LOCUS       sequence-473334-        6061 bp    DNA     circular SYN 14-JUL-2025
DEFINITION  Express NanoLuc(R)-EGR2 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 6061)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 6061)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: NanoLuc-EGR2 Fusion Vector
FEATURES             Location/Qualifiers
     source          1..6061
                     /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(3140..3158)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3195..3316
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3206..3225)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3260..3279
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3561..3918
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3561..3752
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3584..3604)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3769..3904
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3933..3980
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4012..4806
                     /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(4066..4085)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4676..4695
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4870..4918
                     /note="synthetic polyadenylation signal"
     rep_origin      5075..5620
                     /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     5521..5540
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5737..5754
                     /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 gatgacagcc aaggctgtgg
     1621 acaaaattcc tgtcactctc agtggttttg ttcaccagct tagtgataac atttatccag
     1681 tcgaggatct tgctgccact tctgtcacca tctttccaaa tgcggaactt ggggggccat
     1741 tcgatcagat gaacggtgta gccggggatg gaatgatcaa tatagacatg actggcgaga
     1801 agcgatctct tgacctgcca tacccgagta gcttcgcgcc tgtctctgcc ccgcgcaatc
     1861 agacatttac ttatatggga aaattttcaa tcgaccctca gtaccctgga gcctcatgct
     1921 accctgaggg gattataaat atagtgtccg ccggcatatt gcaaggtgta acgagcccgg
     1981 catcaactac cgcgagtagc tctgtgacga gcgcgagccc aaaccccctt gctaccgggc
     2041 cattgggtgt ctgtaccatg agtcaaactc aacctgacct tgaccatttg tacagtccac
     2101 cgcctccacc tcccccttac agcggatgtg cgggagattt gtaccaggat ccttctgcat
     2161 ttctgagtgc agctactact agcacatctt cttctttggc ttatcctccg ccaccttcct
     2221 acccctcacc gaaacccgca actgatcctg gtctctttcc catgatacct gactatcctg
     2281 ggttttttcc gagccagtgt cagcgcgatc tgcacggtac cgcaggacct gaccgaaagc
     2341 ccttcccttg tccactcgat acattgcgag taccaccacc gttgacccct cttagtacaa
     2401 tcaggaattt taccttggga ggtccatcag ctggagtgac gggacctggg gcgtccggtg
     2461 gttcagaagg tccgcggttg ccggggtcta gttcagccgc agcggcagct gctgcggcgg
     2521 cggcatataa tcctcatcat ttgccactgc gcccaattct gcggccacgc aaatacccaa
     2581 ataggcctag caaaacgccc gttcatgaaa gaccctaccc ttgtcccgcg gaagggtgcg
     2641 atcgaaggtt cagtcggtcc gacgagctca cccgccacat cagaatacac actggccata
     2701 agccgtttca gtgccggatt tgcatgcgga atttttcccg atcagatcac cttacaacac
     2761 atatccggac tcatacaggt gagaagccct ttgcttgtga ttattgtggc agaaagttcg
     2821 ccagaagtga tgaaaggaag cggcacacca aaattcacct tagacagaaa gagcgcaaat
     2881 catctgcgcc cagtgctagt gtccccgcac cgagtacggc ctcctgcagc ggtggtgtgc
     2941 agcctggggg cactctctgc agttcaaact ctagttctct cgggggggga ccgcttgccc
     3001 cttgttcctc aaggactcgg acaccagttt aaacgaattc gggctcggta cccggggatc
     3061 ctctagagtc gacctgcagg catgcaagct gatccggctg ctaacaaagc ccgaaaggaa
     3121 gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcggccgct
     3181 tcgagcagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     3241 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     3301 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga
     3361 gatgtgggag gtttttttaa gcaagtaaaa cctctacaaa tgtggtaaaa tcgaatttta
     3421 acaaaatatt aacgcttaca atttcctgat gcggtatttt ctccttacgc atctgtgcgg
     3481 tatttcacac cgcatacgcg gatctgcgca gcaccatggc ctgaaataac ctctgaaaga
     3541 ggaacttggt taggtacctt ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta
     3601 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3661 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3721 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     3781 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     3841 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     3901 ggcctaggct tttgcaaaaa gcttaattaa ctgttgacaa ttaatcatcg gcatagtata
     3961 tcggcatagt ataatacgac aaggtgagga actaaaccca ggaggcagat catgattgaa
     4021 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     4081 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     4141 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     4201 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     4261 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     4321 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     4381 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     4441 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     4501 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     4561 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     4621 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     4681 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     4741 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     4801 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     4861 gatggccgca ataaaatatc tttattttca ttacatctgt gtgttggttt tttgtgtgaa
     4921 tcgatagcga taaggatccg ctcatgcgat gataagctgt caaacatgag aattacaact
     4981 tatatcgtat ggggctgact tcaggtgcta catttgaaga gataaattgc actgaaatct
     5041 agaaatattt tatctgatta ataagatgat cttcttgaga tcgttttggt ctgcgcgtaa
     5101 tctcttgctc tgaaaacgaa aaaaccgcct tgcagggcgg tttttcgaag gttctctgag
     5161 ctaccaactc tttgaaccga ggtaactggc ttggaggagc gcagtcacca aaacttgtcc
     5221 tttcagttta gccttaaccg gcgcatgact tcaagactaa ctcctctaaa tcaattacca
     5281 gtggctgctg ccagtggtgc ttttgcatgt ctttccgggt tggactcaag acgatagtta
     5341 ccggataagg cgcagcggtc ggactgaacg gggggttcgt gcatacagtc cagcttggag
     5401 cgaactgcct acccggaact gagtgtcagg cgtggaatga gacaaacgcg gccataacag
     5461 cggaatgaca ccggtaaacc gaaaggcagg aacaggagag cgcacgaggg agccgccagg
     5521 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc caccactgat ttgagcgtca
     5581 gatttcgtga tgcttgtcag gggggcggag cctatggaaa aacggctttg ccgcggccct
     5641 ctcacttccc tgttaagtat cttcctggca tcttccagga aatctccgcc ccgttcgtaa
     5701 gccatttccg ctcgccgcag tcgaacgacc gagcgtagcg agtcagtgag cgaggaagcg
     5761 gaatatatcc tgtatcacat attctgctga cgcaccggtg cagccttttt tctcctgcca
     5821 catgaagcac ttcactgaca ccctcatcag tgccaacata gtaagccagt atacactccg
     5881 ctagcgctga tgtccggcgg tgcttttgcc gttacgcacc accccgtcat gagattacgc
     5941 gttttagcgg tgcgtacaat taagggatta tggtaaatcc acttactgtc tgccctcgta
     6001 gccatcgaga taaaccgcag tactccggcc acgatgcgtc cggcgtagag gatcgagatc
     6061 t
//
