LOCUS       sequence-486652-        7004 bp    DNA     circular SYN 17-SEP-2025
DEFINITION  Express DGKB-NanoLuc(R) 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 7004)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7004)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 17, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: DGKb-NanoLuc Fusion Vector
FEATURES             Location/Qualifiers
     source          1..7004
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1180..1692
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     primer_bind     complement(1780..1798)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    1835..1956
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1846..1865)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1900..1919
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2201..2558
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2201..2392
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2224..2244)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2409..2544
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        2573..2620
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2652..3446
                     /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(2706..2725)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3316..3335
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3510..3558
                     /note="synthetic polyadenylation signal"
     rep_origin      3715..4260
                     /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     4161..4180
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4377..4394
                     /label=L4440
                     /note="L4440 vector, forward primer"
     enhancer        4839..5218
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5219..5422
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5373..5393
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          5558..5690
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     5673..5692
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     5734..5753
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5734..5752
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 aactctgtta ctgtagatgg acaaggcctg caggtcactc ctgtgcctgg tactcaccca
       61 cttttagttt ttgtgaaccc caaaagtggt ggaaaacaag gagaacgaat ttacagaaaa
      121 ttccagtatc tattaaatcc tcgtcaggtt tacagtcttt ctggaaatgg accaatgcca
      181 gggttaaact ttttccgtga tgttcctgac ttcagagtgt tagcctgtgg tggagatgga
      241 accgtgggct gggttttgga ttgcatagaa aaggccaatg taggcaagca tcctccagtt
      301 gcgattctgc ctcttgggac tggcaatgat ctagcaagat gcctgcgatg gggaggaggt
      361 tacgaaggtg agaatctgat gaaaattcta aaagacattg aaaacagcac agaaatcatg
      421 ttggacaggt ggaagtttga agtcatacct aatgacaaag atgagaaagg agacccagtg
      481 ccttacagta tcatcaataa ttacttttcc attggcgtgg atgcctccat tgcacacaga
      541 ttccacatca tgagagaaaa acacccagag aaattcaaca gtagaatgaa gaacaaattt
      601 tggtattttg agtttggcac atctgaaact ttctcagcca cctgcaagaa gctacatgaa
      661 tctgtagaaa tagaatgtga tggagtacag atagatttaa taaacatctc tctggaagga
      721 attgctattt tgaatatacc aagcatgcat ggaggatcca atctttgggg agagtctaag
      781 aaaagacgaa gccatcgacg aatagagaaa aaagggtctg acaaaaggac caccgtcaca
      841 gatgccaaag agttgaagtt tgcaagtcaa gatctcagtg accagctgct ggaggtggtc
      901 ggcttggaag gagccatgga gatggggcaa atatacacag gcctgaaaag tgctggccgg
      961 cggctggctc agtgctcctg cgtggtcatc aggacgagca agtctctgcc aatgcaaatt
     1021 gatggggagc catggatgca gaccccatgc acaataaaaa ttacacacaa gaaccaagcc
     1081 ccaatgctga tgggcccgcc tccaaaaacc ggtttattct gctccctcgt caaaaggaca
     1141 agaaaccgaa gcaaggaagt ttctctcggc tcgagcggcg tcttcacact cgaagatttc
     1201 gttggggact ggcgacagac agccggctac aacctggacc aagtccttga acagggaggt
     1261 gtgtccagtt tgtttcagaa tctcggggtg tccgtaactc cgatccaaag gattgtcctg
     1321 agcggtgaaa atgggctgaa gatcgacatc catgtcatca tcccgtatga aggtctgagc
     1381 ggcgaccaaa tgggccagat cgaaaaaatt tttaaggtgg tgtaccctgt ggatgatcat
     1441 cactttaagg tgatcctgca ctatggcaca ctggtaatcg acggggttac gccgaacatg
     1501 atcgactatt tcggacggcc gtatgaaggc atcgccgtgt tcgacggcaa aaagatcact
     1561 gtaacaggga ccctgtggaa cggcaacaaa attatcgacg agcgcctgat caaccccgac
     1621 ggctccctgc tgttccgagt aaccatcaac ggagtgaccg gctggcggct gtgcgaacgc
     1681 attctggcgt aaggccgcga ctctagagtc gacctgcagg catgcaagct gatccggctg
     1741 ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagcat
     1801 aaccccttgg ggcggccgct tcgagcagac atgataagat acattgatga gtttggacaa
     1861 accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct
     1921 ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt
     1981 atgtttcagg ttcaggggga gatgtgggag gtttttttaa gcaagtaaaa cctctacaaa
     2041 tgtggtaaaa tcgaatttta acaaaatatt aacgcttaca atttcctgat gcggtatttt
     2101 ctccttacgc atctgtgcgg tatttcacac cgcatacgcg gatctgcgca gcaccatggc
     2161 ctgaaataac ctctgaaaga ggaacttggt taggtacctt ctgaggcgga aagaaccagc
     2221 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     2281 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     2341 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     2401 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     2461 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     2521 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gcttaattaa ctgttgacaa
