LOCUS       sequence-475188-        6333 bp    DNA     circular SYN 24-JUN-2025
DEFINITION  Express RBM39-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 6333)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 6333)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 24, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: RBM39-NanoLuc Vector
FEATURES             Location/Qualifiers
     source          1..6333
                     /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             2676..3188
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     primer_bind     complement(3276..3294)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3331..3452
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3342..3361)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3396..3415
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3697..4054
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3697..3888
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3720..3740)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3905..4040
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4069..4116
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4148..4942
                     /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(4202..4221)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4812..4831
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5006..5054
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(5293..5881)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5373..5392)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5997..6280
                     /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 ggctagcgat cgccatggca gacgatattg
     1081 atattgaagc aatgcttgag gctccttaca agaaggatga gaacaagttg agcagtgcca
     1141 acggccatga agaacgtagc aaaaagagga aaaaaagcaa gagcagaagt cgtagtcatg
     1201 aacgaaagag aagcaaaagt aaggaacgga agcgaagtag agacagagaa aggaaaaaga
     1261 gcaaaagccg tgaaagaaag cgaagtagaa gcaaagagag gcgacggagc cgctcaagaa
     1321 gtcgagatcg aagatttaga ggccgctaca gaagtcctta ctccggacca aaatttaaca
     1381 gtgccatccg aggaaagatt gggttgcctc atagcatcaa attaagcaga cgacgttccc
     1441 gaagcaaaag tccattcaga aaagacaaga gccctgtgag agaacctatt gataatttaa
     1501 ctcctgagga aagagatgca aggacagtct tctgtatgca gctggcggca agaattcgac
     1561 caagggattt ggaagagttt ttctctacag taggaaaggt tcgagatgtg aggatgattt
     1621 ctgacagaaa ttcaagacgt tccaaaggaa ttgcttatgt ggagttcgtc gatgttagct
     1681 cagtgcctct agcaatagga ttaactggcc aacgagtttt aggcgtgcca atcatagtac
     1741 aggcatcaca ggcagaaaaa aacagagctg cagcaatggc aaacaattta caaaagggaa
     1801 gtgctggacc tatgaggctt tatgtgggct cattacactt caacataact gaagatatgc
     1861 ttcgtgggat ctttgagcct tttggaagaa ttgaaagtat ccagctgatg atggacagtg
     1921 aaactggtcg atccaaggga tatggattta ttacattttc tgactcagaa tgtgccaaaa
     1981 aggctttgga acaacttaat ggatttgaac tagcaggaag accaatgaaa gttggtcatg
     2041 ttactgaacg tactgatgct tcgagtgcta gttcattttt ggacagtgat gaactggaaa
     2101 ggactggaat tgatttggga acaactggtc gtcttcagtt aatggcaaga cttgcagagg
     2161 gtacaggttt gcagattccg ccagcagcac agcaagctct acagatgagt ggctctttgg
     2221 catttggtgc tgtggcagaa ttctcttttg ttatagattt gcaaacaaga ctttcccagc
     2281 agactgaagc ttcagcttta gctgcagctg cctctgttca gccacttgca acacaatgtt
     2341 tccaactctc taacatgttt aaccctcaaa cagaagaaga agttggatgg gataccgaga
     2401 ttaaggatga tgtgattgaa gaatgtaata aacatggagg agttattcat atttatgttg
     2461 acaaaaattc agctcagggc aatgtgtatg tgaagtgccc atcaattgct gcagctattg
     2521 ctgctgtcaa tgcattgcat ggcaggtggt ttgctggtaa aatgataaca gcagcatatg
     2581 tacctcttcc aacttaccac aacctgtttc ctgattctat gacagcaaca cagctactgg
     2641 ttccaagtag acgagtttct ctcggctcga gcggcgtctt cacactcgaa gatttcgttg
     2701 gggactggcg acagacagcc ggctacaacc tggaccaagt ccttgaacag ggaggtgtgt
     2761 ccagtttgtt tcagaatctc ggggtgtccg taactccgat ccaaaggatt gtcctgagcg
     2821 gtgaaaatgg gctgaagatc gacatccatg tcatcatccc gtatgaaggt ctgagcggcg
     2881 accaaatggg ccagatcgaa aaaattttta aggtggtgta ccctgtggat gatcatcact
     2941 ttaaggtgat cctgcactat ggcacactgg taatcgacgg ggttacgccg aacatgatcg
     3001 actatttcgg acggccgtat gaaggcatcg ccgtgttcga cggcaaaaag atcactgtaa
     3061 cagggaccct gtggaacggc aacaaaatta tcgacgagcg cctgatcaac cccgacggct
     3121 ccctgctgtt ccgagtaacc atcaacggag tgaccggctg gcggctgtgc gaacgcattc
     3181 tggcgtaagg ccgcgactct agagtcgacc tgcaggcatg caagctgatc cggctgctaa
     3241 caaagcccga aaggaagctg agttggctgc tgccaccgct gagcaataac tagcataacc
     3301 ccttggggcg gccgcttcga gcagacatga taagatacat tgatgagttt ggacaaacca
     3361 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
     3421 ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt
     3481 ttcaggttca gggggagatg tgggaggttt ttttaagcaa gtaaaacctc tacaaatgtg
     3541 gtaaaatcga attttaacaa aatattaacg cttacaattt cctgatgcgg tattttctcc
     3601 ttacgcatct gtgcggtatt tcacaccgca tacgcggatc tgcgcagcac catggcctga
     3661 aataacctct gaaagaggaa cttggttagg taccttctga ggcggaaaga accagctgtg
     3721 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3781 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     3841 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     3901 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     3961 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     4021 aggaggcttt tttggaggcc taggcttttg caaaaagctt aattaactgt tgacaattaa
     4081 tcatcggcat agtatatcgg catagtataa tacgacaagg tgaggaacta aacccaggag
     4141 gcagatcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     4201 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     4261 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     4321 tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     4381 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     4441 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     4501 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     4561 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     4621 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat
     4681 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     4741 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     4801 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     4861 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     4921 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     4981 cccaacctgc catcacgatg gccgcaataa aatatcttta ttttcattac atctgtgtgt
     5041 tggttttttg tgtgaatcga tagcgataag gatcctcttt gcgcttgcgt tttcccttgt
     5101 ccagatagcc cagtagctga cattcatccg gggtcagcac cgtttctgcg gactggcttt
     5161 ctacccggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     5221 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     5281 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     5341 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     5401 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     5461 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     5521 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     5581 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     5641 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     5701 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     5761 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     5821 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatttca
     5881 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     5941 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt tatagtccgg
     6001 aaatacagga acgcacgctg gatggccctt cgctgggatg gtgaaaccat gaaaaatggc
     6061 agcttcagtg gattaagtgg gggtaatgtg gcctgtaccc tctggttgca taggtattca
     6121 tacggttaaa atttatcagg cgcgattgcg gcagtttttc gggtggtttg ttgccatttt
     6181 tacctgtctg ctgccgtgat cgcgctgaac gcgttttagc ggtgcgtaca attaagggat
     6241 tatggtaaat ccacttactg tctgccctcg tagccatcga gataaaccgc agtactccgg
     6301 ccacgatgcg tccggcgtag aggatcgaga tct
//
