LOCUS       sequence-470775-        7838 bp    DNA     circular SYN 21-MAY-2025
DEFINITION  Express NanoLuc(R)-DGKI 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 7838)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 7838)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 21, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: NanoLuc-DGKi Fusion Vector
FEATURES             Location/Qualifiers
     source          1..7838
                     /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(4907..4925)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    4962..5083
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4973..4992)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5027..5046
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        5328..5685
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5328..5519
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5351..5371)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5536..5671
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        5700..5747
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5779..6573
                     /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(5833..5852)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6443..6462
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6647..6695
                     /note="synthetic polyadenylation signal"
     rep_origin      6852..7397
                     /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     7298..7317
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7514..7531
                     /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 ggatgctgct ggaagaggtt
     1621 gtcatttact acctctacca gcggctagag gacctgctag ggctcctgca gctgcagctg
     1681 ctgcagcagc aagtcctcca ggtccatgta gtggagctgc atgtgctcca agtgcagcag
     1741 ctggagcagg tgctatgaat cctagttcat cggcgggaga ggagaaaggg gcgacgggcg
     1801 gcagcagcag cagcggaagc ggtgctggta gttgttgtct aggagcagaa ggtggtgcgg
     1861 acccgcgggg cgcagggtca gccgctgcag caggagctgc tgcactggac gaacctgctg
     1921 cagcaggtca gaaggagaag gacgaagcgc tggaggagaa gctgaggaac ttaactttcc
     1981 ggaagcaggt ctcgtacagg aaagcaatct cccgggcagg cctccagcat ctggctcctg
     2041 cacatcccct cagccttcct gtggcaaatg gtccagccaa ggagcccaga gcgactttgg
     2101 actggagtga gaatgccgtg aatggagaac acctgtggct ggagaccaac gtctcgggag
     2161 acctctgcta ccttggagag gagaactgcc aagtcagatt tgcaaaatca gctctcagga
     2221 ggaagtgtgc agtctgtaaa atcgtcgtcc acaccgcctg cattgagcag ctagaaaaga
     2281 ttaatttcag atgtaaacca acatttcgag aaggaggctc aaggtcacca agagaaaatt
     2341 ttgtacgtca tcactgggtg cacaggcgtc ggcaggaggg gaaatgtaag cagtgtggta
     2401 agggcttcca gcaaaagttc tccttccaca gtaaagagat tgtggctatc agctgttcct
     2461 ggtgcaagca ggcgtttcac aataaggtga cctgcttcat gctgcatcac attgaagaac
     2521 cctgctccct gggggctcat gctgctgtta ttgtcccgcc cacttggatc attaaggtga
     2581 agaaacctca gaactccctg aaggcttcaa atcggaagaa gaagagaaca agctttaaaa
     2641 gaaaagccag taaaagaggg atggaacagg aaaacaaagg tcgtcctttt gtgataaaac
     2701 ccatctcttc tcctctcatg aaacccttgc ttgtatttgt gaatcccaag agtggaggca
     2761 accagggaac caaagtcctg cagatgttca tgtggtacct gaatccacgg caagtctttg
     2821 atctttctca ggaagggcca aaagatgcgc ttgaattgta taggaaagta ccaaatctgc
     2881 gaattctggc ctgtggtggg gatggaacgg tgggctggat cctttccatc ctggatgaac
     2941 tgcagctgag ccctcagcct cctgtggggg tccttcctct ggggactggg aatgacctgg
     3001 ctcgaactct caactgggga gggggctaca ctgatgaacc tgtttctaaa attctttgtc
     3061 aagttgaaga tggtacagta gtacaactag atcgttggaa tctccatgtg gaaagaaacc
     3121 ccgacttgcc tccagaagaa cttgaagatg gcgtatgtaa gctccctctg aatgttttca
     3181 ataactactt cagccttgga tttgatgccc atgtcacact ggagttccat gaatccagag
     3241 aagcaaatcc agagaaattc aacagtcgtt ttcgaaataa aatgttctat gcaggggcag
     3301 ctttttctga cttcctacag agaagttcta gagatctatc caaacatgtt aaagttgttt
     3361 gtgatggaac agatctcacc ccaaagattc aggaactgaa gttccagtgt atagtatttt
     3421 taaatatacc cagatattgt gctggcacaa tgccctgggg aaacccaggt gatcaccatg
     3481 atttcgaacc tcagcgtcat gatgatggtt atattgaagt cattggattt accatggcct
     3541 ctttggcagc cctgcaagtt gggggccatg gagagaggct acaccagtgt cgagaagtca
     3601 tgcttctaac ttacaaatcc atccccatgc aagtggatgg ggagccctgt aggttggccc
     3661 cagctatgat tcggatctcc ctgaggaatc aggccaacat ggtacagaag agcaagagga
     3721 gaacatccat gcctttactc aatgatcccc agtctgtccc agatcgtctg aggatccggg
     3781 tgaacaaaat cagtttacaa gactatgaag gattccacta tgacaaggag aaactccgag
     3841 aagcttctat ttcagactgg ttaagaacca ttgctgggga actagtgcag tcatttggag
     3901 cgatacctct gggtattcta gttgtgcgtg gagactgtga tttggagact tgccgtatgt
