgaaata ccagtgtgca gatcttggcc
     6901 cgcatttaca agactatctt gccagaaaaa aagcgtcgca gcaggtcatc aaaaatttta
     6961 aatggctaga gacttatcga aagcagcgag acaggcgcga aggtgccacc agattcgcac
     7021 gcggcggccc cagcgcccag gccaggcctc aactcaagca cgaggcgaag gggctcctta
     7081 agcgcaaggc ctcgaactct cccacccact tccaacccga agctcgggat caagaatcac
     7141 gtactgcagc caggtggaag taattcaagg cacgcaaggg ccataacccg taaagaggcc
     7201 aggcccgcgg gaaccacaca cggcacttac ctgtgttctg gcggcaaacc cgttgcgaaa
     7261 aagaacgttc acggcgacta ctgcacttat atacggttct cccccaccct cgggaaaaag
     7321 gcggagccag tacacgacat cactttccca gtttaccccg cgccaccttc tctaggcacc
     7381 ggttcaattg ccgacccctc cccccaactt ctcggggact gtgggcgatg tgcgctctgc
     7441 ccactgacgg gcaccggagc c
//
