LOCUS       sequence-471883-        7981 bp    DNA     circular SYN 14-JUL-2025
DEFINITION  Express NanoLuc(R)-EIF2AK3 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 7981)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 7981)
  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-EIF2AK3 Fusion Vector
FEATURES             Location/Qualifiers
     source          1..7981
                     /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(5060..5078)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    5115..5236
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5126..5145)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5180..5199
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        5481..5838
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5481..5672
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5504..5524)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5689..5824
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        5853..5900
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5932..6726
                     /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(5986..6005)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6596..6615
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6790..6838
                     /note="synthetic polyadenylation signal"
     rep_origin      6995..7540
                     /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     7441..7460
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7657..7674
                     /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 ggagcgcgcc atcagcccgg
     1621 ggctgctggt acgggcgctg ctgctgctgc tgctgctgct ggggctcgcg gcaaggacgg
     1681 tggccgcggg gcgcgcccgt ggcctcccag cgccgacggc ggaggcggcg ttcggcctcg
     1741 gggcggccgc tgctcccacc tcagcgacgc gagtaccggc ggcgggcgcc gtggctgcgg
     1801 ccgaggtgac tgtggaggac gctgaggcgc tgccggcagc cgcgggagag caggagcctc
     1861 ggggtccgga accagacgat gagacagagt tgcgaccgcg cggcaggtca ttagtaatta
     1921 tcagcacttt agatgggaga attgctgcct tggatcctga aaatcatggt aaaaagcagt
     1981 gggatttgga tgtgggatcc ggttccttgg tgtcatccag ccttagcaaa ccagaggtat
     2041 ttgggaataa gatgatcatt ccttccctgg atggagccct cttccagtgg gaccaagacc
     2101 gtgaaagcat ggaaacagtt cctttcacag ttgaatcact tcttgaatct tcttataaat
     2161 ttggagatga tgttgttttg gttggaggaa aatctctgac tacatatgga ctcagtgcat
     2221 atagtggaaa ggtgaggtat atctgttcag ctctgggttg tcgccaatgg gatagtgacg
     2281 aaatggaaca agaggaagac atcctgcttc tacagcgtac ccaaaaaact gttagagctg
     2341 tcggacctcg cagtggcaat gagaagtgga atttcagtgt tggccacttt gaacttcggt
     2401 atattccaga catggaaacg agagccggat ttattgaaag cacctttaag cccaatgaga
     2461 acacagaaga gtctaaaatt atttcagatg tggaagaaca ggaagctgcc ataatggaca
     2521 tagtgataaa ggtttcggtt gctgactgga aagttatggc attcagtaag aagggaggac
     2581 atctggaatg ggagtaccag ttttgtactc caattgcatc tgcctggtta cttaaggatg
     2641 ggaaagtcat tcccatcagt ctttttgatg atacaagtta tacatctaat gatgatgttt
     2701 tagaagatga agaagacatt gtagaagctg ccagaggagc cacagaaaac agtgtttact
     2761 tgggaatgta tagaggccag ctgtatctgc agtcatcagt cagaatttca gaaaagtttc
     2821 cttcaagtcc caaggctttg gaatctgtca ctaatgaaaa cgcaattatt cctttaccaa
     2881 caatcaaatg gaaaccctta attcattctc cttccagaac tcctgtcttg gtaggatctg
     2941 atgaatttga caaatgtctc agtaatgata agttttctca tgaagaatat agtaatggtg
     3001 cactttcaat cttgcagtat ccatatgata atggttatta tctaccatac tacaagaggg
     3061 agaggaacaa acgaagcaca cagattacag tcagattcct cgacaaccca cattacaaca
     3121 agaatatccg caaaaaggat cctgttcttc ttttacactg gtggaaagaa atagttgcaa
     3181 cgattttgtt ttgtatcata gcaacaacgt ttattgtgcg caggcttttc catcctcatc
     3241 ctcacaggca aaggaaggag tctgaaactc agtgtcaaac tgaaaataaa tatgattctg
     3301 taagtggtga agccaatgac agtagctgga atgacataaa aaactctgga tatatatcac
     3361 gatatctaac tgattttgag ccaattcaat gcctgggacg tggtggcttt ggagttgttt
     3421 ttgaagctaa aaacaaagta gatgactgca attatgctat caagaggatc cgtctcccca
     3481 atagggaatt ggctcgggaa aaggtaatgc gagaagttaa agccttagcc aagcttgaac
     3541 acccgggcat tgttagatat ttcaatgcct ggctcgaagc accaccagag aagtggcaag
     3601 aaaagatgga tgaaatttgg ctgaaagatg aaagcacaga ctggccactc agctctccta
     3661 gcccaatgga tgcaccatca gttaaaatac gcagaatgga tcctttcgct acaaaagaac
     3721 atattgaaat catagctcct tcaccacaaa gaagcaggtc tttttcagta gggatttcct
     3781 gtgaccagac aagttcatct gagagccagt tctcaccact ggaattctca ggaatggacc
     3841 atgaggacat cagtgagtca gtggatgcag catacaacct ccaggacagt tgccttacag
     3901 actgtgatgt ggaagatggg actatggatg gcaatgatga ggggcactcc tttgaacttt
     3961 gtccttctga agcttctcct tatgtaaggt caagggagag aacctcctct tcaatagtat
     4021 ttgaagattc tggctgtgat aatgcttcca gtaaagaaga gccgaaaact aatcgattgc
