LOCUS       sequence-474729-        7030 bp    DNA     circular SYN 05-JUN-2025
DEFINITION  Express NanoLuc(R)-DGKB 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 7030)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 7030)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 5, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: NanoLuc-DGKb Fusion Vector
FEATURES             Location/Qualifiers
     source          1..7030
                     /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(4109..4127)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    4164..4285
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4175..4194)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4229..4248
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4530..4887
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4530..4721
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4553..4573)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4738..4873
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4902..4949
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4981..5775
                     /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(5035..5054)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5645..5664
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5839..5887
                     /note="synthetic polyadenylation signal"
     rep_origin      6044..6589
                     /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     6490..6509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6706..6723
                     /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 gacaaaccag gaaaaatggg
     1621 cccacctcag cccttcggaa ttttcccaac ttcagaaata tgctgagtat tctacaaaga
     1681 aattaaagga tgttcttgaa gaattccatg gtaatggtgt gcttgcaaag tataatcctg
     1741 aagggaaaca agacattctt aaccaaacaa tagattttga aggtttcaaa ctattcatga
     1801 agacattcct ggaagccgag cttcctgatg atttcactgc acaccttttc atgtcattta
     1861 gcaacaagtt tcctcattct agtccaatgg taaaaagtaa gcctgctctc ctatcaggcg
     1921 gtctgagaat gaataaaggt gccatcaccc ctccccggac tacttctcct gcaaatacgt
     1981 gttccccaga agtaatccat ctgaaggaca ttgtctgtta cctgtctctg cttgaaagag
     2041 gaagacctga ggataagctt gagtttatgt ttcgccttta tgacacggat gggaatggct
     2101 tcctggacag ctcggagcta gaaaatatca tcagtcagat gatgcatgtt gcagaatacc
     2161 ttgagtggga tgtcactgaa cttaatccaa tcctccatga aatgatggaa gaaattgact
     2221 atgatcatga tggaaccgtg tctcaaggag gaatgacaac gattccactt cttgtgctcc
     2281 tgggcttaga aaataacgtg aaggatgatg gacagcacgt gtggcgactg aagcacttta
     2341 acaaacctgc ctattgcaac ctttgcctga acatgctgat tggcgtgggg aagcagggcc
     2401 tctgctgttc cttctgcaag tacacagtcc atgagcgctg tgtggctcga gcacctccct
     2461 cttgcatcaa gacctatgtg aagtccaaaa ggaacactga tgtcatgcac cattactggg
     2521 ttgaaggtaa ctgcccaacc aagtgtgata agtgccacaa aactgttaaa tgttaccagg
     2581 gcctgacagg actgcattgt gtttggtgtc agatcacact gcataataaa tgtgcttctc
     2641 atctaaaacc tgaatgtgac tgtggacctt tgaaggacca tattttacca cccacaacaa
     2701 tctgtccagt ggtactgcag actctgccca cttcaggagt ttcagttcct gaggaaagac
     2761 aatcaacagt gaaaaaggaa aagagtggtt cccagcagcc aaacaaagtg attgacaaga
     2821 ataaaatgca aagagccaac tctgttactg tagatggaca aggcctgcag gtcactcctg
     2881 tgcctggtac tcacccactt ttagtttttg tgaaccccaa aagtggtgga aaacaaggag
     2941 aacgaattta cagaaaattc cagtatctat taaatcctcg tcaggtttac agtctttctg
     3001 gaaatggacc aatgccaggg ttaaactttt tccgtgatgt tcctgacttc agagtgttag
     3061 cctgtggtgg agatggaacc gtgggctggg ttttggattg catagaaaag gccaatgtag
     3121 gcaagcatcc tccagttgcg attctgcctc ttgggactgg caatgatcta gcaagatgcc
     3181 tgcgatgggg aggaggttac gaaggtgaga atctgatgaa aattctaaaa gacattgaaa
     3241 acagcacaga aatcatgttg gacaggtgga agtttgaagt catacctaat gacaaagatg
     3301 agaaaggaga cccagtgcct tacagtatca tcaataatta cttttccatt ggcgtggatg
     3361 cctccattgc acacagattc cacatcatga gagaaaaaca cccagagaaa ttcaacagta
     3421 gaatgaagaa caaattttgg tattttgagt ttggcacatc tgaaactttc tcagccacct
     3481 gcaagaagct acatgaatct gtagaaatag aatgtgatgg agtacagata gatttaataa
