LOCUS       sequence-478485-        6260 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Express EIF2AK2-NanoLuc 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 6260)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6260)
  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            "Unpublished"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: EIF2AK2-NanoLuc Fusion Vector
FEATURES             Location/Qualifiers
     source          1..6260
                     /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             2739..3251
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     primer_bind     complement(3339..3357)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3394..3515
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3405..3424)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3459..3478
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3760..4117
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3760..3951
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3783..3803)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3968..4103
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4132..4179
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4211..5005
                     /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(4265..4284)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4875..4894
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5069..5117
                     /note="synthetic polyadenylation signal"
     rep_origin      5274..5819
                     /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     5720..5739
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5936..5953
                     /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 cgccatggct ggtgatcttt
     1081 cagcaggttt cttcatggag gaacttaata cataccgtca gaagcaggga gtagtactta
     1141 aatatcaaga actgcctaat tcaggacctc cacatgatag gaggtttaca tttcaagtta
     1201 taatagatgg aagagaattt ccagaaggtg aaggtagatc aaagaaggaa gcaaaaaatg
     1261 ccgcagccaa attagctgtt gagatactta ataaggaaaa gaaggcagtt agtcctttat
     1321 tattgacaac aacgaattct tcagaaggat tatccatggg gaattacata ggccttatca
     1381 atagaattgc ccagaagaaa agactaactg taaattatga acagtgtgca tcgggggtgc
     1441 atgggccaga aggatttcat tataaatgca aaatgggaca gaaagaatat agtattggta
     1501 caggttctac taaacaggaa gcaaaacaat tggccgctaa acttgcatat cttcagatat
     1561 tatcagaaga aacctcagtg aaatctgact acctgtcctc tggttctttt gctactacgt
     1621 gtgagtccca aagcaactct ttagtgacca gcacactcgc ttctgaatca tcatctgaag
     1681 gtgacttctc agcagataca tcagagataa attctaacag tgacagtttg aacagttctt
     1741 cgttgcttat gaatggtctc agaaataatc aaaggaaggc aaaaagatct ttggcaccca
     1801 gatttgacct tcctgacatg aaagaaacaa agtatactgt ggacaagagg tttggcatgg
     1861 attttaaaga aatagaatta attggctcag gtggatttgg ccaagttttc aaagcaaaac
     1921 acagaattga cggaaagact tacgttatta aacgtgttaa atataataac gagaaggcgg
     1981 agcgtgaagt aaaagcattg gcaaaacttg atcatgtaaa tattgttcac tacaatggct
     2041 gttgggatgg atttgattat gatcctgaga ccagtgatga ttctcttgag agcagtgatt
     2101 atgatcctga gaacagcaaa aatagttcaa ggtcaaagac taagtgcctt ttcatccaaa
     2161 tggaattctg tgataaaggg accttggaac aatggattga aaaaagaaga ggcgagaaac
     2221 tagacaaagt tttggctttg gaactctttg aacaaataac aaaaggggtg gattatatac
     2281 attcaaaaaa attaattcat agagatctta agccaagtaa tatattctta gtagatacaa
     2341 aacaagtaaa gattggagac tttggacttg taacatctct gaaaaatgat ggaaagcgaa
     2401 caaggagtaa gggaactttg cgatacatga gcccagaaca gatttcttcg caagactatg
     2461 gaaaggaagt ggacctctac gctttggggc taattcttgc tgaacttctt catgtatgtg
     2521 acactgcttt tgaaacatca aagtttttca cagacctacg ggatggcatc atctcagata
     2581 tatttgataa aaaagaaaaa actcttctac agaaattact ctcaaagaaa cctgaggatc
     2641 gacctaacac atctgaaata ctaaggacct tgactgtgtg gaagaaaagc ccagagaaaa
     2701 atgaacgaca cacatgtgtt tctctcggct cgagcggcgt cttcacactc gaagatttcg
     2761 ttggggactg gcgacagaca gccggctaca acctggacca agtccttgaa cagggaggtg
     2821 tgtccagttt gtttcagaat ctcggggtgt ccgtaactcc gatccaaagg attgtcctga
