LOCUS       sequence-480848-        7435 bp    DNA     circular SYN 28-JUL-2025
DEFINITION  Express RAD21-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 7435)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7435)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 28, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: RAD21-NanoLuc Vector
FEATURES             Location/Qualifiers
     source          1..7435
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     307..327
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        360..407
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             439..1098
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(505..524)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      complement(1270..1858)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1350..1369)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    1984..2032
                     /note="synthetic polyadenylation signal"
     CDS             complement(2056..2850)
                     /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(2167..2186)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2777..2796
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(3228..4406)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          3237..4175
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     complement(3260..3280)
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     polyA_signal    4423..4504
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     4448..4467
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(4681..5193)
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     promoter        complement(7146..7164)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(7147..7164)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     enhancer        join(7284..7435,1..152)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tggcagtaca tcaagtgtat catatgccaa gtccgccccc tattgacgtc aatgacggta
       61 aatggcccgc ctggcattat gcccagtaca tgaccttacg ggactttcct acttggcagt
      121 acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag tacaccaatg
      181 ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg
      241 ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaat aaccccgccc
      301 cgttgacgca aatgggcggt aggcgtgtac ggtgggaggt caaatcgaat tctaatactg
      361 ttgacaatta atcatcggca tagtatatcg gcatagtata atacgacaag gtgaggaact
      421 aaacccagga ggcagatcat ggagaaaaaa atcactggat ataccaccgt tgatatatcc
      481 caatggcatc gtaaagaaca ttttgaggca tttcagtcag ttgctcaatg tacctataac
      541 cagaccgttc agctggatat tacggccttt ttaaagaccg taaagaaaaa taagcacaag
      601 ttttatccgg cctttattca cattcttgcc cgcctgatga atgctcatcc ggaattccgt
      661 atggcaatga aagacggtga gctggtgata tgggatagtg ttcacccttg ttacaccgtt
      721 ttccatgagc aaactgaaac gttttcatcg ctctggagtg aataccacga cgatttccgg
      781 cagtttctac acatatattc gcaagatgtg gcgtgttacg gtgaaaacct ggcctatttc
      841 cctaaagggt ttattgagaa tatgtttttc gtctcagcca atccctgggt gagtttcacc
      901 agttttgatt taaacgtggc caatatggac aacttcttcg cccccgtttt cactatgggc
      961 aaatattata cgcaaggcga caaggtgctg atgccgctgg cgattcaggt tcatcatgcc
     1021 gtctgtgatg gcttccatgt cggcagaatg cttaatgaat tacaacagta ctgcgatgag
     1081 tggcagggcg gggcgtaagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     1141 accggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     1201 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     1261 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     1321 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     1381 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     1441 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     1501 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     1561 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     1621 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     1681 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     1741 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     1801 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatttcaaga
     1861 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     1921 gattttggtc atgagattat caaaaaggat cttcacctag atccttttat agtccggaat
     1981 cgtcacacaa aaaaccaaca cacagatgta atgaaaataa agatatttta ttgcggccat
     2041 cgtgatggct agagttcaga agaactcgtc aagaaggcga tagaaggcga tgcgctgcga
     2101 atcgggagcg gcgataccgt aaagcacgag gaagcggtca gcccattcgc cgccaagctc
     2161 ttcagcaata tcacgggtag ccaacgctat gtcctgatag cggtccgcca cacccagccg
     2221 gccacagtcg atgaatccag aaaagcggcc attttccacc atgatattcg gcaagcaggc
     2281 atcgccgtgg gtcacgacga gatcctcgcc gtcgggcatg cgcgccttga gcctggcgaa
     2341 cagttcggct ggcgcgagcc cctgatgctc ttcgtccaga tcatcctgat cgacaagacc
     2401 ggcttccatc cgagtacgtg ctcgctcgat gcgatgtttc gcttggtggt cgaatgggca
     2461 ggtagccgga tcaagcgtat gcagccgccg cattgcatca gccatgatgg atactttctc
     2521 ggcaggagca aggtgagatg acaggagatc ctgccccggc acttcgccca atagcagcca
     2581 gtcccttccc gcttcagtga caacgtcgag cacagctgcg caaggaacgc ccgtcgtggc
     2641 cagccacgat agccgcgctg cctcgtcctg cagttcattc agggcaccgg acaggtcggt
     2701 cttgacaaaa agaaccgggc gcccctgcgc tgacagccgg aacacggcgg catcagagca
     2761 gccgattgtc tgttgtgccc agtcatagcc gaatagcctc tccacccaag cggccggaga
     2821 acctgcgtgc aatccatctt gttcaatcat ggtggcatcg attcccggtc aacatccagt
     2881 acacctccgg cacaccggct tccccaaggg cgccacgctc tcccaccaca acatcctcaa
     2941 caacggctac tgggtgggcc gcacggtcgg ctacacggag caggaccggg tgtgtctgcc
     3001 ggtgcccttc taccactgct tcggcatggt catgggcaac ttgggcgcca cctcccacgc
     3061 cgcctgcatc gtcatcccgg ccccgtcctt cgagccggcg gcgacgctgg aggcggtgca
     3121 gcgggagagg tgcacgtccc tgtacggcgt ccccacgaag ccgaattcca ccacactgga
     3181 ctagatgcat gctcgacagt gtgatggata tctgcagaat tcggctttca cgacacctga
     3241 aatggaagaa aaaaactttg aaccactgtc tgaggcttga gaatgaacca agatccaaac
     3301 tcaaaaaggg caaattccaa ggagaattac atcaagtgcc aagctggcct aacttcagtc
     3361 tccacccact cagtgtgggg aaactccatc gcataaaacc cctcccccca acctaaagac
     3421 gacgtactcc aaaagctcga gaactaatcg aggtgcctgg acggcgcccg gtactccgtg
     3481 gagtcacatg aagcgacggc tgaggacgga aaggcccttt tcctttgtgt gggtgactca
     3541 cccgcccgct ctcccgagcg ccgcgtcctc cattttgagc tgcctgcagc agggccggga
     3601 agcggccatc tttccgctca cgcaactggt gccgaccggg ccagccttgc cgcccagggc
     3661 ggggcgatac acggcggcgc gaggccaggc accagagcag gccggccagc ttgagactac
     3721 ccccgtccga ttctcggtgg ccgcgctcgc aggccccgcc tcgccgaaca tgtgcgctgg
     3781 gacgcacggg ccccgtcgcc gcccgcggcc ccaaaaaccg aaataccagt gtgcagatct
     3841 tggcccgcat ttacaagact atcttgccag aaaaaaagcg tcgcagcagg tcatcaaaaa
     3901 ttttaaatgg ctagagactt atcgaaagca gcgagacagg cgcgaaggtg ccaccagatt
     3961 cgcacgcggc ggccccagcg cccaggccag gcctcaactc aagcacgagg cgaaggggct
     4021 ccttaagcgc aaggcctcga actctcccac ccacttccaa cccgaagctc gggatcaaga
     4081 atcacgtact gcagccaggt ggaagtaatt caaggcacgc aagggccata acccgtaaag
     4141 aggccaggcc cgcgggaacc acacacggca cttacctgtg ttctggcggc aaacccgttg
     4201 cgaaaaagaa cgttcacggc gactactgca cttatatacg gttctccccc accctcggga
     4261 aaaaggcgga gccagtacac gacatcactt tcccagttta ccccgcgcca ccttctctag
     4321 gcaccggttc aattgccgac ccctcccccc aacttctcgg ggactgtggg cgatgtgcgc
     4381 tctgcccact gacgggcacc ggagccaaga cgcgtgacgt cggcagtgaa aaaaatgctt
