LOCUS       sequence-480847-        5773 bp    DNA     circular SYN 28-JUL-2025
DEFINITION  Express MAP2K1-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 5773)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5773)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 28, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: MAP2K1-NanoLuc Fusion Vector
FEATURES             Location/Qualifiers
     source          1..5773
                     /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             2265..2777
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     primer_bind     complement(2865..2883)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2920..3041
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2931..2950)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2985..3004
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3286..3643
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3286..3477
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3309..3329)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3494..3629
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3724..4518
                     /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(3778..3797)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4388..4407
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4582..4630
                     /note="synthetic polyadenylation signal"
     rep_origin      4787..5332
                     /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     5233..5252
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5449..5466
                     /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 cgccatgccc aagaagaagc
     1081 cgacgcccat ccagctgaac ccggcccccg acggctctgc agttaacggg accagctctg
     1141 cggagaccaa cttggaggcc ttgcagaaga agctggagga gctagagctt gatgagcagc
     1201 agcgaaagcg ccttgaggcc tttcttaccc agaagcagaa ggtgggagaa ctgaaggatg
     1261 acgactttga gaagatcagt gagctggggg ctggcaatgg cggtgtggtg ttcaaggtct
     1321 cccacaagcc ttctggcctg gtcatggcca gaaagctaat tcatctggag atcaaacccg
     1381 caatccggaa ccagatcata agggagctgc aggttctgca tgagtgcaac tctccgtaca
     1441 tcgtgggctt ctatggtgcg ttctacagcg atggcgagat cagtatctgc atggagcaca
     1501 tggatggagg ttctctggat caagtcctga agaaagctgg aagaattcct gaacaaattt
     1561 taggaaaagt tagcattgct gtaataaaag gcctgacata tctgagggag aagcacaaga
     1621 tcatgcacag agatgtcaag ccctccaaca tcctagtcaa ctcccgtggg gagatcaagc
     1681 tctgtgactt tggggtcagc gggcagctca tcgactccat ggccaactcc ttcgtgggca
     1741 caaggtccta catgtcgcca gaaagactcc aggggactca ttactctgtg cagtcagaca
     1801 tctggagcat gggactgtct ctggtagaga tggcggttgg gaggtatccc atccctcctc
     1861 cagatgccaa ggagctggag ctgatgtttg ggtgccaggt ggaaggagat gcggctgaga
     1921 ccccacccag gccaaggacc cccgggaggc cccttagctc atacggaatg gacagccgac
     1981 ctcccatggc aatttttgag ttgttggatt acatagtcaa cgagcctcct ccaaaactgc
     2041 ccagtggagt gttcagtctg gaatttcaag attttgtgaa taaatgctta ataaaaaacc
     2101 ccgcagagag agcagatttg aagcaactca tggttcatgc ttttatcaag agatctgatg
     2161 ctgaggaagt ggattttgca ggttggctct gctccaccat cggccttaac cagcccagca
     2221 caccaaccca tgctgctggc gtcgtttctc tcggctcgag cggcgtcttc acactcgaag
     2281 atttcgttgg ggactggcga cagacagccg gctacaacct ggaccaagtc cttgaacagg
     2341 gaggtgtgtc cagtttgttt cagaatctcg gggtgtccgt aactccgatc caaaggattg
     2401 tcctgagcgg tgaaaatggg ctgaagatcg acatccatgt catcatcccg tatgaaggtc
     2461 tgagcggcga ccaaatgggc cagatcgaaa aaatttttaa ggtggtgtac cctgtggatg
     2521 atcatcactt taaggtgatc ctgcactatg gcacactggt aatcgacggg gttacgccga
     2581 acatgatcga ctatttcgga cggccgtatg aaggcatcgc cgtgttcgac ggcaaaaaga
     2641 tcactgtaac agggaccctg tggaacggca acaaaattat cgacgagcgc ctgatcaacc
     2701 ccgacggctc cctgctgttc cgagtaacca tcaacggagt gaccggctgg cggctgtgcg
     2761 aacgcattct ggcgtaaggc cgcgactcta gagtcgacct gcaggcatgc aagctgatcc
     2821 ggctgctaac aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact
     2881 agcataaccc cttggggcgg ccgcttcgag cagacatgat aagatacatt gatgagtttg
     2941 gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta
     3001 ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc
     3061 attttatgtt tcaggttcag ggggagatgt gggaggtttt tttaagcaag taaaacctct
     3121 acaaatgtgg taaaatcgaa ttttaacaaa atattaacgc ttacaatttc ctgatgcggt
     3181 attttctcct tacgcatctg tgcggtattt cacaccgcat acgcggatct gcgcagcacc
     3241 atggcctgaa ataacctctg aaagaggaac ttggttaggt accttctgag gcggaaagaa
     3301 ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     3361 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     3421 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     3481 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     3541 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     3601 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctta attaactgtt
     3661 gacaattaat catcggcata gtataatacg acaaggtgag gaactaaacc caggaggcag
     3721 atcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     3781 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     3841 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     3901 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     3961 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     4021 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     4081 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     4141 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     4201 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     4261 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     4321 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     4381 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     4441 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     4501 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     4561 acctgccatc acgatggccg caataaaata tctttatttt cattacatct gtgtgttggt
     4621 tttttgtgtg aatcgatagc gataaggatc cgctcatgcg atgataagct gtcaaacatg
     4681 agaattacaa cttatatcgt atggggctga cttcaggtgc tacatttgaa gagataaatt
     4741 gcactgaaat ctagaaatat tttatctgat taataagatg atcttcttga gatcgttttg
     4801 gtctgcgcgt aatctcttgc tctgaaaacg aaaaaaccgc cttgcagggc ggtttttcga
     4861 aggttctctg agctaccaac tctttgaacc gaggtaactg gcttggagga gcgcagtcac
     4921 caaaacttgt cctttcagtt tagccttaac cggcgcatga cttcaagact aactcctcta
     4981 aatcaattac cagtggctgc tgccagtggt gcttttgcat gtctttccgg gttggactca
     5041 agacgatagt taccggataa ggcgcagcgg tcggactgaa cggggggttc gtgcatacag
     5101 tccagcttgg agcgaactgc ctacccggaa ctgagtgtca ggcgtggaat gagacaaacg
     5161 cggccataac agcggaatga caccggtaaa ccgaaaggca ggaacaggag agcgcacgag
     5221 ggagccgcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccaccactg
     5281 atttgagcgt cagatttcgt gatgcttgtc aggggggcgg agcctatgga aaaacggctt
     5341 tgccgcggcc ctctcacttc cctgttaagt atcttcctgg catcttccag gaaatctccg
     5401 ccccgttcgt aagccatttc cgctcgccgc agtcgaacga ccgagcgtag cgagtcagtg
     5461 agcgaggaag cggaatatat cctgtatcac atattctgct gacgcaccgg tgcagccttt
     5521 tttctcctgc cacatgaagc acttcactga caccctcatc agtgccaaca tagtaagcca
     5581 gtatacactc cgctagcgct gatgtccggc ggtgcttttg ccgttacgca ccaccccgtc
     5641 atgagattac gcgttttagc ggtgcgtaca attaagggat tatggtaaat ccacttactg
     5701 tctgccctcg tagccatcga gataaaccgc agtactccgg ccacgatgcg tccggcgtag
     5761 aggatcgaga tct
//
