LOCUS       sequence-479792-        7564 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Express NanoLuc-ERN1 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 7564)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7564)
  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            "Unpublished"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-ERN1 Fusion Vector
FEATURES             Location/Qualifiers
     source          1..7564
                     /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(4643..4661)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    4698..4819
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4709..4728)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4763..4782
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        5064..5421
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5064..5255
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5087..5107)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5272..5407
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        5436..5483
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5515..6309
                     /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(5569..5588)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6179..6198
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6373..6421
                     /note="synthetic polyadenylation signal"
     rep_origin      6578..7123
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     7024..7043
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7240..7257
                     /label=L4440
                     /note="L4440 vector, forward primer"
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 gccggcccgg cggctgctgc
     1621 tgctgctgac gctgctgctg cccggcctcg ggatttttgg aagtaccagc acagtgacgc
     1681 ttcctgaaac cttgttgttt gtgtcaacgc tggatggaag tttgcatgct gtcagcaaga
     1741 ggacaggctc aatcaaatgg actttaaaag aagatccagt cctgcaggtc ccaacacatg
     1801 tggaagagcc tgcctttctc ccagatccta atgatggcag cctgtatacg cttggaagca
     1861 agaataatga aggcctgacg aaacttcctt ttaccatccc agaattggtg caggcatccc
     1921 catgccgaag ttcagatgga atcctctaca tgggtaaaaa gcaggacatc tggtatgtta
     1981 ttgacctcct gaccggagag aagcagcaga ctttgtcatc ggcctttgca gatagtctct
     2041 gcccatcaac ctctcttctg tatcttgggc gaacagaata caccatcacc atgtacgaca
     2101 ccaaaacccg agagctccgg tggaatgcca cctactttga ctatgcggcc tcactgcctg
     2161 aggacgacgt ggactacaag atgtcccact ttgtgtccaa tggtgatggg ctggtggtga
     2221 ctgtggacag tgaatctggg gacgtcctgt ggatccaaaa ctacgcctcc cctgtggtgg
     2281 ccttttatgt ctggcagcgg gagggtctga ggaaggtgat gcacatcaat gtcgctgtgg
     2341 agaccctgcg ctatctgacc ttcatgtctg gggaggtggg gcgcatcaca aagtggaagt
     2401 acccgttccc caaggagaca gaggccaaga gcaagctgac gcccactctg tatgttggga
     2461 aatactctac cagcctctat gcctctccct caatggtaca cgagggggtt gctgtcgtgc
     2521 cccgcggcag cacacttcct ttgctggaag ggccccagac tgatggcgtc accatcgggg
     2581 acaaggggga gtgtgtgatc acgcccagca cggacgtcaa gtttgatccc ggactcaaaa
     2641 gcaagaacaa gctcaactac ttgaggaatt actggcttct gataggacac catgaaaccc
     2701 cactgtctgc gtctaccaag atgctggaga gatttcccaa caatctaccc aaacatcggg
     2761 aaaatgtgat tcctgctgat tcagagaaaa agagctttga ggaagttatc aacctggttg
     2821 accagacttc agaaaacgca cctaccaccg tgtctcggga tgtggaggag aagcccgccc
     2881 atgcccctgc ccggcccgag gcccccgtgg actccatgct taaggacatg gctaccatca
     2941 tcctgagcac cttcctgctg attggctggg tggccttcat catcacctat cccctgagca
     3001 tgcatcagca gcagcagctc cagcaccagc agttccagaa ggaactggag aagatccagc
     3061 tcctgcagca gcagcagcag cagctgccct tccacccacc tggagacacg gctcaggacg
     3121 gcgagctcct ggacacgtct ggcccgtact cagagagctc gggcaccagc agccccagca
     3181 cgtcccccag ggcctccaac cactcgctct gctccggcag ctctgcctcc aaggctggca
     3241 gcagcccctc cctggaacaa gacgatggag atgaggaaac cagcgtggtg atagttggga
     3301 aaatttcctt ctgtcccaag gatgtcctgg gccatggagc tgagggcaca attgtgtacc
     3361 ggggcatgtt tgacaaccgc gacgtggccg tgaagaggat cctccccgag tgttttagct
     3421 tcgcagaccg tgaggtccag ctgttgcgag aatcggatga gcacccgaac gtgatccgct
     3481 acttctgcac ggagaaggac cggcaattcc agtacattgc catcgaactg tgtgcagcca
     3541 ccctgcaaga gtatgtggag cagaaggact ttgcgcatct cggcctggag cccatcacct
     3601 tgctgcagca gaccacctcg ggcctggccc acctccactc cctcaacatc gttcacagag
     3661 acctaaagcc acacaacatc ctcatatcca tgcccaatgc acacggcaag atcaaggcca
     3721 tgatctccga ctttggcctc tgcaagaagc tggcagtggg cagacacagt ttcagccgcc