     3541 acatctctct ggaaggaatt gctattttga atataccaag catgcatgga ggatccaatc
     3601 tttggggaga gtctaagaaa agacgaagcc atcgacgaat agagaaaaaa gggtctgaca
     3661 aaaggaccac cgtcacagat gccaaagagt tgaagtttgc aagtcaagat ctcagtgacc
     3721 agctgctgga ggtggtcggc ttggaaggag ccatggagat ggggcaaata tacacaggcc
     3781 tgaaaagtgc tggccggcgg ctggctcagt gctcctgcgt ggtcatcagg acgagcaagt
     3841 ctctgccaat gcaaattgat ggggagccat ggatgcagac cccatgcaca ataaaaatta
     3901 cacacaagaa ccaagcccca atgctgatgg gcccgcctcc aaaaaccggt ttattctgct
     3961 ccctcgtcaa aaggacaaga aaccgaagca aggaagttta aacgaattcg ggctcggtac
     4021 ccggggatcc tctagagtcg acctgcaggc atgcaagctg atccggctgc taacaaagcc
     4081 cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg
     4141 gcggccgctt cgagcagaca tgataagata cattgatgag tttggacaaa ccacaactag
     4201 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     4261 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     4321 tcagggggag atgtgggagg tttttttaag caagtaaaac ctctacaaat gtggtaaaat
     4381 cgaattttaa caaaatatta acgcttacaa tttcctgatg cggtattttc tccttacgca
     4441 tctgtgcggt atttcacacc gcatacgcgg atctgcgcag caccatggcc tgaaataacc
     4501 tctgaaagag gaacttggtt aggtaccttc tgaggcggaa agaaccagct gtggaatgtg
     4561 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     4621 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     4681 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     4741 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     4801 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     4861 ttttttggag gcctaggctt ttgcaaaaag cttaattaac tgttgacaat taatcatcgg
     4921 catagtatat cggcatagta taatacgaca aggtgaggaa ctaaacccag gaggcagatc
     4981 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     5041 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     5101 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     5161 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     5221 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     5281 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     5341 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     5401 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     5461 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     5521 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     5581 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     5641 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     5701 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     5761 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     5821 tgccatcacg atggccgcaa taaaatatct ttattttcat tacatctgtg tgttggtttt
     5881 ttgtgtgaat cgatagcgat aaggatccgc tcatgcgatg ataagctgtc aaacatgaga
     5941 attacaactt atatcgtatg gggctgactt caggtgctac atttgaagag ataaattgca
     6001 ctgaaatcta gaaatatttt atctgattaa taagatgatc ttcttgagat cgttttggtc
     6061 tgcgcgtaat ctcttgctct gaaaacgaaa aaaccgcctt gcagggcggt ttttcgaagg
     6121 ttctctgagc taccaactct ttgaaccgag gtaactggct tggaggagcg cagtcaccaa
     6181 aacttgtcct ttcagtttag ccttaaccgg cgcatgactt caagactaac tcctctaaat
     6241 caattaccag tggctgctgc cagtggtgct tttgcatgtc tttccgggtt ggactcaaga
     6301 cgatagttac cggataaggc gcagcggtcg gactgaacgg ggggttcgtg catacagtcc
     6361 agcttggagc gaactgccta cccggaactg agtgtcaggc gtggaatgag acaaacgcgg
     6421 ccataacagc ggaatgacac cggtaaaccg aaaggcagga acaggagagc gcacgaggga
     6481 gccgccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc accactgatt
     6541 tgagcgtcag atttcgtgat gcttgtcagg ggggcggagc ctatggaaaa acggctttgc
     6601 cgcggccctc tcacttccct gttaagtatc ttcctggcat cttccaggaa atctccgccc
     6661 cgttcgtaag ccatttccgc tcgccgcagt cgaacgaccg agcgtagcga gtcagtgagc
     6721 gaggaagcgg aatatatcct gtatcacata ttctgctgac gcaccggtgc agcctttttt
     6781 ctcctgccac atgaagcact tcactgacac cctcatcagt gccaacatag taagccagta
     6841 tacactccgc tagcgctgat gtccggcggt gcttttgccg ttacgcacca ccccgtcatg
     6901 agattacgcg ttttagcggt gcgtacaatt aagggattat ggtaaatcca cttactgtct
     6961 gccctcgtag ccatcgagat aaaccgcagt actccggcca cgatgcgtcc ggcgtagagg
     7021 atcgagatct
//
