LOCUS       sequence-476615-        7955 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express EIF2AK3-NanoLuc(R) 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 7955)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7955)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: EIF2AK3-NanoLuc Fusion Vector
FEATURES             Location/Qualifiers
     source          1..7955
                     /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             4434..4946
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     primer_bind     complement(5034..5052)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    5089..5210
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5100..5119)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5154..5173
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        5455..5812
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5455..5646
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5478..5498)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5663..5798
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        5827..5874
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5906..6700
                     /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(5960..5979)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6570..6589
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6764..6812
                     /note="synthetic polyadenylation signal"
     rep_origin      6969..7514
                     /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     7415..7434
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7631..7648
                     /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 ggctagcgat cgccatggag cgcgccatca
     1081 gcccggggct gctggtacgg gcgctgctgc tgctgctgct gctgctgggg ctcgcggcaa
     1141 ggacggtggc cgcggggcgc gcccgtggcc tcccagcgcc gacggcggag gcggcgttcg
     1201 gcctcggggc ggccgctgct cccacctcag cgacgcgagt accggcggcg ggcgccgtgg
     1261 ctgcggccga ggtgactgtg gaggacgctg aggcgctgcc ggcagccgcg ggagagcagg
     1321 agcctcgggg tccggaacca gacgatgaga cagagttgcg accgcgcggc aggtcattag
     1381 taattatcag cactttagat gggagaattg ctgccttgga tcctgaaaat catggtaaaa
     1441 agcagtggga tttggatgtg ggatccggtt ccttggtgtc atccagcctt agcaaaccag
     1501 aggtatttgg gaataagatg atcattcctt ccctggatgg agccctcttc cagtgggacc
     1561 aagaccgtga aagcatggaa acagttcctt tcacagttga atcacttctt gaatcttctt
     1621 ataaatttgg agatgatgtt gttttggttg gaggaaaatc tctgactaca tatggactca
     1681 gtgcatatag tggaaaggtg aggtatatct gttcagctct gggttgtcgc caatgggata
     1741 gtgacgaaat ggaacaagag gaagacatcc tgcttctaca gcgtacccaa aaaactgtta
     1801 gagctgtcgg acctcgcagt ggcaatgaga agtggaattt cagtgttggc cactttgaac
     1861 ttcggtatat tccagacatg gaaacgagag ccggatttat tgaaagcacc tttaagccca
     1921 atgagaacac agaagagtct aaaattattt cagatgtgga agaacaggaa gctgccataa
     1981 tggacatagt gataaaggtt tcggttgctg actggaaagt tatggcattc agtaagaagg
     2041 gaggacatct ggaatgggag taccagtttt gtactccaat tgcatctgcc tggttactta
     2101 aggatgggaa agtcattccc atcagtcttt ttgatgatac aagttataca tctaatgatg
     2161 atgttttaga agatgaagaa gacattgtag aagctgccag aggagccaca gaaaacagtg
     2221 tttacttggg aatgtataga ggccagctgt atctgcagtc atcagtcaga atttcagaaa
     2281 agtttccttc aagtcccaag gctttggaat ctgtcactaa tgaaaacgca attattcctt
     2341 taccaacaat caaatggaaa cccttaattc attctccttc cagaactcct gtcttggtag
