LOCUS       sequence-476789-        5507 bp    DNA     circular SYN 20-JUN-2025
DEFINITION  Express EIF2AK2(252-551)-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 5507)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 5507)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 20, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: EIF2AK2(252-551)-NanoLuc Fusion Vector
FEATURES             Location/Qualifiers
     source          1..5507
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     104..124
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          289..421
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     404..423
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     465..484
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        465..483
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1418..1930
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     primer_bind     complement(2018..2036)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2073..2194
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2084..2103)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2138..2157
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2439..2796
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2439..2630
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2462..2482)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2647..2782
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        2811..2858
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2890..3684
                     /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(2944..2963)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3554..3573
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3748..3796
                     /note="synthetic polyadenylation signal"
     rep_origin      3953..4498
                     /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     4399..4418
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4615..4632
                     /label=L4440
                     /note="L4440 vector, forward primer"
     enhancer        5077..5456
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(5457..5507,1..153)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
ORIGIN
        1 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
       61 caacgggact ttccaaaatg tcgtaataac cccgccccgt tgacgcaaat gggcggtagg
      121 cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca gatcactaga
      181 agctttattg cggtagttta tcacagttaa attgctaacg cagtcagtgc ttctgacaca
      241 acagtctcga acttaagctg cagaagttgg tcgtgaggca ctgggcaggt aagtatcaag
      301 gttacaagac aggtttaagg agaccaatag aaactgggct tgtcgagaca gagaagactc
      361 ttgcgtttct gataggcacc tattggtctt actgacatcc actttgcctt tctctccaca
      421 ggtgtccact cccagttcaa ttacagctct taaggctaga gtattaatac gactcactat
      481 agggctagcg atcgccatga aagaaacaaa gtatactgtg gacaagaggt ttggcatgga
      541 ttttaaagaa atagaattaa ttggctcagg tggatttggc caagttttca aagcaaaaca
      601 cagaattgac ggaaagactt acgttattaa acgtgttaaa tataataacg agaaggcgga
      661 gcgtgaagta aaagcattgg caaaacttga tcatgtaaat attgttcact acaatggctg
      721 ttgggatgga tttgattatg atcctgagac cagtgatgat tctcttgaga gcagtgatta
      781 tgatcctgag aacagcaaaa atagttcaag gtcaaagact aagtgccttt tcatccaaat
      841 ggaattctgt gataaaggga ccttggaaca atggattgaa aaaagaagag gcgagaaact
      901 agacaaagtt ttggctttgg aactctttga acaaataaca aaaggggtgg attatataca
      961 ttcaaaaaaa ttaattcata gagatcttaa gccaagtaat atattcttag tagatacaaa
     1021 acaagtaaag attggagact ttggacttgt aacatctctg aaaaatgatg gaaagcgaac
     1081 aaggagtaag ggaactttgc gatacatgag cccagaacag atttcttcgc aagactatgg
     1141 aaaggaagtg gacctctacg ctttggggct aattcttgct gaacttcttc atgtatgtga
     1201 cactgctttt gaaacatcaa agtttttcac agacctacgg gatggcatca tctcagatat
     1261 atttgataaa aaagaaaaaa ctcttctaca gaaattactc tcaaagaaac ctgaggatcg
     1321 acctaacaca tctgaaatac taaggacctt gactgtgtgg aagaaaagcc cagagaaaaa
     1381 tgaacgacac acatgtgttt ctctcggctc gagcggcgtc ttcacactcg aagatttcgt
     1441 tggggactgg cgacagacag ccggctacaa cctggaccaa gtccttgaac agggaggtgt
     1501 gtccagtttg tttcagaatc tcggggtgtc cgtaactccg atccaaagga ttgtcctgag
     1561 cggtgaaaat gggctgaaga tcgacatcca tgtcatcatc ccgtatgaag gtctgagcgg
     1621 cgaccaaatg ggccagatcg aaaaaatttt taaggtggtg taccctgtgg atgatcatca
     1681 ctttaaggtg atcctgcact atggcacact ggtaatcgac ggggttacgc cgaacatgat
