LOCUS       sequence-472936-        7006 bp    DNA     circular SYN 15-MAY-2025
DEFINITION  Express NanoLuc(R)-DGKG 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 7006)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 7006)
  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-DGKg Fusion Vector
FEATURES             Location/Qualifiers
     source          1..7006
                     /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(4085..4103)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    4140..4261
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4151..4170)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4205..4224
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4506..4863
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4506..4697
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4529..4549)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4714..4849
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4878..4925
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4957..5751
                     /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(5011..5030)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5621..5640
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5815..5863
                     /note="synthetic polyadenylation signal"
     rep_origin      6020..6565
                     /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     6466..6485
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6682..6699
                     /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 gggtgaagaa cggtgggtct
     1621 ccctcactcc agaagaattt gaccaactcc agaaatattc agaatattcc tccaagaaga
     1681 taaaagatgc cttgactgaa tttaatgagg gtgggagcct caaacaatat gacccacatg
     1741 agccgattag ctatgatgtc ttcaagctgt tcatgagggc gtacctggag gtggaccttc
     1801 cccagccact gagcactcac ctcttcctgg ccttcagcca gaagcccaga cacgagacct
     1861 ctgaccaccc gacggaggga gccagcaaca gtgaggccaa cagcgcagat actaatatac
     1921 agaatgcaga taatgccacc aaagcagacg aggcctgtgc ccctgatact gaatcaaata
     1981 tggctgagaa gcaagcacca gctgaagacc aagtggctgc gacccccctg gaaccccccg
     2041 tccctcggtc ttcaagctcg gaatccccag tggtatacct gaaggatgtt gtgtgctacc
     2101 tgtccctgct ggagacgggg aggcctcagg ataagctgga gttcatgttt cgcctctatg
     2161 attcagatga gaacggtctc ctggaccaag cggagatgga ttgcattgtc aaccaaatgc
     2221 tgcatattgc ccagtacctg gagtgggatc ccacagagct gaggcctata ttgaaggaga
     2281 tgctgcaagg gatggactac gaccgggacg gctttgtgtc tctacaggaa tgggtccatg
     2341 gagggatgac caccatccca ttgctggtcc tcctggggat ggatgactct ggctccaagg
     2401 gggatgggcg gcacgcctgg accatgaagc acttcaagaa accaacctac tgcaacttct
     2461 gccatatcat gctcatgggc gtccgcaagc aaggcctgtg ctgcacttac tgtaaataca
     2521 ctgtccacga acgctgtgtg tccagaaaca ttcctggttg tgtcaaaacg tactcaaaag
     2581 ccaaaaggag tggtgaggtg atgcagcacg catgggtgga agggaactcc tccgtcaagt
     2641 gtgaccggtg ccacaaaagt atcaagtgct accagagtgt caccgcgcgg cactgcgtgt
     2701 ggtgccggat gacgtttcac cgcaaatgtg aattatcaac gttgtgtgac ggtggggaac
     2761 tcagagacca catcttactg cccacctcca tatgccccat cacccgggac aggccaggtg
     2821 agaagtctga tggctgcgtg tccgccaagg gcgaacttgt catgcagtat aagatcatcc
     2881 ccaccccggg tacccacccc ctgctggtct tggtgaaccc caagagtgga gggagacaag
     2941 gagaaagaat tcttcggaaa ttccactatc tgctcaaccc caaacaagtt ttcaacctgg
     3001 acaatggggg gcctactcca gggttgaact ttttccgtga tactccagac ttccgtgttt
     3061 tggcctgtgg tggagatggg acagttggct ggattttgga ttgcattgat aaggccaact
     3121 ttgcaaagca tccaccagtg gctgtcctgc ctcttggaac aggaaatgac cttgcccgtt
     3181 gtctccgctg gggaggaggt tatgaagggg gcagcttgac aaaaatcctg aaagacattg
     3241 agcagagccc cttggtgatg ctggaccgct ggcatctgga agtcatcccc agagaggaag
     3301 tggaaaacgg ggaccaggtc ccatacagca tcatgaacaa ctatttctcc attggtgtgg
     3361 acgcttccat tgcacacaga ttccatgtga tgagagagaa acatcctgaa aaattcaaca
     3421 gcaggatgaa gaacaagctg tggtactttg aatttggcac ctcggagact tttgcagcga
     3481 cctgcaagaa actccacgac cacattgagt tggagtgtga tggggttggg gtggacctga
     3541 gcaacatctt cctggaaggc attgccattc tcaacattcc cagcatgtac ggaggcacca
