LOCUS       sequence-473336-        5282 bp    DNA     circular SYN 14-JUL-2025
DEFINITION  Express NanoLuc(R)-MBP 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 5282)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 5282)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: NanoLuc-MBP Fusion Vector
FEATURES             Location/Qualifiers
     source          1..5282
                     /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"
     primer_bind     complement(2225..2243)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2280..2401
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2291..2310)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2345..2364
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2646..3003
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2646..2837
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2669..2689)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2854..2989
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3018..3065
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3097..3891
                     /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(3151..3170)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3761..3780
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3955..4003
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(4242..4830)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4322..4341)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    4946..5229
                     /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 ggcgtcacag aagagaccct
     1621 cccagaggca cggatccaag tacctggcca cagcaagtac catggaccat gccaggcatg
     1681 gcttcctccc aaggcacaga gacacgggca tccttgactc catcgggcgc ttctttggcg
     1741 gtgacagggg tgcgcccaag cggggctctg gcaaggactc acaccacccg gcaagaactg
     1801 ctcactacgg ctccctgccc cagaagtcac acggccggac ccaagatgaa aaccccgtag
     1861 tccacttctt caagaacatt gtgacgcctc gcacaccacc cccgtcgcag ggaaagggga
     1921 gaggactgtc cctgagcaga tttagctggg gggccgaagg ccagagacca ggatttggct
     1981 acggaggcag agcgtccgac tataaatcgg ctcacaaggg attcaaggga gtcgatgccc
     2041 agggcacgct ttccaaaatt tttaagctgg gaggaagaga tagtcgctct ggatcaccca
     2101 tggctagacg cgtttaaacg aattcgggct cggtacccgg ggatcctcta gagtcgacct
     2161 gcaggcatgc aagctgatcc ggctgctaac aaagcccgaa aggaagctga gttggctgct
     2221 gccaccgctg agcaataact agcataaccc cttggggcgg ccgcttcgag cagacatgat
     2281 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     2341 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     2401 taacaacaac aattgcattc attttatgtt tcaggttcag ggggagatgt gggaggtttt
     2461 tttaagcaag taaaacctct acaaatgtgg taaaatcgaa ttttaacaaa atattaacgc
     2521 ttacaatttc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     2581 acgcggatct gcgcagcacc atggcctgaa ataacctctg aaagaggaac ttggttaggt
     2641 accttctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     2701 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     2761 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     2821 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     2881 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     2941 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     3001 aaaaagctta attaactgtt gacaattaat catcggcata gtatatcggc atagtataat
     3061 acgacaaggt gaggaactaa acccaggagg cagatcatga ttgaacaaga tggattgcac
     3121 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     3181 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     3241 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     3301 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     3361 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     3421 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     3481 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     3541 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     3601 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     3661 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     3721 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     3781 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     3841 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     3901 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgatgg ccgcaataaa
     3961 atatctttat tttcattaca tctgtgtgtt ggttttttgt gtgaatcgat agcgataagg
     4021 atcctctttg cgcttgcgtt ttcccttgtc cagatagccc agtagctgac attcatccgg
     4081 ggtcagcacc gtttctgcgg actggctttc tacccggtat cagctcactc aaaggcggta
     4141 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     4201 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     4261 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     4321 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     4381 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     4441 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     4501 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     4561 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     4621 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     4681 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     4741 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     4801 cagattacgc gcagaaaaaa aggatttcaa gaagatcctt tgatcttttc tacggggtct
     4861 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     4921 atcttcacct agatcctttt atagtccgga aatacaggaa cgcacgctgg atggcccttc
     4981 gctgggatgg tgaaaccatg aaaaatggca gcttcagtgg attaagtggg ggtaatgtgg
     5041 cctgtaccct ctggttgcat aggtattcat acggttaaaa tttatcaggc gcgattgcgg
     5101 cagtttttcg ggtggtttgt tgccattttt acctgtctgc tgccgtgatc gcgctgaacg
     5161 cgttttagcg gtgcgtacaa ttaagggatt atggtaaatc cacttactgt ctgccctcgt
     5221 agccatcgag ataaaccgca gtactccggc cacgatgcgt ccggcgtaga ggatcgagat
     5281 ct
//
