LOCUS       sequence-470773-        8303 bp    DNA     circular SYN 15-MAY-2025
DEFINITION  Express NanoLuc(R)-DGKH 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 8303)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 8303)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: NanoLuc-DGKh Fusion Vector
FEATURES             Location/Qualifiers
     source          1..8303
                     /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(5372..5390)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    5427..5548
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5438..5457)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5492..5511
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        5793..6150
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5793..5984
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5816..5836)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6001..6136
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        6165..6212
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             6244..7038
                     /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(6298..6317)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6908..6927
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7112..7160
                     /note="synthetic polyadenylation signal"
     rep_origin      7317..7862
                     /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     7763..7782
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7979..7996
                     /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 ggcaggtgct ggaggacaac
     1621 atcatcctcc aggagctgct ggaggagcag cagctggagc tggtgcagca gttactagtg
     1681 cagctgcatc tgctggacca ggagaggatt cgtctgacag cgaagcggag caagagggac
     1741 cccagaaact gatccgcaaa gtgtctacct cggggcagat ccggaccaag accagtatta
     1801 aagagggaca gctattgaag caaaccagtt ctttccaaag gtggaaaaag cgatacttca
     1861 aacttcgagg ccgcaccctt tactatgcaa aggactcaaa gtctctgata tttgatgaag
     1921 ttgacctctc agatgctagt gtagctgaag caagcacgaa aaatgctaac aacagcttca
     1981 cgatcatcac tccattcaga aggctaatgc tgtgtgctga gaacagaaag gagatggagg
     2041 attggatcag ctcactgaag tctgtacaga ccagagaacc ctacgaggtg gcccagttta
     2101 atgtggaaca tttctcaggg atgcacaact ggtacgcctg ctcccacgcc cgacccacct
     2161 tctgtaacgt gtgcagagag agtctttctg gagtcacctc ccatggcctg tcctgcgaag
     2221 tgtgtaaatt caaggctcac aaaagatgtg cagtgagagc aacaaataac tgtaaatgga
     2281 ctaccctggc ctccatcggg aaggacatta tagaagatga agatggcgtc gcgatgcctc
     2341 accagtggct tgagggcaac ctgcctgtaa gtgccaagtg tgctgtctgc gacaaaacat
     2401 gtggcagtgt tctccgtcta caggattgga aatgcctttg gtgtaagaca atggtacaca
     2461 ctgcctgcaa agatttatac catccaatat gtccacttgg tcaatgtaaa gtatctatca
     2521 tacctccaat tgcactaaac agcaccgatt ccgatggttt ctgtagagca acattttcgt
     2581 tctgtgttag tcctctattg gtttttgtca attctaagag tggagataat cagggagtaa
     2641 agttcctccg tcgctttaaa cagttgctaa atccggctca ggtgtttgat ttaatgaatg
     2701 gaggtcctca tttaggttta agattatttc agaagtttga caatttccgg attcttgttt
     2761 gtggaggcga tggaagtgta ggttgggttt tgtcagaaat cgataagctc aacttgaata
     2821 aacagtgtca gctgggagtg ttgcctttgg gtacaggaaa tgaccttgcc cgagttcttg
     2881 gctggggagg ttcatatgac gatgacaccc aacttcctca gatcctagag aaactggaac
     2941 gagccagtac caaaatgttg gacaggtgga gtataatgac atatgaactc aaattgccac
     3001 caaaagcttc cctacttcca ggacctccag aagcatctga agaattttat atgacgattt
     3061 atgaagactc agttgcaacg catcttacaa aaatcctcaa ttctgatgaa catgcagtgg
     3121 tcatatcttc tgccaagacg ctatgtgaaa ctgtaaagga cttcgttgcc aaagtagaaa
     3181 agacgtatga caaaaccttg gaaaatgccg ttgtagctga tgccgtggcc agtaaatgtt
     3241 cagtcctaaa cgagaagctc gaacaactgc tgcaggcttt gcacacagat tcccaggctg
     3301 cgcctgttct ccctggcctc agccctctca ttgtggaaga agatgctgtg gaatcgtcca
     3361 gtgaagagtc cctgggtgaa agcaaagagc agcttgggga tgacgttaca aaaccttcct
     3421 cccagaaagc cgtcaaacca agggaaatca tgttgcgggc aaatagttta aagaaagcag
     3481 tgaggcaagt cattgaggaa gccggaaaag ttatggatga cccgacagtt cacccctgtg
     3541 aaccagctaa tcagtcctct gattatgaca gcacagaaac agatgaatct aaggaggaag
     3601 ctaaagatga tggtgccaaa gaatcaataa ctgttaaaac tgcacctcgg tctccagatg
     3661 cccgggcaag ttatggccat tcccaaactg attctgtccc tggtccagct gtggcagcca
     3721 gcaaagaaaa cctccctgtg ctcaatacca gaataatctg cccaggttta agagcaggac
     3781 tggctgcctc aattgctggg agttcgatta tcaacaaaat gttactggca aacattgatc
     3841 ctttcggtgc cacgccgttt attgacccgg atctagattc cgtagatgga tattcagaaa
     3901 aatgtgtcat gaacaattac tttgggattg gattagatgc aaaaatttca ttagaattta
     3961 ataataaaag agaggagcac cctgaaaaat gcaggagccg aactaaaaac ttgatgtggt
     4021 atggagtcct tggaacccgg gagttattac agagatcgta caagaattta gaacaaaggg
     4081 ttcaacttga gtgtgatggg cagtatattc ctcttcccag cttgcaaggc atagccgtgt
     4141 tgaacattcc cagctatgct ggaggcacta acttttgggg tggaactaaa gaggatgata
