LOCUS       sequence-479795-        6286 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Express NanoLuc-EIF2AK2 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 6286)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6286)
  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            "Unpublished"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-EIF2AK2 Fusion Vector
FEATURES             Location/Qualifiers
     source          1..6286
                     /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(3365..3383)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3420..3541
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3431..3450)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3485..3504
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3786..4143
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3786..3977
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3809..3829)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3994..4129
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4158..4205
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4237..5031
                     /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(4291..4310)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4901..4920
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5095..5143
                     /note="synthetic polyadenylation signal"
     rep_origin      5300..5845
                     /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     5746..5765
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5962..5979
                     /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 ggctggtgat ctttcagcag
     1621 gtttcttcat ggaggaactt aatacatacc gtcagaagca gggagtagta cttaaatatc
     1681 aagaactgcc taattcagga cctccacatg ataggaggtt tacatttcaa gttataatag
     1741 atggaagaga atttccagaa ggtgaaggta gatcaaagaa ggaagcaaaa aatgccgcag
     1801 ccaaattagc tgttgagata cttaataagg aaaagaaggc agttagtcct ttattattga
     1861 caacaacgaa ttcttcagaa ggattatcca tggggaatta cataggcctt atcaatagaa
     1921 ttgcccagaa gaaaagacta actgtaaatt atgaacagtg tgcatcgggg gtgcatgggc
     1981 cagaaggatt tcattataaa tgcaaaatgg gacagaaaga atatagtatt ggtacaggtt
     2041 ctactaaaca ggaagcaaaa caattggccg ctaaacttgc atatcttcag atattatcag
     2101 aagaaacctc agtgaaatct gactacctgt cctctggttc ttttgctact acgtgtgagt
     2161 cccaaagcaa ctctttagtg accagcacac tcgcttctga atcatcatct gaaggtgact
     2221 tctcagcaga tacatcagag ataaattcta acagtgacag tttgaacagt tcttcgttgc
     2281 ttatgaatgg tctcagaaat aatcaaagga aggcaaaaag atctttggca cccagatttg
     2341 accttcctga catgaaagaa acaaagtata ctgtggacaa gaggtttggc atggatttta
     2401 aagaaataga attaattggc tcaggtggat ttggccaagt tttcaaagca aaacacagaa
     2461 ttgacggaaa gacttacgtt attaaacgtg ttaaatataa taacgagaag gcggagcgtg
     2521 aagtaaaagc attggcaaaa cttgatcatg taaatattgt tcactacaat ggctgttggg
     2581 atggatttga ttatgatcct gagaccagtg atgattctct tgagagcagt gattatgatc
     2641 ctgagaacag caaaaatagt tcaaggtcaa agactaagtg ccttttcatc caaatggaat
     2701 tctgtgataa agggaccttg gaacaatgga ttgaaaaaag aagaggcgag aaactagaca
     2761 aagttttggc tttggaactc tttgaacaaa taacaaaagg ggtggattat atacattcaa
     2821 aaaaattaat tcatagagat cttaagccaa gtaatatatt cttagtagat acaaaacaag
     2881 taaagattgg agactttgga cttgtaacat ctctgaaaaa tgatggaaag cgaacaagga
     2941 gtaagggaac tttgcgatac atgagcccag aacagatttc ttcgcaagac tatggaaagg
     3001 aagtggacct ctacgctttg gggctaattc ttgctgaact tcttcatgta tgtgacactg
     3061 cttttgaaac atcaaagttt ttcacagacc tacgggatgg catcatctca gatatatttg
     3121 ataaaaaaga aaaaactctt ctacagaaat tactctcaaa gaaacctgag gatcgaccta
     3181 acacatctga aatactaagg accttgactg tgtggaagaa aagcccagag aaaaatgaac
     3241 gacacacatg tgtttaaacg aattcgggct cggtacccgg ggatcctcta gagtcgacct
     3301 gcaggcatgc aagctgatcc ggctgctaac aaagcccgaa aggaagctga gttggctgct
     3361 gccaccgctg agcaataact agcataaccc cttggggcgg ccgcttcgag cagacatgat
     3421 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     3481 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     3541 taacaacaac aattgcattc attttatgtt tcaggttcag ggggagatgt gggaggtttt
     3601 tttaagcaag taaaacctct acaaatgtgg taaaatcgaa ttttaacaaa atattaacgc
     3661 ttacaatttc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     3721 acgcggatct gcgcagcacc atggcctgaa ataacctctg aaagaggaac ttggttaggt
     3781 accttctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     3841 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     3901 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     3961 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     4021 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     4081 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     4141 aaaaagctta attaactgtt gacaattaat catcggcata gtatatcggc atagtataat
     4201 acgacaaggt gaggaactaa acccaggagg cagatcatga ttgaacaaga tggattgcac
     4261 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     4321 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     4381 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     4441 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     4501 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     4561 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     4621 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     4681 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     4741 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     4801 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     4861 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     4921 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     4981 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     5041 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgatgg ccgcaataaa
     5101 atatctttat tttcattaca tctgtgtgtt ggttttttgt gtgaatcgat agcgataagg
     5161 atccgctcat gcgatgataa gctgtcaaac atgagaatta caacttatat cgtatggggc
     5221 tgacttcagg tgctacattt gaagagataa attgcactga aatctagaaa tattttatct
     5281 gattaataag atgatcttct tgagatcgtt ttggtctgcg cgtaatctct tgctctgaaa
     5341 acgaaaaaac cgccttgcag ggcggttttt cgaaggttct ctgagctacc aactctttga
     5401 accgaggtaa ctggcttgga ggagcgcagt caccaaaact tgtcctttca gtttagcctt
     5461 aaccggcgca tgacttcaag actaactcct ctaaatcaat taccagtggc tgctgccagt
     5521 ggtgcttttg catgtctttc cgggttggac tcaagacgat agttaccgga taaggcgcag
     5581 cggtcggact gaacgggggg ttcgtgcata cagtccagct tggagcgaac tgcctacccg
     5641 gaactgagtg tcaggcgtgg aatgagacaa acgcggccat aacagcggaa tgacaccggt
     5701 aaaccgaaag gcaggaacag gagagcgcac gagggagccg ccagggggaa acgcctggta
     5761 tctttatagt cctgtcgggt ttcgccacca ctgatttgag cgtcagattt cgtgatgctt
     5821 gtcagggggg cggagcctat ggaaaaacgg ctttgccgcg gccctctcac ttccctgtta
     5881 agtatcttcc tggcatcttc caggaaatct ccgccccgtt cgtaagccat ttccgctcgc
     5941 cgcagtcgaa cgaccgagcg tagcgagtca gtgagcgagg aagcggaata tatcctgtat
     6001 cacatattct gctgacgcac cggtgcagcc ttttttctcc tgccacatga agcacttcac
     6061 tgacaccctc atcagtgcca acatagtaag ccagtataca ctccgctagc gctgatgtcc
     6121 ggcggtgctt ttgccgttac gcaccacccc gtcatgagat tacgcgtttt agcggtgcgt
     6181 acaattaagg gattatggta aatccactta ctgtctgccc tcgtagccat cgagataaac
     6241 cgcagtactc cggccacgat gcgtccggcg tagaggatcg agatct
//
