LOCUS       sequence-481539-        6365 bp    DNA     circular SYN 30-JUL-2025
DEFINITION  Express NanoLuc-PDPK1 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 6365)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6365)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 30, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: NanoLuc-PDPK1 Fusion Vector
FEATURES             Location/Qualifiers
     source          1..6365
                     /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(3380..3398)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3435..3556
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3446..3465)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3500..3519
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3824..3844)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3868..4078
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        3890..4086
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3937..4072
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4101..4148
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4180..4974
                     /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(4234..4253)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4844..4863
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5038..5086
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(5325..5913)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5405..5424)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6029..6312
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
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 ggccaggacc accagccagc
     1621 tgtatgacgc cgtgcccatc cagtccagcg tggtgttatg ttcctgccca tccccatcaa
     1681 tggtgaggac ccagactgag tccagcacgc cccctggcat tcctggtggc agcaggcagg
     1741 gccccgccat ggacggcact gcagccgagc ctcggcccgg cgccggctcc ctgcagcatg
     1801 cccagcctcc gccgcagcct cggaagaagc ggcctgagga cttcaagttt gggaaaatcc
     1861 ttggggaagg ctctttttcc acggttgtcc tggctcgaga actggcaacc tccagagaat
     1921 atgcgattaa aattctggag aagcgacata tcataaaaga gaacaaggtc ccctatgtaa
     1981 ccagagagcg ggatgtcatg tcgcgcctgg atcacccctt ctttgttaag ctttacttca
     2041 catttcagga cgacgagaag ctgtatttcg gccttagtta tgccaaaaat ggagaactac
     2101 ttaaatatat tcgcaaaatc ggttcattcg atgagacctg tacccgattt tacacggctg
     2161 agattgtgtc tgctttagag tacttgcacg gcaagggcat cattcacagg gaccttaaac
     2221 cggaaaacat tttgttaaat gaagatatgc acatccagat cacagatttt ggaacagcaa
     2281 aagtcttatc cccagagagc aaacaagcca gggccaactc attcgtggga acagcgcagt
     2341 acgtttctcc agagctgctc acggagaagt ccgcctgtaa gagttcagac ctttgggctc
     2401 ttggatgcat aatataccag cttgtggcag gactcccacc attccgagct ggaaacgagt
     2461 atcttatatt tcagaagatc attaagttgg aatatgactt tccagaaaaa ttcttcccta
     2521 aggcaagaga cctcgtggag aaacttttgg ttttagatgc cacaaagcgg ttaggctgtg
     2581 aggaaatgga aggatacgga cctcttaaag cacacccgtt cttcgagtcc gtcacgtggg
     2641 agaacctgca ccagcagacg cctccgaagc tcaccgctta cctgccggct atgtcggaag
     2701 acgacgagga ctgctatggc aattatgaca atctcctgag ccagtttggc tgcatgcagg
     2761 tgtcttcgtc ctcctcctca cactccctgt cagcctccga cacgggcctg ccccagaggt
     2821 caggcagcaa catagagcag tacattcacg atctggactc gaactccttt gaactggact
     2881 tacagttttc cgaagatgag aagaggttgt tgttggagaa gcaggctggc ggaaaccctt
     2941 ggcaccagtt tgtagaaaat aatttaatac taaagatggg cccagtggat aagcggaagg
     3001 gtttatttgc aagacgacga cagctgttgc tcacagaagg accacattta tattatgtgg
     3061 atcctgtcaa caaagttctg aaaggtgaaa ttccttggtc acaagaactt cgaccagagg
     3121 ccaagaattt taaaactttc tttgtccaca cgcctaacag gacgtattat ctgatggacc
     3181 ccagcgggaa cgcacacaag tggtgcagga agatccagga ggtttggagg cagcgatacc
     3241 agagccaccc ggacgccgct gtgcaggttt aaacgaattc gggctcggta cccggggatc
     3301 ctctagagtc gacctgcagg catgcaagct gatccggctg ctaacaaagc ccgaaaggaa
     3361 gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcggccgct
     3421 tcgagcagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     3481 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     3541 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga
     3601 gatgtgggag gtttttttaa gcaagtaaaa cctctacaaa tgtggtaaaa tcgaatttta
     3661 acaaaatatt aacgcttaca atttcctgat gcggtatttt ctccttacgc atctgtgcgg
     3721 tatttcacac cgcatacgcg gatctgcgca gcaccatggc ctgaaataac ctctgaaaga
     3781 ggaacttggt taggtacctt ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta
     3841 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3901 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     3961 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     4021 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     4081 tgcaaaaagc ttaattaact gttgacaatt aatcatcggc atagtatatc ggcatagtat
     4141 aatacgacaa ggtgaggaac taaacccagg aggcagatca tgattgaaca agatggattg
     4201 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     4261 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     4321 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta
     4381 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     4441 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     4501 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     4561 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     4621 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     4681 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc
     4741 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     4801 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     4861 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     4921 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     4981 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga tggccgcaat
     5041 aaaatatctt tattttcatt acatctgtgt gttggttttt tgtgtgaatc gatagcgata
     5101 aggatcctct ttgcgcttgc gttttccctt gtccagatag cccagtagct gacattcatc
     5161 cggggtcagc accgtttctg cggactggct ttctacccgg tatcagctca ctcaaaggcg
     5221 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     5281 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     5341 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     5401 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     5461 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     5521 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     5581 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     5641 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     5701 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     5761 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     5821 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     5881 cagcagatta cgcgcagaaa aaaaggattt caagaagatc ctttgatctt ttctacgggg
     5941 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     6001 aggatcttca cctagatcct tttatagtcc ggaaatacag gaacgcacgc tggatggccc
     6061 ttcgctggga tggtgaaacc atgaaaaatg gcagcttcag tggattaagt gggggtaatg
     6121 tggcctgtac cctctggttg cataggtatt catacggtta aaatttatca ggcgcgattg
     6181 cggcagtttt tcgggtggtt tgttgccatt tttacctgtc tgctgccgtg atcgcgctga
     6241 acgcgtttta gcggtgcgta caattaaggg attatggtaa atccacttac tgtctgccct
     6301 cgtagccatc gagataaacc gcagtactcc ggccacgatg cgtccggcgt agaggatcga
     6361 gatct
//