     4081 atattggcaa ccattgtgct aataaactaa ctgctttcaa gcccaccagt agcaaatctt
     4141 cttctgaagc tacattgtct atttctcctc caagaccaac cactttaagt ttagatctca
     4201 ctaaaaacac cacagaaaaa ctccagccca gttcaccaaa ggtgtatctt tacattcaaa
     4261 tgcagctgtg cagaaaagaa aacctcaaag actggatgaa tggacgatgt accatagagg
     4321 agagagagag gagcgtgtgt ctgcacatct tcctgcagat cgcagaggca gtggagtttc
     4381 ttcacagtaa aggactgatg cacagggacc tcaagccatc caacatattc tttacaatgg
     4441 atgatgtggt caaggttgga gactttgggt tagtgactgc aatggaccag gatgaggaag
     4501 agcagacggt tctgacccca atgccagctt atgccagaca cacaggacaa gtagggacca
     4561 aactgtatat gagcccagag cagattcatg gaaacagcta ttctcataaa gtggacatct
     4621 tttctttagg cctgattcta tttgaattgc tgtatccatt cagcactcag atggagagag
     4681 tcaggacctt aactgatgta agaaatctca aatttccacc attatttact cagaaatatc
     4741 cttgtgagta cgtgatggtt caagacatgc tctctccatc ccccatggaa cgacctgaag
     4801 ctataaacat cattgaaaat gctgtatttg aggacttgga ctttccagga aaaacagtgc
     4861 tcagacagag gtctcgctcc ttgagttcat cgggaacaaa acattcaaga cagtccaaca
     4921 actcccatag ccctttgcca agcaatgttt aaacgaattc gggctcggta cccggggatc
     4981 ctctagagtc gacctgcagg catgcaagct gatccggctg ctaacaaagc ccgaaaggaa
     5041 gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcggccgct
     5101 tcgagcagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     5161 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     5221 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga
     5281 gatgtgggag gtttttttaa gcaagtaaaa cctctacaaa tgtggtaaaa tcgaatttta
     5341 acaaaatatt aacgcttaca atttcctgat gcggtatttt ctccttacgc atctgtgcgg
     5401 tatttcacac cgcatacgcg gatctgcgca gcaccatggc ctgaaataac ctctgaaaga
     5461 ggaacttggt taggtacctt ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta
     5521 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     5581 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     5641 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     5701 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     5761 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     5821 ggcctaggct tttgcaaaaa gcttaattaa ctgttgacaa ttaatcatcg gcatagtata
     5881 tcggcatagt ataatacgac aaggtgagga actaaaccca ggaggcagat catgattgaa
     5941 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     6001 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     6061 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     6121 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     6181 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     6241 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     6301 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     6361 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     6421 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     6481 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     6541 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     6601 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     6661 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     6721 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     6781 gatggccgca ataaaatatc tttattttca ttacatctgt gtgttggttt tttgtgtgaa
     6841 tcgatagcga taaggatccg ctcatgcgat gataagctgt caaacatgag aattacaact
     6901 tatatcgtat ggggctgact tcaggtgcta catttgaaga gataaattgc actgaaatct
     6961 agaaatattt tatctgatta ataagatgat cttcttgaga tcgttttggt ctgcgcgtaa
     7021 tctcttgctc tgaaaacgaa aaaaccgcct tgcagggcgg tttttcgaag gttctctgag
     7081 ctaccaactc tttgaaccga ggtaactggc ttggaggagc gcagtcacca aaacttgtcc
     7141 tttcagttta gccttaaccg gcgcatgact tcaagactaa ctcctctaaa tcaattacca
     7201 gtggctgctg ccagtggtgc ttttgcatgt ctttccgggt tggactcaag acgatagtta
     7261 ccggataagg cgcagcggtc ggactgaacg gggggttcgt gcatacagtc cagcttggag
     7321 cgaactgcct acccggaact gagtgtcagg cgtggaatga gacaaacgcg gccataacag
     7381 cggaatgaca ccggtaaacc gaaaggcagg aacaggagag cgcacgaggg agccgccagg
     7441 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc caccactgat ttgagcgtca
     7501 gatttcgtga tgcttgtcag gggggcggag cctatggaaa aacggctttg ccgcggccct
     7561 ctcacttccc tgttaagtat cttcctggca tcttccagga aatctccgcc ccgttcgtaa
     7621 gccatttccg ctcgccgcag tcgaacgacc gagcgtagcg agtcagtgag cgaggaagcg
     7681 gaatatatcc tgtatcacat attctgctga cgcaccggtg cagccttttt tctcctgcca
     7741 catgaagcac ttcactgaca ccctcatcag tgccaacata gtaagccagt atacactccg
     7801 ctagcgctga tgtccggcgg tgcttttgcc gttacgcacc accccgtcat gagattacgc
     7861 gttttagcgg tgcgtacaat taagggatta tggtaaatcc acttactgtc tgccctcgta
     7921 gccatcgaga taaaccgcag tactccggcc acgatgcgtc cggcgtagag gatcgagatc
     7981 t
//