     2401 gatctgatga atttgacaaa tgtctcagta atgataagtt ttctcatgaa gaatatagta
     2461 atggtgcact ttcaatcttg cagtatccat atgataatgg ttattatcta ccatactaca
     2521 agagggagag gaacaaacga agcacacaga ttacagtcag attcctcgac aacccacatt
     2581 acaacaagaa tatccgcaaa aaggatcctg ttcttctttt acactggtgg aaagaaatag
     2641 ttgcaacgat tttgttttgt atcatagcaa caacgtttat tgtgcgcagg cttttccatc
     2701 ctcatcctca caggcaaagg aaggagtctg aaactcagtg tcaaactgaa aataaatatg
     2761 attctgtaag tggtgaagcc aatgacagta gctggaatga cataaaaaac tctggatata
     2821 tatcacgata tctaactgat tttgagccaa ttcaatgcct gggacgtggt ggctttggag
     2881 ttgtttttga agctaaaaac aaagtagatg actgcaatta tgctatcaag aggatccgtc
     2941 tccccaatag ggaattggct cgggaaaagg taatgcgaga agttaaagcc ttagccaagc
     3001 ttgaacaccc gggcattgtt agatatttca atgcctggct cgaagcacca ccagagaagt
     3061 ggcaagaaaa gatggatgaa atttggctga aagatgaaag cacagactgg ccactcagct
     3121 ctcctagccc aatggatgca ccatcagtta aaatacgcag aatggatcct ttcgctacaa
     3181 aagaacatat tgaaatcata gctccttcac cacaaagaag caggtctttt tcagtaggga
     3241 tttcctgtga ccagacaagt tcatctgaga gccagttctc accactggaa ttctcaggaa
     3301 tggaccatga ggacatcagt gagtcagtgg atgcagcata caacctccag gacagttgcc
     3361 ttacagactg tgatgtggaa gatgggacta tggatggcaa tgatgagggg cactcctttg
     3421 aactttgtcc ttctgaagct tctccttatg taaggtcaag ggagagaacc tcctcttcaa
     3481 tagtatttga agattctggc tgtgataatg cttccagtaa agaagagccg aaaactaatc
     3541 gattgcatat tggcaaccat tgtgctaata aactaactgc tttcaagccc accagtagca
     3601 aatcttcttc tgaagctaca ttgtctattt ctcctccaag accaaccact ttaagtttag
     3661 atctcactaa aaacaccaca gaaaaactcc agcccagttc accaaaggtg tatctttaca
     3721 ttcaaatgca gctgtgcaga aaagaaaacc tcaaagactg gatgaatgga cgatgtacca
     3781 tagaggagag agagaggagc gtgtgtctgc acatcttcct gcagatcgca gaggcagtgg
     3841 agtttcttca cagtaaagga ctgatgcaca gggacctcaa gccatccaac atattcttta
     3901 caatggatga tgtggtcaag gttggagact ttgggttagt gactgcaatg gaccaggatg
     3961 aggaagagca gacggttctg accccaatgc cagcttatgc cagacacaca ggacaagtag
     4021 ggaccaaact gtatatgagc ccagagcaga ttcatggaaa cagctattct cataaagtgg
     4081 acatcttttc tttaggcctg attctatttg aattgctgta tccattcagc actcagatgg
     4141 agagagtcag gaccttaact gatgtaagaa atctcaaatt tccaccatta tttactcaga
     4201 aatatccttg tgagtacgtg atggttcaag acatgctctc tccatccccc atggaacgac
     4261 ctgaagctat aaacatcatt gaaaatgctg tatttgagga cttggacttt ccaggaaaaa
     4321 cagtgctcag acagaggtct cgctccttga gttcatcggg aacaaaacat tcaagacagt
     4381 ccaacaactc ccatagccct ttgccaagca atgtttctct cggctcgagc ggcgtcttca
     4441 cactcgaaga tttcgttggg gactggcgac agacagccgg ctacaacctg gaccaagtcc
     4501 ttgaacaggg aggtgtgtcc agtttgtttc agaatctcgg ggtgtccgta actccgatcc
     4561 aaaggattgt cctgagcggt gaaaatgggc tgaagatcga catccatgtc atcatcccgt
     4621 atgaaggtct gagcggcgac caaatgggcc agatcgaaaa aatttttaag gtggtgtacc
     4681 ctgtggatga tcatcacttt aaggtgatcc tgcactatgg cacactggta atcgacgggg
     4741 ttacgccgaa catgatcgac tatttcggac ggccgtatga aggcatcgcc gtgttcgacg
     4801 gcaaaaagat cactgtaaca gggaccctgt ggaacggcaa caaaattatc gacgagcgcc
     4861 tgatcaaccc cgacggctcc ctgctgttcc gagtaaccat caacggagtg accggctggc
     4921 ggctgtgcga acgcattctg gcgtaaggcc gcgactctag agtcgacctg caggcatgca
     4981 agctgatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga
     5041 gcaataacta gcataacccc ttggggcggc cgcttcgagc agacatgata agatacattg
