LOCUS       sequence-479783-        6448 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Express NanoLuc-PPM1D 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 6448)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6448)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 7, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: NanoLuc-PPM1D Fusion Vector
FEATURES             Location/Qualifiers
     source          1..6448
                     /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(3527..3545)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3582..3703
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3593..3612)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3647..3666
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3948..4305
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3948..4139
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3971..3991)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4156..4291
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4320..4367
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4399..5193
                     /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(4453..4472)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5063..5082
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5257..5305
                     /note="synthetic polyadenylation signal"
     rep_origin      5462..6007
                     /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     5908..5927
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6124..6141
                     /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 ggcggggctg tactcgctgg
     1621 gagtgagcgt cttctccgac cagggcggga ggaagtacat ggaggacgtt actcaaatcg
     1681 ttgtggagcc cgaaccgacg gctgaagaaa agccctcgcc gcggcggtcg ctgtctcagc
     1741 cgttgcctcc gcggccgtcg ccggccgccc ttcccggcgg cgaagtctcg gggaaaggcc
     1801 cagcggtggc agcccgagag gctcgcgacc ctctcccgga cgccggggcc tcgccggcac
     1861 ctagccgctg ctgccgccgc cgttcctccg tggccttttt cgccgtgtgc gacgggcacg
     1921 gcgggcggga ggcggcacag tttgcccggg agcacttgtg gggtttcatc aagaagcaga
     1981 agggtttcac ctcgtccgag ccggctaagg tttgcgctgc catccgcaaa ggctttctcg
     2041 cttgtcacct tgccatgtgg aagaaactgg cggaatggcc aaagactatg acgggtcttc
     2101 ctagcacatc agggacaact gccagtgtgg tcatcattcg gggcatgaag atgtatgtag
     2161 ctcacgtagg tgactcaggg gtggttcttg gaattcagga tgacccgaag gatgactttg
     2221 tcagagctgt ggaggtgaca caggaccata agccagaact tcccaaggaa agagaacgaa
     2281 tcgaaggact tggtgggagt gtaatgaaca agtctggggt gaatcgtgta gtttggaaac
     2341 gacctcgact cactcacaat ggacctgtta gaaggagcac agttattgac cagattcctt
     2401 ttctggcagt agcaagagca cttggtgatt tgtggagcta tgatttcttc agtggtgaat
     2461 ttgtggtgtc acctgaacca gacacaagtg tccacactct tgaccctcag aagcacaagt
     2521 atattatatt ggggagtgat ggactttgga atatgattcc accacaagat gccatctcaa
     2581 tgtgccagga ccaagaggag aaaaaatacc tgatgggtga gcatggacaa tcttgtgcca
     2641 aaatgcttgt gaatcgagca ttgggccgct ggaggcagcg tatgctccga gcagataaca
     2701 ctagtgccat agtaatctgc atctctccag aagtggacaa tcagggaaac tttaccaatg
     2761 aagatgagtt atacctgaac ctgactgaca gcccttccta taatagtcaa gaaacctgtg
     2821 tgatgactcc ttccccatgt tctacaccac cagtcaagtc actggaggag gatccatggc
     2881 caagggtgaa ttctaaggac catatacctg ccctggttcg tagcaatgcc ttctcagaga
     2941 attttttaga ggtttcagct gagatagctc gagagaatgt ccaaggtgta gtcataccct
     3001 caaaagatcc agaaccactt gaagaaaatt gcgctaaagc cctgacttta aggatacatg
     3061 attctttgaa taatagcctt ccaattggcc ttgtgcctac taattcaaca aacactgtca
     3121 tggaccaaaa aaatttgaag atgtcaactc ctggccaaat gaaagcccaa gaaattgaaa
     3181 gaacccctcc aacaaacttt aaaaggacat tagaagagtc caattctggc cccctgatga
     3241 agaagcatag acgaaatggc ttaagtcgaa gtagtggtgc tcagcctgca agtctcccca
     3301 caacctcaca gcgaaagaac tctgttaaac tcaccatgcg acgcagactt aggggccaga
     3361 agaaaattgg aaatccttta cttcatcaac acaggaaaac tgtttgtgtt tgcgtttaaa
     3421 cgaattcggg ctcggtaccc ggggatcctc tagagtcgac ctgcaggcat gcaagctgat
     3481 ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc tgagcaataa
     3541 ctagcataac cccttggggc ggccgcttcg agcagacatg ataagataca ttgatgagtt
     3601 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     3661 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     3721 tcattttatg tttcaggttc agggggagat gtgggaggtt tttttaagca agtaaaacct
     3781 ctacaaatgt ggtaaaatcg aattttaaca aaatattaac gcttacaatt tcctgatgcg
     3841 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgcggat ctgcgcagca
     3901 ccatggcctg aaataacctc tgaaagagga acttggttag gtaccttctg aggcggaaag
     3961 aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     4021 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     4081 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     4141 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     4201 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     4261 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct taattaactg
     4321 ttgacaatta atcatcggca tagtatatcg gcatagtata atacgacaag gtgaggaact
     4381 aaacccagga ggcagatcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     4441 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     4501 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     4561 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
     4621 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     4681 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     4741 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     4801 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     4861 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     4921 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     4981 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     5041 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     5101 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     5161 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     5221 accaagcgac gcccaacctg ccatcacgat ggccgcaata aaatatcttt attttcatta
     5281 catctgtgtg ttggtttttt gtgtgaatcg atagcgataa ggatccgctc atgcgatgat
     5341 aagctgtcaa acatgagaat tacaacttat atcgtatggg gctgacttca ggtgctacat
     5401 ttgaagagat aaattgcact gaaatctaga aatattttat ctgattaata agatgatctt
     5461 cttgagatcg ttttggtctg cgcgtaatct cttgctctga aaacgaaaaa accgccttgc
     5521 agggcggttt ttcgaaggtt ctctgagcta ccaactcttt gaaccgaggt aactggcttg
     5581 gaggagcgca gtcaccaaaa cttgtccttt cagtttagcc ttaaccggcg catgacttca
     5641 agactaactc ctctaaatca attaccagtg gctgctgcca gtggtgcttt tgcatgtctt
     5701 tccgggttgg actcaagacg atagttaccg gataaggcgc agcggtcgga ctgaacgggg
     5761 ggttcgtgca tacagtccag cttggagcga actgcctacc cggaactgag tgtcaggcgt
     5821 ggaatgagac aaacgcggcc ataacagcgg aatgacaccg gtaaaccgaa aggcaggaac
     5881 aggagagcgc acgagggagc cgccaggggg aaacgcctgg tatctttata gtcctgtcgg
     5941 gtttcgccac cactgatttg agcgtcagat ttcgtgatgc ttgtcagggg ggcggagcct
     6001 atggaaaaac ggctttgccg cggccctctc acttccctgt taagtatctt cctggcatct
     6061 tccaggaaat ctccgccccg ttcgtaagcc atttccgctc gccgcagtcg aacgaccgag
     6121 cgtagcgagt cagtgagcga ggaagcggaa tatatcctgt atcacatatt ctgctgacgc
     6181 accggtgcag ccttttttct cctgccacat gaagcacttc actgacaccc tcatcagtgc
     6241 caacatagta agccagtata cactccgcta gcgctgatgt ccggcggtgc ttttgccgtt
     6301 acgcaccacc ccgtcatgag attacgcgtt ttagcggtgc gtacaattaa gggattatgg
     6361 taaatccact tactgtctgc cctcgtagcc atcgagataa accgcagtac tccggccacg
     6421 atgcgtccgg cgtagaggat cgagatct
//