     1741 cgactatttc ggacggccgt atgaaggcat cgccgtgttc gacggcaaaa agatcactgt
     1801 aacagggacc ctgtggaacg gcaacaaaat tatcgacgag cgcctgatca accccgacgg
     1861 ctccctgctg ttccgagtaa ccatcaacgg agtgaccggc tggcggctgt gcgaacgcat
     1921 tctggcgtaa ggccgcgact ctagagtcga cctgcaggca tgcaagctga tccggctgct
     1981 aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa
     2041 ccccttgggg cggccgcttc gagcagacat gataagatac attgatgagt ttggacaaac
     2101 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     2161 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     2221 gtttcaggtt cagggggaga tgtgggaggt ttttttaagc aagtaaaacc tctacaaatg
     2281 tggtaaaatc gaattttaac aaaatattaa cgcttacaat ttcctgatgc ggtattttct
     2341 ccttacgcat ctgtgcggta tttcacaccg catacgcgga tctgcgcagc accatggcct
     2401 gaaataacct ctgaaagagg aacttggtta ggtaccttct gaggcggaaa gaaccagctg
     2461 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     2521 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     2581 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     2641 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     2701 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     2761 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc ttaattaact gttgacaatt
     2821 aatcatcggc atagtatatc ggcatagtat aatacgacaa ggtgaggaac taaacccagg
     2881 aggcagatca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     2941 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     3001 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     3061 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     3121 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     3181 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     3241 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     3301 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     3361 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     3421 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     3481 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     3541 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     3601 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     3661 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     3721 cgcccaacct gccatcacga tggccgcaat aaaatatctt tattttcatt acatctgtgt
     3781 gttggttttt tgtgtgaatc gatagcgata aggatccgct catgcgatga taagctgtca
     3841 aacatgagaa ttacaactta tatcgtatgg ggctgacttc aggtgctaca tttgaagaga
     3901 taaattgcac tgaaatctag aaatatttta tctgattaat aagatgatct tcttgagatc
     3961 gttttggtct gcgcgtaatc tcttgctctg aaaacgaaaa aaccgccttg cagggcggtt
     4021 tttcgaaggt tctctgagct accaactctt tgaaccgagg taactggctt ggaggagcgc
     4081 agtcaccaaa acttgtcctt tcagtttagc cttaaccggc gcatgacttc aagactaact
     4141 cctctaaatc aattaccagt ggctgctgcc agtggtgctt ttgcatgtct ttccgggttg
     4201 gactcaagac gatagttacc ggataaggcg cagcggtcgg actgaacggg gggttcgtgc
     4261 atacagtcca gcttggagcg aactgcctac ccggaactga gtgtcaggcg tggaatgaga
     4321 caaacgcggc cataacagcg gaatgacacc ggtaaaccga aaggcaggaa caggagagcg
     4381 cacgagggag ccgccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     4441 ccactgattt gagcgtcaga tttcgtgatg cttgtcaggg gggcggagcc tatggaaaaa
     4501 cggctttgcc gcggccctct cacttccctg ttaagtatct tcctggcatc ttccaggaaa
     4561 tctccgcccc gttcgtaagc catttccgct cgccgcagtc gaacgaccga gcgtagcgag
     4621 tcagtgagcg aggaagcgga atatatcctg tatcacatat tctgctgacg caccggtgca
     4681 gccttttttc tcctgccaca tgaagcactt cactgacacc ctcatcagtg ccaacatagt
     4741 aagccagtat acactccgct agcgctgatg tccggcggtg cttttgccgt tacgcaccac
     4801 cccgtcatga gattacgcgt tttagcggtg cgtacaatta agggattatg gtaaatccac
     4861 ttactgtctg ccctcgtagc catcgagata aaccgcagta ctccggccac gatgcgtccg
     4921 gcgtagagga tcgagatctt caatattggc cattagccat attattcatt ggttatatag
     4981 cataaatcaa tattggctat tggccattgc atacgttgta tctatatcat aatatgtaca
     5041 tttatattgg ctcatgtcca atatgaccgc catgttggca ttgattattg actagttatt
     5101 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
     5161 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
     5221 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
     5281 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtccgc
     5341 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
     5401 tacgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
     5461 tgcggttttg gcagtacacc aatgggcgtg gatagcggtt tgactca
//
