LOCUS       sequence-474698-        5387 bp    DNA     circular SYN 05-JUN-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 5387)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 5387)
  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: DGKb-NanoLuc Fusion Vector
FEATURES             Location/Qualifiers
     source          1..5387
                     /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             1866..2378
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     primer_bind     complement(2466..2484)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2521..2642
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2532..2551)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2586..2605
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2887..3244
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2887..3078
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2910..2930)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3095..3230
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3259..3306
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3338..4132
                     /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(3392..3411)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4002..4021
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4196..4244
                     /note="synthetic polyadenylation signal"
     rep_origin      4401..4946
                     /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     4847..4866
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5063..5080
                     /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 ggctagcgat cgccatgtcc catcactggg
     1081 ggtacggcaa acacaacgga cctgagcact ggcataagga cttccccatt gccaagggag
     1141 agcgccagtc ccctgttgac atcgacactc atacagccaa gtatgaccct tccctgaagc
     1201 ccctgtctgt ttcctatgat caagcaactt ccctgaggat cctcaacaat ggtcatgctt
     1261 tcaacgtgga gtttgatgac tctcaggaca aagcagtgct caagggagga cccctggatg
     1321 gcacttacag attgattcag tttcactttc actggggttc acttgatgga caaggttcag
     1381 agcatactgt ggataaaaag aaatatgctg cagaacttca cttggttcac tggaacacca
     1441 aatatgggga ttttgggaaa gctgtgcagc aacctgatgg actggccgtt ctaggtattt
     1501 ttttgaaggt tggcagcgct aaaccgggcc ttcagaaagt tgttgatgtg ctggattcca
     1561 ttaaaacaaa gggcaagagt gctgacttca ctaacttcga tcctcgtggc ctccttcctg
     1621 aatccttgga ttactggacc tacccaggct cactgaccac ccctcctctt ctggaatgtg
     1681 tgacctggat tgtgctcaag gaacccatca gcgtcagcag cgagcaggtg ttgaaattcc
     1741 gtaaacttaa cttcaatggg gagggtgaac ccgaagaact gatggtggac aactggcgcc
     1801 cagctcagcc actgaagaac aggcaaatca aagcttcctt caaagtttct ctcggctcga
     1861 gcggcgtctt cacactcgaa gatttcgttg gggactggcg acagacagcc ggctacaacc
     1921 tggaccaagt ccttgaacag ggaggtgtgt ccagtttgtt tcagaatctc ggggtgtccg
     1981 taactccgat ccaaaggatt gtcctgagcg gtgaaaatgg gctgaagatc gacatccatg
     2041 tcatcatccc gtatgaaggt ctgagcggcg accaaatggg ccagatcgaa aaaattttta
     2101 aggtggtgta ccctgtggat gatcatcact ttaaggtgat cctgcactat ggcacactgg
     2161 taatcgacgg ggttacgccg aacatgatcg actatttcgg acggccgtat gaaggcatcg
     2221 ccgtgttcga cggcaaaaag atcactgtaa cagggaccct gtggaacggc aacaaaatta
     2281 tcgacgagcg cctgatcaac cccgacggct ccctgctgtt ccgagtaacc atcaacggag
     2341 tgaccggctg gcggctgtgc gaacgcattc tggcgtaagg ccgcgactct agagtcgacc
     2401 tgcaggcatg caagctgatc cggctgctaa caaagcccga aaggaagctg agttggctgc
     2461 tgccaccgct gagcaataac tagcataacc ccttggggcg gccgcttcga gcagacatga
     2521 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     2581 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     2641 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggagatg tgggaggttt
     2701 ttttaagcaa gtaaaacctc tacaaatgtg gtaaaatcga attttaacaa aatattaacg
     2761 cttacaattt cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     2821 tacgcggatc tgcgcagcac catggcctga aataacctct gaaagaggaa cttggttagg
     2881 taccttctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc
     2941 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     3001 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     3061 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     3121 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     3181 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     3241 caaaaagctt aattaactgt tgacaattaa tcatcggcat agtatatcgg catagtataa
     3301 tacgacaagg tgaggaacta aacccaggag gcagatcatg attgaacaag atggattgca
     3361 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     3421 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     3481 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
     3541 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     3601 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     3661 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     3721 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     3781 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     3841 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
     3901 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     3961 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     4021 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     4081 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     4141 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgatg gccgcaataa
     4201 aatatcttta ttttcattac atctgtgtgt tggttttttg tgtgaatcga tagcgataag
     4261 gatccgctca tgcgatgata agctgtcaaa catgagaatt acaacttata tcgtatgggg
     4321 ctgacttcag gtgctacatt tgaagagata aattgcactg aaatctagaa atattttatc
     4381 tgattaataa gatgatcttc ttgagatcgt tttggtctgc gcgtaatctc ttgctctgaa
     4441 aacgaaaaaa ccgccttgca gggcggtttt tcgaaggttc tctgagctac caactctttg
     4501 aaccgaggta actggcttgg aggagcgcag tcaccaaaac ttgtcctttc agtttagcct
     4561 taaccggcgc atgacttcaa gactaactcc tctaaatcaa ttaccagtgg ctgctgccag
     4621 tggtgctttt gcatgtcttt ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     4681 gcggtcggac tgaacggggg gttcgtgcat acagtccagc ttggagcgaa ctgcctaccc
     4741 ggaactgagt gtcaggcgtg gaatgagaca aacgcggcca taacagcgga atgacaccgg
     4801 taaaccgaaa ggcaggaaca ggagagcgca cgagggagcc gccaggggga aacgcctggt
     4861 atctttatag tcctgtcggg tttcgccacc actgatttga gcgtcagatt tcgtgatgct
     4921 tgtcaggggg gcggagccta tggaaaaacg gctttgccgc ggccctctca cttccctgtt
     4981 aagtatcttc ctggcatctt ccaggaaatc tccgccccgt tcgtaagcca tttccgctcg
     5041 ccgcagtcga acgaccgagc gtagcgagtc agtgagcgag gaagcggaat atatcctgta
     5101 tcacatattc tgctgacgca ccggtgcagc cttttttctc ctgccacatg aagcacttca
     5161 ctgacaccct catcagtgcc aacatagtaa gccagtatac actccgctag cgctgatgtc
     5221 cggcggtgct tttgccgtta cgcaccaccc cgtcatgaga ttacgcgttt tagcggtgcg
     5281 tacaattaag ggattatggt aaatccactt actgtctgcc ctcgtagcca tcgagataaa
     5341 ccgcagtact ccggccacga tgcgtccggc gtagaggatc gagatct
//