     2581 ttaatcatcg gcatagtata tcggcatagt ataatacgac aaggtgagga actaaaccca
     2641 ggaggcagat catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     2701 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     2761 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     2821 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     2881 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     2941 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     3001 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     3061 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     3121 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     3181 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     3241 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     3301 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     3361 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     3421 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     3481 gacgcccaac ctgccatcac gatggccgca ataaaatatc tttattttca ttacatctgt
     3541 gtgttggttt tttgtgtgaa tcgatagcga taaggatccg ctcatgcgat gataagctgt
     3601 caaacatgag aattacaact tatatcgtat ggggctgact tcaggtgcta catttgaaga
     3661 gataaattgc actgaaatct agaaatattt tatctgatta ataagatgat cttcttgaga
     3721 tcgttttggt ctgcgcgtaa tctcttgctc tgaaaacgaa aaaaccgcct tgcagggcgg
     3781 tttttcgaag gttctctgag ctaccaactc tttgaaccga ggtaactggc ttggaggagc
     3841 gcagtcacca aaacttgtcc tttcagttta gccttaaccg gcgcatgact tcaagactaa
     3901 ctcctctaaa tcaattacca gtggctgctg ccagtggtgc ttttgcatgt ctttccgggt
     3961 tggactcaag acgatagtta ccggataagg cgcagcggtc ggactgaacg gggggttcgt
     4021 gcatacagtc cagcttggag cgaactgcct acccggaact gagtgtcagg cgtggaatga
     4081 gacaaacgcg gccataacag cggaatgaca ccggtaaacc gaaaggcagg aacaggagag
     4141 cgcacgaggg agccgccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     4201 caccactgat ttgagcgtca gatttcgtga tgcttgtcag gggggcggag cctatggaaa
     4261 aacggctttg ccgcggccct ctcacttccc tgttaagtat cttcctggca tcttccagga
     4321 aatctccgcc ccgttcgtaa gccatttccg ctcgccgcag tcgaacgacc gagcgtagcg
     4381 agtcagtgag cgaggaagcg gaatatatcc tgtatcacat attctgctga cgcaccggtg
     4441 cagccttttt tctcctgcca catgaagcac ttcactgaca ccctcatcag tgccaacata
     4501 gtaagccagt atacactccg ctagcgctga tgtccggcgg tgcttttgcc gttacgcacc
     4561 accccgtcat gagattacgc gttttagcgg tgcgtacaat taagggatta tggtaaatcc
     4621 acttactgtc tgccctcgta gccatcgaga taaaccgcag tactccggcc acgatgcgtc
     4681 cggcgtagag gatcgagatc ttcaatattg gccattagcc atattattca ttggttatat
     4741 agcataaatc aatattggct attggccatt gcatacgttg tatctatatc ataatatgta
     4801 catttatatt ggctcatgtc caatatgacc gccatgttgg cattgattat tgactagtta
     4861 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
     4921 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
     4981 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
     5041 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtcc
     5101 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
     5161 cttacgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
     5221 gatgcggttt tggcagtaca ccaatgggcg tggatagcgg tttgactcac ggggatttcc
     5281 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
     5341 tccaaaatgt cgtaataacc ccgccccgtt gacgcaaatg ggcggtaggc gtgtacggtg
     5401 ggaggtctat ataagcagag ctggtttagt gaaccgtcag atcactagaa gctttattgc
     5461 ggtagtttat cacagttaaa ttgctaacgc agtcagtgct tctgacacaa cagtctcgaa
     5521 cttaagctgc agaagttggt cgtgaggcac tgggcaggta agtatcaagg ttacaagaca
     5581 ggtttaagga gaccaataga aactgggctt gtcgagacag agaagactct tgcgtttctg
     5641 ataggcacct attggtctta ctgacatcca ctttgccttt ctctccacag gtgtccactc
     5701 ccagttcaat tacagctctt aaggctagag tattaatacg actcactata gggctagcga
     5761 tcgccatgac aaaccaggaa aaatgggccc acctcagccc ttcggaattt tcccaacttc
     5821 agaaatatgc tgagtattct acaaagaaat taaaggatgt tcttgaagaa ttccatggta
     5881 atggtgtgct tgcaaagtat aatcctgaag ggaaacaaga cattcttaac caaacaatag
     5941 attttgaagg tttcaaacta ttcatgaaga cattcctgga agccgagctt cctgatgatt
     6001 tcactgcaca ccttttcatg tcatttagca acaagtttcc tcattctagt ccaatggtaa
     6061 aaagtaagcc tgctctccta tcaggcggtc tgagaatgaa taaaggtgcc atcacccctc
     6121 cccggactac ttctcctgca aatacgtgtt ccccagaagt aatccatctg aaggacattg
     6181 tctgttacct gtctctgctt gaaagaggaa gacctgagga taagcttgag tttatgtttc
     6241 gcctttatga cacggatggg aatggcttcc tggacagctc ggagctagaa aatatcatca
     6301 gtcagatgat gcatgttgca gaataccttg agtgggatgt cactgaactt aatccaatcc
     6361 tccatgaaat gatggaagaa attgactatg atcatgatgg aaccgtgtct caaggaggaa
     6421 tgacaacgat tccacttctt gtgctcctgg gcttagaaaa taacgtgaag gatgatggac
     6481 agcacgtgtg gcgactgaag cactttaaca aacctgccta ttgcaacctt tgcctgaaca
     6541 tgctgattgg cgtggggaag cagggcctct gctgttcctt ctgcaagtac acagtccatg
     6601 agcgctgtgt ggctcgagca cctccctctt gcatcaagac ctatgtgaag tccaaaagga
     6661 acactgatgt catgcaccat tactgggttg aaggtaactg cccaaccaag tgtgataagt
     6721 gccacaaaac tgttaaatgt taccagggcc tgacaggact gcattgtgtt tggtgtcaga
     6781 tcacactgca taataaatgt gcttctcatc taaaacctga atgtgactgt ggacctttga
     6841 aggaccatat tttaccaccc acaacaatct gtccagtggt actgcagact ctgcccactt
     6901 caggagtttc agttcctgag gaaagacaat caacagtgaa aaaggaaaag agtggttccc
     6961 agcagccaaa caaagtgatt gacaagaata aaatgcaaag agcc
//