     3961 acatagaccg cctacaggag gacctacagt cagtttcttc tggctcccag agagttcatt
     4021 accaggacca tgaaacctcc ttccccaggg ctctctcagc acagaggctc tctcctcggt
     4081 ggtgcttcct agatgacaga tctcaggaac atttgcactt tgtgatggag atttcccaag
     4141 atgagatttt tattctggac ccagatatgg tggtgtcaca gccggcgggg acacctccgg
     4201 gcatgcctga cctggtggtg gaacaagcct cggggatctc agactggtgg aatcctgccc
     4261 tgcggaaacg catgctgagt gacagtgggc tggggatgat agctccctat tatgaggact
     4321 cagatctgaa agatctcagc cactcccgcg tgctacagtc accagtctct tcagaagatc
     4381 atgcaatttt gcaggcagta atagctggtg atcttatgaa gctaatagaa agctataaaa
     4441 atggaggcag tctgctaatt cagggaccag accactgttc actccttcac tacgcagcta
     4501 aaaccggcaa cggggagatt gtgaaatata tccttgacca cggaccttcc gagttattgg
     4561 atatggcaga cagtgaaacg ggtgagactg cactgcacaa ggctgcctgc cagcggaacc
     4621 gggctgtgtg ccagcttctg gtggatgcag gagcatctct gagaaagacg gactccaagg
     4681 gtaagacacc tcaagaaaga gcacagcagg ctggggaccc agacttggct gcttacctag
     4741 aaagccgtca gaactataag gtcattggcc atgaggacct ggaaactgct gttgtttaaa
     4801 cgaattcggg ctcggtaccc ggggatcctc tagagtcgac ctgcaggcat gcaagctgat
     4861 ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc tgagcaataa
     4921 ctagcataac cccttggggc ggccgcttcg agcagacatg ataagataca ttgatgagtt
     4981 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     5041 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     5101 tcattttatg tttcaggttc agggggagat gtgggaggtt tttttaagca agtaaaacct
     5161 ctacaaatgt ggtaaaatcg aattttaaca aaatattaac gcttacaatt tcctgatgcg
     5221 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgcggat ctgcgcagca
     5281 ccatggcctg aaataacctc tgaaagagga acttggttag gtaccttctg aggcggaaag
     5341 aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     5401 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     5461 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     5521 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     5581 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     5641 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct taattaactg
     5701 ttgacaatta atcatcggca tagtatatcg gcatagtata atacgacaag gtgaggaact
     5761 aaacccagga ggcagatcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     5821 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     5881 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     5941 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
     6001 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     6061 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     6121 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     6181 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     6241 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     6301 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     6361 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     6421 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     6481 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     6541 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     6601 accaagcgac gcccaacctg ccatcacgat ggccgdgkgt nrtnmcaata aaatatcttt
     6661 attttcatta catctgtgtg ttggtttttt gtgtgaatcg atagcgataa ggatccgctc
     6721 atgcgatgat aagctgtcaa acatgagaat tacaacttat atcgtatggg gctgacttca
     6781 ggtgctacat ttgaagagat aaattgcact gaaatctaga aatattttat ctgattaata
     6841 agatgatctt cttgagatcg ttttggtctg cgcgtaatct cttgctctga aaacgaaaaa
     6901 accgccttgc agggcggttt ttcgaaggtt ctctgagcta ccaactcttt gaaccgaggt
     6961 aactggcttg gaggagcgca gtcaccaaaa cttgtccttt cagtttagcc ttaaccggcg
     7021 catgacttca agactaactc ctctaaatca attaccagtg gctgctgcca gtggtgcttt
     7081 tgcatgtctt tccgggttgg actcaagacg atagttaccg gataaggcgc agcggtcgga
     7141 ctgaacgggg ggttcgtgca tacagtccag cttggagcga actgcctacc cggaactgag
     7201 tgtcaggcgt ggaatgagac aaacgcggcc ataacagcgg aatgacaccg gtaaaccgaa
     7261 aggcaggaac aggagagcgc acgagggagc cgccaggggg aaacgcctgg tatctttata
     7321 gtcctgtcgg gtttcgccac cactgatttg agcgtcagat ttcgtgatgc ttgtcagggg
     7381 ggcggagcct atggaaaaac ggctttgccg cggccctctc acttccctgt taagtatctt
     7441 cctggcatct tccaggaaat ctccgccccg ttcgtaagcc atttccgctc gccgcagtcg
     7501 aacgaccgag cgtagcgagt cagtgagcga ggaagcggaa tatatcctgt atcacatatt
     7561 ctgctgacgc accggtgcag ccttttttct cctgccacat gaagcacttc actgacaccc
     7621 tcatcagtgc caacatagta agccagtata cactccgcta gcgctgatgt ccggcggtgc
     7681 ttttgccgtt acgcaccacc ccgtcatgag attacgcgtt ttagcggtgc gtacaattaa
     7741 gggattatgg taaatccact tactgtctgc cctcgtagcc atcgagataa accgcagtac
     7801 tccggccacg atgcgtccgg cgtagaggat cgagatct
//