     5101 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
     5161 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
     5221 attgcattca ttttatgttt caggttcagg gggagatgtg ggaggttttt ttaagcaagt
     5281 aaaacctcta caaatgtggt aaaatcgaat tttaacaaaa tattaacgct tacaatttcc
     5341 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata cgcggatctg
     5401 cgcagcacca tggcctgaaa taacctctga aagaggaact tggttaggta ccttctgagg
     5461 cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     5521 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     5581 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     5641 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     5701 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     5761 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagcttaa
     5821 ttaactgttg acaattaatc atcggcatag tatatcggca tagtataata cgacaaggtg
     5881 aggaactaaa cccaggaggc agatcatgat tgaacaagat ggattgcacg caggttctcc
     5941 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     6001 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     6061 cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac
     6121 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     6181 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     6241 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     6301 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     6361 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     6421 caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     6481 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     6541 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     6601 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     6661 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
     6721 atgaccgacc aagcgacgcc caacctgcca tcacgatggc cgcaataaaa tatctttatt
     6781 ttcattacat ctgtgtgttg gttttttgtg tgaatcgata gcgataagga tccgctcatg
     6841 cgatgataag ctgtcaaaca tgagaattac aacttatatc gtatggggct gacttcaggt
     6901 gctacatttg aagagataaa ttgcactgaa atctagaaat attttatctg attaataaga
     6961 tgatcttctt gagatcgttt tggtctgcgc gtaatctctt gctctgaaaa cgaaaaaacc
     7021 gccttgcagg gcggtttttc gaaggttctc tgagctacca actctttgaa ccgaggtaac
     7081 tggcttggag gagcgcagtc accaaaactt gtcctttcag tttagcctta accggcgcat
     7141 gacttcaaga ctaactcctc taaatcaatt accagtggct gctgccagtg gtgcttttgc
     7201 atgtctttcc gggttggact caagacgata gttaccggat aaggcgcagc ggtcggactg
     7261 aacggggggt tcgtgcatac agtccagctt ggagcgaact gcctacccgg aactgagtgt
     7321 caggcgtgga atgagacaaa cgcggccata acagcggaat gacaccggta aaccgaaagg
     7381 caggaacagg agagcgcacg agggagccgc cagggggaaa cgcctggtat ctttatagtc
     7441 ctgtcgggtt tcgccaccac tgatttgagc gtcagatttc gtgatgcttg tcaggggggc
     7501 ggagcctatg gaaaaacggc tttgccgcgg ccctctcact tccctgttaa gtatcttcct
     7561 ggcatcttcc aggaaatctc cgccccgttc gtaagccatt tccgctcgcc gcagtcgaac
     7621 gaccgagcgt agcgagtcag tgagcgagga agcggaatat atcctgtatc acatattctg
     7681 ctgacgcacc ggtgcagcct tttttctcct gccacatgaa gcacttcact gacaccctca
     7741 tcagtgccaa catagtaagc cagtatacac tccgctagcg ctgatgtccg gcggtgcttt
     7801 tgccgttacg caccaccccg tcatgagatt acgcgtttta gcggtgcgta caattaaggg
     7861 attatggtaa atccacttac tgtctgccct cgtagccatc gagataaacc gcagtactcc
     7921 ggccacgatg cgtccggcgt agaggatcga gatct
//