     3601 atctctgggg agaaaacaag aagaaccggg ctgtgatccg ggaaagcagg aagggtgtca
     3661 ctgaccccaa agaactgaaa ttctgcgttc aagacctcag tgaccagctc cttgaagtgg
     3721 tggggctaga aggagccatg gagatggggc agatctacac cggcctgaag agtgcaggca
     3781 ggaggctggc ccagtgcgcc tctgtcacca tcaggacaaa caagctgctg ccaatgcaag
     3841 tggatggaga accctggatg cagccatgtt gcacgattaa aattactcac aagaaccaag
     3901 cgcccatgat gatggggcct ccccagaaga gcagcttctt ctcgttgaga aggaagagcc
     3961 gttcaaaaga cgtttaaacg aattcgggct cggtacccgg ggatcctcta gagtcgacct
     4021 gcaggcatgc aagctgatcc ggctgctaac aaagcccgaa aggaagctga gttggctgct
     4081 gccaccgctg agcaataact agcataaccc cttggggcgg ccgcttcgag cagacatgat
     4141 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     4201 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     4261 taacaacaac aattgcattc attttatgtt tcaggttcag ggggagatgt gggaggtttt
     4321 tttaagcaag taaaacctct acaaatgtgg taaaatcgaa ttttaacaaa atattaacgc
     4381 ttacaatttc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     4441 acgcggatct gcgcagcacc atggcctgaa ataacctctg aaagaggaac ttggttaggt
     4501 accttctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     4561 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     4621 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4681 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     4741 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     4801 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     4861 aaaaagctta attaactgtt gacaattaat catcggcata gtatatcggc atagtataat
     4921 acgacaaggt gaggaactaa acccaggagg cagatcatga ttgaacaaga tggattgcac
     4981 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     5041 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     5101 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     5161 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     5221 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     5281 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     5341 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     5401 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     5461 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     5521 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     5581 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     5641 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     5701 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     5761 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgatgg ccgcaataaa
     5821 atatctttat tttcattaca tctgtgtgtt ggttttttgt gtgaatcgat agcgataagg
     5881 atccgctcat gcgatgataa gctgtcaaac atgagaatta caacttatat cgtatggggc
     5941 tgacttcagg tgctacattt gaagagataa attgcactga aatctagaaa tattttatct
     6001 gattaataag atgatcttct tgagatcgtt ttggtctgcg cgtaatctct tgctctgaaa
     6061 acgaaaaaac cgccttgcag ggcggttttt cgaaggttct ctgagctacc aactctttga
     6121 accgaggtaa ctggcttgga ggagcgcagt caccaaaact tgtcctttca gtttagcctt
     6181 aaccggcgca tgacttcaag actaactcct ctaaatcaat taccagtggc tgctgccagt
     6241 ggtgcttttg catgtctttc cgggttggac tcaagacgat agttaccgga taaggcgcag
     6301 cggtcggact gaacgggggg ttcgtgcata cagtccagct tggagcgaac tgcctacccg
     6361 gaactgagtg tcaggcgtgg aatgagacaa acgcggccat aacagcggaa tgacaccggt
     6421 aaaccgaaag gcaggaacag gagagcgcac gagggagccg ccagggggaa acgcctggta
     6481 tctttatagt cctgtcgggt ttcgccacca ctgatttgag cgtcagattt cgtgatgctt
     6541 gtcagggggg cggagcctat ggaaaaacgg ctttgccgcg gccctctcac ttccctgtta
     6601 agtatcttcc tggcatcttc caggaaatct ccgccccgtt cgtaagccat ttccgctcgc
     6661 cgcagtcgaa cgaccgagcg tagcgagtca gtgagcgagg aagcggaata tatcctgtat
     6721 cacatattct gctgacgcac cggtgcagcc ttttttctcc tgccacatga agcacttcac
     6781 tgacaccctc atcagtgcca acatagtaag ccagtataca ctccgctagc gctgatgtcc
     6841 ggcggtgctt ttgccgttac gcaccacccc gtcatgagat tacgcgtttt agcggtgcgt
     6901 acaattaagg gattatggta aatccactta ctgtctgccc tcgtagccat cgagataaac
     6961 cgcagtactc cggccacgat gcgtccggcg tagaggatcg agatct
//