     3781 gatctggggt gcctggcaca gaaggctgga tcgctccaga gatgctgagc gaagactgta
     3841 aggagaaccc tacctacacg gtggacatct tttctgcagg ctgcgtcttt tactacgtaa
     3901 tctctgaggg cagccaccct tttggcaagt ccctgcagcg gcaggccaac atcctcctgg
     3961 gtgcctgcag ccttgactgc ttgcacccag agaagcacga agacgtcatt gcacgtgaat
     4021 tgatagagaa gatgattgcg atggatcctc agaaacgccc ctcagcgaag cacgtgctca
     4081 aacacccgtt cttctggagc ctagagaagc agctccagtt cttccaggac gtgagcgaca
     4141 gaatagaaaa ggaatccctg gatggcccga tcgtgaagca gttagagaga ggcgggagag
     4201 ccgtggtgaa gatggactgg cgggagaaca tcactgtccc cctccagaca gacctgcgta
     4261 aattcaggac ctataaaggt ggttctgtca gagatctcct ccgagccatg agaaataaga
     4321 agcaccacta ccgggagctg cctgcagagg tgcgggagac gctggggtcc ctccccgacg
     4381 acttcgtgtg ctacttcacg tctcgcttcc cccacctcct cgcacacacc taccgggcca
     4441 tggagctgtg cagccacgag agactcttcc agccctacta cttccacgag cccccagagc
     4501 cccagccccc agtgactcca gacgccctcg tttaaacgaa ttcgggctcg gtacccgggg
     4561 atcctctaga gtcgacctgc aggcatgcaa gctgatccgg ctgctaacaa agcccgaaag
     4621 gaagctgagt tggctgctgc caccgctgag caataactag cataacccct tggggcggcc
     4681 gcttcgagca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca
     4741 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
     4801 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg
     4861 ggagatgtgg gaggtttttt taagcaagta aaacctctac aaatgtggta aaatcgaatt
     4921 ttaacaaaat attaacgctt acaatttcct gatgcggtat tttctcctta cgcatctgtg
     4981 cggtatttca caccgcatac gcggatctgc gcagcaccat ggcctgaaat aacctctgaa
     5041 agaggaactt ggttaggtac cttctgaggc ggaaagaacc agctgtggaa tgtgtgtcag
     5101 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     5161 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     5221 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     5281 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     5341 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     5401 ggaggcctag gcttttgcaa aaagcttaat taactgttga caattaatca tcggcatagt
     5461 atatcggcat agtataatac gacaaggtga ggaactaaac ccaggaggca gatcatgatt
     5521 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     5581 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     5641 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     5701 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     5761 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     5821 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     5881 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     5941 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     6001 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     6061 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     6121 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     6181 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     6241 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     6301 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     6361 cacgatggcc gcaataaaat atctttattt tcattacatc tgtgtgttgg ttttttgtgt
     6421 gaatcgatag cgataaggat ccgctcatgc gatgataagc tgtcaaacat gagaattaca
     6481 acttatatcg tatggggctg acttcaggtg ctacatttga agagataaat tgcactgaaa
     6541 tctagaaata ttttatctga ttaataagat gatcttcttg agatcgtttt ggtctgcgcg
     6601 taatctcttg ctctgaaaac gaaaaaaccg ccttgcaggg cggtttttcg aaggttctct
     6661 gagctaccaa ctctttgaac cgaggtaact ggcttggagg agcgcagtca ccaaaacttg
     6721 tcctttcagt ttagccttaa ccggcgcatg acttcaagac taactcctct aaatcaatta
     6781 ccagtggctg ctgccagtgg tgcttttgca tgtctttccg ggttggactc aagacgatag
     6841 ttaccggata aggcgcagcg gtcggactga acggggggtt cgtgcataca gtccagcttg
     6901 gagcgaactg cctacccgga actgagtgtc aggcgtggaa tgagacaaac gcggccataa
     6961 cagcggaatg acaccggtaa accgaaaggc aggaacagga gagcgcacga gggagccgcc
     7021 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccaccact gatttgagcg
     7081 tcagatttcg tgatgcttgt caggggggcg gagcctatgg aaaaacggct ttgccgcggc
     7141 cctctcactt ccctgttaag tatcttcctg gcatcttcca ggaaatctcc gccccgttcg
     7201 taagccattt ccgctcgccg cagtcgaacg accgagcgta gcgagtcagt gagcgaggaa
     7261 gcggaatata tcctgtatca catattctgc tgacgcaccg gtgcagcctt ttttctcctg
     7321 ccacatgaag cacttcactg acaccctcat cagtgccaac atagtaagcc agtatacact
     7381 ccgctagcgc tgatgtccgg cggtgctttt gccgttacgc accaccccgt catgagatta
     7441 cgcgttttag cggtgcgtac aattaaggga ttatggtaaa tccacttact gtctgccctc
     7501 gtagccatcg agataaaccg cagtactccg gccacgatgc gtccggcgta gaggatcgag
     7561 atct
//