     4201 tatttgctgc accatccttt gatgacaaga tcctggaagt tgtagcaata tttgatagca
     4261 tgcaaatggc agtttcaagg gtcattaaac tgcagcatca tcgaatagcc cagtgccgta
     4321 cagtgaaaat cactatattt ggtgacgaag gagtcccagt gcaagtggat ggtgaagcgt
     4381 gggttcagcc tccagggatt atcaaaattg tgcacaaaaa cagagcacaa atgctaacaa
     4441 gggacagagc ctttgagagc actctgaaat cttgggaaga taagcagaag tgtgattctg
     4501 gtaaaccagt tctccgaacc catttgtaca tccatcacgc cattgacttg gcaacagaag
     4561 aggtgtcgca gatgcagcta tgctcccagg ctgcagagga gctcattact aggatatgtg
     4621 acgcagccac aattcactgt cttttggagc aagaactggc ccatgctgtg aatgcctgct
     4681 cccatgccct gaataaggcc aacccaaggt gcccggagag tcttacaaga gacactgcca
     4741 ctgaaatagc catcaatgtg aaggcgctgt ataatgaaac agaatctttg ctagttggca
     4801 gggttccttt gcagctggaa tcgccacatg aagagcgagt atccaatgcc ttacactctg
     4861 tggaggtgga attacagaaa ctgacagaga ttccttggct ttattatatc ttacacccaa
     4921 atgaggatga ggaacctcct atggattgca ccaaaaggaa caacagaagc accgtatttc
     4981 gaatagtgcc aaagtttaaa aaggaaaagg ttcagaagca gaagacaagt tcacagcctg
     5041 ttcagaaatg gggcacagag gaagttgctg cttggctgga tctgctcaat ttgggagagt
     5101 acaaagatat cttcatccgt catgacatca gaggggctga acttttgcat ctggaaaggc
     5161 gagatcttaa ggatctgggg ataccgaaag tgggtcatgt gaagcgaatt ctccagggaa
     5221 ttaaagagct tggaaggagc actccacagt cggaggtggt ttaaacgaat tcgggctcgg
     5281 tacccgggga tcctctagag tcgacctgca ggcatgcaag ctgatccggc tgctaacaaa
     5341 gcccgaaagg aagctgagtt ggctgctgcc accgctgagc aataactagc ataacccctt
     5401 ggggcggccg cttcgagcag acatgataag atacattgat gagtttggac aaaccacaac
     5461 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
     5521 aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca
     5581 ggttcagggg gagatgtggg aggttttttt aagcaagtaa aacctctaca aatgtggtaa
     5641 aatcgaattt taacaaaata ttaacgctta caatttcctg atgcggtatt ttctccttac
     5701 gcatctgtgc ggtatttcac accgcatacg cggatctgcg cagcaccatg gcctgaaata
     5761 acctctgaaa gaggaacttg gttaggtacc ttctgaggcg gaaagaacca gctgtggaat
     5821 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     5881 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     5941 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     6001 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     6061 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     6121 ggcttttttg gaggcctagg cttttgcaaa aagcttaatt aactgttgac aattaatcat
     6181 cggcatagta tatcggcata gtataatacg acaaggtgag gaactaaacc caggaggcag
     6241 atcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     6301 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     6361 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     6421 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     6481 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     6541 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     6601 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     6661 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     6721 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     6781 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     6841 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     6901 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     6961 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     7021 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     7081 acctgccatc acgatggccg dgkgtnrtnm caataaaata tctttatttt cattacatct
     7141 gtgtgttggt tttttgtgtg aatcgatagc gataaggatc cgctcatgcg atgataagct
     7201 gtcaaacatg agaattacaa cttatatcgt atggggctga cttcaggtgc tacatttgaa
     7261 gagataaatt gcactgaaat ctagaaatat tttatctgat taataagatg atcttcttga
     7321 gatcgttttg gtctgcgcgt aatctcttgc tctgaaaacg aaaaaaccgc cttgcagggc
     7381 ggtttttcga aggttctctg agctaccaac tctttgaacc gaggtaactg gcttggagga
     7441 gcgcagtcac caaaacttgt cctttcagtt tagccttaac cggcgcatga cttcaagact
     7501 aactcctcta aatcaattac cagtggctgc tgccagtggt gcttttgcat gtctttccgg
     7561 gttggactca agacgatagt taccggataa ggcgcagcgg tcggactgaa cggggggttc
     7621 gtgcatacag tccagcttgg agcgaactgc ctacccggaa ctgagtgtca ggcgtggaat
     7681 gagacaaacg cggccataac agcggaatga caccggtaaa ccgaaaggca ggaacaggag
     7741 agcgcacgag ggagccgcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     7801 gccaccactg atttgagcgt cagatttcgt gatgcttgtc aggggggcgg agcctatgga
     7861 aaaacggctt tgccgcggcc ctctcacttc cctgttaagt atcttcctgg catcttccag
     7921 gaaatctccg ccccgttcgt aagccatttc cgctcgccgc agtcgaacga ccgagcgtag
     7981 cgagtcagtg agcgaggaag cggaatatat cctgtatcac atattctgct gacgcaccgg
     8041 tgcagccttt tttctcctgc cacatgaagc acttcactga caccctcatc agtgccaaca
     8101 tagtaagcca gtatacactc cgctagcgct gatgtccggc ggtgcttttg ccgttacgca
     8161 ccaccccgtc atgagattac gcgttttagc ggtgcgtaca attaagggat tatggtaaat
     8221 ccacttactg tctgccctcg tagccatcga gataaaccgc agtactccgg ccacgatgcg
     8281 tccggcgtag aggatcgaga tct
//
