LOCUS       sequence-475296-        6437 bp    DNA     circular SYN 25-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 6437)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6437)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 25, 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            "Unpublished"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..6437
                     /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"
     promoter        3801..4158
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3801..3992
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3824..3844)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4009..4144
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4173..4220
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4252..5046
                     /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(4306..4325)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4916..4935
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5110..5158
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(5397..5985)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5477..5496)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6101..6384
                     /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 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3961 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     4021 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     4081 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     4141 ggcctaggct tttgcaaaaa gcttaattaa ctgttgacaa ttaatcatcg gcatagtata
     4201 tcggcatagt ataatacgac aaggtgagga actaaaccca ggaggcagat catgattgaa
     4261 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     4321 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     4381 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     4441 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     4501 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     4561 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     4621 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     4681 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     4741 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     4801 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     4861 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     4921 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     4981 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     5041 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     5101 gatggccgca ataaaatatc tttattttca ttacatctgt gtgttggttt tttgtgtgaa
     5161 tcgatagcga taaggatcct ctttgcgctt gcgttttccc ttgtccagat agcccagtag
     5221 ctgacattca tccggggtca gcaccgtttc tgcggactgg ctttctaccc ggtatcagct
     5281 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     5341 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     5401 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     5461 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     5521 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     5581 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     5641 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     5701 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     5761 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     5821 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     5881 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     5941 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ttcaagaaga tcctttgatc
     6001 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     6061 agattatcaa aaaggatctt cacctagatc cttttatagt ccggaaatac aggaacgcac
     6121 gctggatggc ccttcgctgg gatggtgaaa ccatgaaaaa tggcagcttc agtggattaa
     6181 gtgggggtaa tgtggcctgt accctctggt tgcataggta ttcatacggt taaaatttat
     6241 caggcgcgat tgcggcagtt tttcgggtgg tttgttgcca tttttacctg tctgctgccg
     6301 tgatcgcgct gaacgcgttt tagcggtgcg tacaattaag ggattatggt aaatccactt
     6361 actgtctgcc ctcgtagcca tcgagataaa ccgcagtact ccggccacga tgcgtccggc
     6421 gtagaggatc gagatct
//
