LOCUS       sequence-470770-        7045 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 7045)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 7045)
  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..7045
                     /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(4124..4142)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    4179..4300
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4190..4209)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4244..4263
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4545..4902
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4545..4736
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4568..4588)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4753..4888
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4917..4964
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4996..5790
                     /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(5050..5069)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5660..5679
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5854..5902
                     /note="synthetic polyadenylation signal"
     rep_origin      6059..6604
                     /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     6505..6524
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6721..6738
                     /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 tctctggagg aatggattca aggaggaatg acaacgattc
     2281 cacttcttgt gctcctgggc ttagaaaata acgtgaagga tgatggacag cacgtgtggc
     2341 gactgaagca ctttaacaaa cctgcctatt gcaacctttg cctgaacatg ctgattggcg
     2401 tggggaagca gggcctctgc tgttccttct gcaagtacac agtccatgag cgctgtgtgg
     2461 ctcgagcacc tccctcttgc atcaagacct atgtgaagtc caaaaggaac actgatgtca
     2521 tgcaccatta ctgggttgaa ggtaactgcc caaccaagtg tgataagtgc cacaaaactg
     2581 ttaaatgtta ccagggcctg acaggactgc attgtgtttg gtgtcagatc acactgcata
     2641 ataaatgtgc ttctcatcta aaacctgaat gtgactgtgg acctttgaag gaccatattt
     2701 taccacccac aacaatctgt ccagtggtac tgcagactct gcccacttca ggagtttcag
     2761 ttcctgagga aagacaatca acagtgaaaa aggaaaagag tggttcccag cagccaaaca
     2821 aagtgattga caagaataaa atgcaaagag ccaactctgt tactgtagat ggacaaggcc
     2881 tgcaggtcac tcctgtgcct ggtactcacc cacttttagt ttttgtgaac cccaaaagtg
     2941 gtggaaaaca aggagaacga atttacagaa aattccagta tctattaaat cctcgtcagg
     3001 tttacagtct ttctggaaat ggaccaatgc cagggttaaa ctttttccgt gatgttcctg
     3061 acttcagagt gttagcctgt ggtggagatg gaaccgtggg ctgggttttg gattgcatag
     3121 aaaaggccaa tgtaggcaag catcctccag ttgcgattct gcctcttggg actggcaatg
     3181 atctagcaag atgcctgcga tggggaggag gttacgaagg tgagaatctg atgaaaattc
     3241 taaaagacat tgaaaacagc acagaaatca tgttggacag gtggaagttt gaagtcatac
     3301 ctaatgacaa agatgagaaa ggagacccag tgccttacag tatcatcaat aattactttt
     3361 ccattggcgt ggatgcctcc attgcacaca gattccacat catgagagaa aaacacccag
     3421 agaaattcaa cagtagaatg aagaacaaat tttggtattt tgagtttggc acatctgaaa
     3481 ctttctcagc cacctgcaag aagctacatg aatctgtaga aatagaatgt gatggagtac
     3541 agatagattt aataaacatc tctctggaag gaattgctat tttgaatata ccaagcatgc
     3601 atggaggatc caatctttgg ggagagtcta agaaaagacg aagccatcga cgaatagaga
     3661 aaaaagggtc tgacaaaagg accaccgtca cagatgccaa agagttgaag tttgcaagtc
     3721 aagatctcag tgaccagctg ctggaggtgg tcggcttgga aggagccatg gagatggggc
     3781 aaatatacac aggcctgaaa agtgctggcc ggcggctggc tcagtgctcc tgcgtggtca
     3841 tcaggacgag caagtctctg ccaatgcaaa ttgatgggga gccatggatg cagaccccat
     3901 gcacaataaa aattacacac aagaaccaag ccccaatgct gatgggcccg cctccaaaaa
     3961 ccggtttatt ctgctccctc gtcaaaagga caagaaaccg aagcaaggaa gtttaaacga
     4021 attcgggctc ggtacccggg gatcctctag agtcgacctg caggcatgca agctgatccg
     4081 gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga gcaataacta
     4141 gcataacccc ttggggcggc cgcttcgagc agacatgata agatacattg atgagtttgg
     4201 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     4261 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     4321 ttttatgttt caggttcagg gggagatgtg ggaggttttt ttaagcaagt aaaacctcta
     4381 caaatgtggt aaaatcgaat tttaacaaaa tattaacgct tacaatttcc tgatgcggta
     4441 ttttctcctt acgcatctgt gcggtatttc acaccgcata cgcggatctg cgcagcacca
     4501 tggcctgaaa taacctctga aagaggaact tggttaggta ccttctgagg cggaaagaac
     4561 cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     4621 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     4681 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     4741 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     4801 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     4861 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagcttaa ttaactgttg
     4921 acaattaatc atcggcatag tatatcggca tagtataata cgacaaggtg aggaactaaa
     4981 cccaggaggc agatcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     5041 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     5101 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     5161 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     5221 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     5281 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     5341 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     5401 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     5461 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     5521 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     5581 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     5641 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     5701 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     5761 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     5821 aagcgacgcc caacctgcca tcacgatggc cgcaataaaa tatctttatt ttcattacat
     5881 ctgtgtgttg gttttttgtg tgaatcgata gcgataagga tccgctcatg cgatgataag
     5941 ctgtcaaaca tgagaattac aacttatatc gtatggggct gacttcaggt gctacatttg
     6001 aagagataaa ttgcactgaa atctagaaat attttatctg attaataaga tgatcttctt
     6061 gagatcgttt tggtctgcgc gtaatctctt gctctgaaaa cgaaaaaacc gccttgcagg
     6121 gcggtttttc gaaggttctc tgagctacca actctttgaa ccgaggtaac tggcttggag
     6181 gagcgcagtc accaaaactt gtcctttcag tttagcctta accggcgcat gacttcaaga
     6241 ctaactcctc taaatcaatt accagtggct gctgccagtg gtgcttttgc atgtctttcc
     6301 gggttggact caagacgata gttaccggat aaggcgcagc ggtcggactg aacggggggt
     6361 tcgtgcatac agtccagctt ggagcgaact gcctacccgg aactgagtgt caggcgtgga
     6421 atgagacaaa cgcggccata acagcggaat gacaccggta aaccgaaagg caggaacagg
     6481 agagcgcacg agggagccgc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     6541 tcgccaccac tgatttgagc gtcagatttc gtgatgcttg tcaggggggc ggagcctatg
     6601 gaaaaacggc tttgccgcgg ccctctcact tccctgttaa gtatcttcct ggcatcttcc
     6661 aggaaatctc cgccccgttc gtaagccatt tccgctcgcc gcagtcgaac gaccgagcgt
     6721 agcgagtcag tgagcgagga agcggaatat atcctgtatc acatattctg ctgacgcacc
     6781 ggtgcagcct tttttctcct gccacatgaa gcacttcact gacaccctca tcagtgccaa
     6841 catagtaagc cagtatacac tccgctagcg ctgatgtccg gcggtgcttt tgccgttacg
     6901 caccaccccg tcatgagatt acgcgtttta gcggtgcgta caattaaggg attatggtaa
     6961 atccacttac tgtctgccct cgtagccatc gagataaacc gcagtactcc ggccacgatg
     7021 cgtccggcgt agaggatcga gatct
//