     2881 gcggtgaaaa tgggctgaag atcgacatcc atgtcatcat cccgtatgaa ggtctgagcg
     2941 gcgaccaaat gggccagatc gaaaaaattt ttaaggtggt gtaccctgtg gatgatcatc
     3001 actttaaggt gatcctgcac tatggcacac tggtaatcga cggggttacg ccgaacatga
     3061 tcgactattt cggacggccg tatgaaggca tcgccgtgtt cgacggcaaa aagatcactg
     3121 taacagggac cctgtggaac ggcaacaaaa ttatcgacga gcgcctgatc aaccccgacg
     3181 gctccctgct gttccgagta accatcaacg gagtgaccgg ctggcggctg tgcgaacgca
     3241 ttctggcgta aggccgcgac tctagagtcg acctgcaggc atgcaagctg atccggctgc
     3301 taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactagcata
     3361 accccttggg gcggccgctt cgagcagaca tgataagata cattgatgag tttggacaaa
     3421 ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt
     3481 tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta
     3541 tgtttcaggt tcagggggag atgtgggagg tttttttaag caagtaaaac ctctacaaat
     3601 gtggtaaaat cgaattttaa caaaatatta acgcttacaa tttcctgatg cggtattttc
     3661 tccttacgca tctgtgcggt atttcacacc gcatacgcgg atctgcgcag caccatggcc
     3721 tgaaataacc tctgaaagag gaacttggtt aggtaccttc tgaggcggaa agaaccagct
     3781 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     3841 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     3901 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     3961 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     4021 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     4081 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag cttaattaac tgttgacaat
     4141 taatcatcgg catagtatat cggcatagta taatacgaca aggtgaggaa ctaaacccag
     4201 gaggcagatc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     4261 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     4321 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     4381 gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     4441 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     4501 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     4561 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     4621 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     4681 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg
     4741 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     4801 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     4861 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     4921 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     4981 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     5041 acgcccaacc tgccatcacg atggccgcaa taaaatatct ttattttcat tacatctgtg
     5101 tgttggtttt ttgtgtgaat cgatagcgat aaggatccgc tcatgcgatg ataagctgtc
     5161 aaacatgaga attacaactt atatcgtatg gggctgactt caggtgctac atttgaagag
     5221 ataaattgca ctgaaatcta gaaatatttt atctgattaa taagatgatc ttcttgagat
     5281 cgttttggtc tgcgcgtaat ctcttgctct gaaaacgaaa aaaccgcctt gcagggcggt
     5341 ttttcgaagg ttctctgagc taccaactct ttgaaccgag gtaactggct tggaggagcg
     5401 cagtcaccaa aacttgtcct ttcagtttag ccttaaccgg cgcatgactt caagactaac
     5461 tcctctaaat caattaccag tggctgctgc cagtggtgct tttgcatgtc tttccgggtt
     5521 ggactcaaga cgatagttac cggataaggc gcagcggtcg gactgaacgg ggggttcgtg
     5581 catacagtcc agcttggagc gaactgccta cccggaactg agtgtcaggc gtggaatgag
     5641 acaaacgcgg ccataacagc ggaatgacac cggtaaaccg aaaggcagga acaggagagc
     5701 gcacgaggga gccgccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     5761 accactgatt tgagcgtcag atttcgtgat gcttgtcagg ggggcggagc ctatggaaaa
     5821 acggctttgc cgcggccctc tcacttccct gttaagtatc ttcctggcat cttccaggaa
     5881 atctccgccc cgttcgtaag ccatttccgc tcgccgcagt cgaacgaccg agcgtagcga
     5941 gtcagtgagc gaggaagcgg aatatatcct gtatcacata ttctgctgac gcaccggtgc
     6001 agcctttttt ctcctgccac atgaagcact tcactgacac cctcatcagt gccaacatag
     6061 taagccagta tacactccgc tagcgctgat gtccggcggt gcttttgccg ttacgcacca
     6121 ccccgtcatg agattacgcg ttttagcggt gcgtacaatt aagggattat ggtaaatcca
     6181 cttactgtct gccctcgtag ccatcgagat aaaccgcagt actccggcca cgatgcgtcc
     6241 ggcgtagagg atcgagatct
//