     4441 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     4501 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt
     4561 tttttaaagc aagtaaaacc tctacaaatg tggtatggct gattatgatc cgcggccgca
     4621 atacgtcgac gttatcagct gacttcgtac gagagcctag gtgtttctag agtcgcggcc
     4681 ttacgccaga atgcgttcgc acagccgcca gccggtcact ccgttgatgg ttactcggaa
     4741 cagcagggag ccgtcggggt tgatcaggcg ctcgtcgata attttgttgc cgttccacag
     4801 ggtccctgtt acagtgatct ttttgccgtc gaacacggcg atgccttcat acggccgtcc
     4861 gaaatagtcg atcatgttcg gcgtaacccc gtcgattacc agtgtgccat agtgcaggat
     4921 caccttaaag tgatgatcat ccacagggta caccacctta aaaatttttt cgatctggcc
     4981 catttggtcg ccgctcagac cttcatacgg gatgatgaca tggatgtcga tcttcagccc
     5041 attttcaccg ctcaggacaa tcctttggat cggagttacg gacaccccga gattctgaaa
     5101 caaactggac acacctccct gttcaaggac ttggtccagg ttgtagccgg ctgtctgtcg
     5161 ccagtcccca acgaaatctt cgagtgtgaa gacgccgctc gagccgagag aaactataat
     5221 atggaacctt ggtccaggtg ttgcgatgat gtcactgtac ggttcttcct gtgtcagctc
     5281 aatagcttgc tgctttttaa gaaccaagaa gctgtagaac tttgcggcag cttgttttct
     5341 gttcgtattt cgacataact caagcaaact gatagattca gctccagttt tagcaagagc
     5401 acgctgaaga ccatgaagca tctgctgagt ccttttgttc catcttcttt cttcctgatc
     5461 ttgatcgccc cctgatgcat cttcatcctc ttcctcttca tcatcttctt tttccttctc
     5521 tttctccttc tctttttctg gcagaagttc taactctggt attagctgac agatatttgg
     5581 aggttcttct gggggaagct ctacaggtgg tatttccatc tgctctacct gctgaggagg
     5641 catcacaggc tctgggtcaa tttgtccagc ttttcgctta actccctgag gtggtggtgg
     5701 aggcatagct gactcatcta tgtttgttct gctggcctcc atcactgact cctggaggcg
     5761 gcttggctct tcaataatgg gctcatcgat aacatcacgc tgctgatgct gctgttgctg
     5821 gtcctctcta ggaacctctg gattttcaaa ttctttgagg aattcatcca aattatctgc
     5881 ctctcctcct ttcctccttt ttctaaggtc ttctggtaca agcggtgtaa gacagcgtgt
     5941 aaagagcttc agtagtctgt tattccacaa aggctgagca ggtaaagaaa acagtttttc
     6001 tactcctcct gtctctttcc acatcatcaa tttcttggtg ggcggtgcca gatccaaagt
     6061 agtaacaata tctgaataat cactaagttg ggctctaatt gtcttgctat ccaactcttt
     6121 gacactgtca acaattagct tcctcttcct cttggctttt gtttctttaa cagttatatc
     6181 aataggctcc aatgcaaatg cttcttcctc atttggaaca agtgttgttt gatcagtcat
     6241 ggttggcatt ggttcaacgg gatccactga atcaggacta tcaggcccac ccattgatac
     6301 attatcatcc tcatccatat cgtcatgtgc aggctgctct ggcaacatca cccctgcctc
     6361 agagagggca gggggatcat caaagatacc gccatcatta ttactaataa gtttgtcatc
     6421 taatattcca ccatcatttc cttctccaaa attatcatcc ttatattgat cttcatattc
     6481 taaatggtta attttctcat tcagattgct ggtgctctgt tcagactcta ataggaggtt
     6541 agaagtagta gtgcttacta acatgtcgtc atcctcaaaa gcactgcctt ctctcattat
     6601 ctcacgatca tccattccaa aatcaccaaa atcattttct tgtaaaatac tgatgttccc
     6661 aacttcttct ctcatggtta tctcttccac tctactctga ttcaagctga actgctgggc
     6721 cacatcgatg tcatctaagt caggcagtgg ctgatcaaag tcatgaaatt cttcaggtaa
     6781 agtaatggca ttataagctg cttcccgatt ttcctcaggc aggtcaacca cacctggccg
     6841 aaaagccatc tttatcttaa tgaatgcttc attacagtct gcaagaaggt atttggcttt
     6901 cctgtgatag attcgaacta ctcccagtaa gagatgtcct gatgtccgta atgccatttt
     6961 cacctttggt gagatgatac tctccacgct gctctctaaa ttacactcga acacatgggc
     7021 tttggttagc ttcttatccc aatgggccgc tagccaaatt ttggccagag gccctctttt
     7081 actgagaaca aaatgtgcgt agaacatggc gatcgctagc tgcaaaaaga acaagtagct
     7141 tgtattctat agtgtcacct aaatgggcga tctgacggtt cactaaacca gctctgctta
     7201 tataggcctc cctggcgcgc cactagttat taatagtaat caattacggg gtcattagtt
     7261 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
     7321 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
     7381 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccact
//
