LOCUS       sequence-471677-        5431 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express NanoLuc(R)-15-PGDH 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 5431)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5431)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: NanoLuc-15-PGDH Fusion Vector
FEATURES             Location/Qualifiers
     source          1..5431
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(283..302)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(284..302)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(344..363)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     intron          346..478
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        complement(614..817)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(643..663)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        818..1197
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(1642..1659)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1776..2321)
                     /direction=LEFT
                     /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     complement(1856..1875)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    2478..2526
                     /note="synthetic polyadenylation signal"
     CDS             complement(2590..3384)
                     /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(2701..2720)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3311..3330
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(3416..3463)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(3478..3835)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3492..3627
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        3644..3835
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     3792..3812
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    4080..4201
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4117..4136)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4171..4190
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4238..4256
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(join(5189..5431,1..270))
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
ORIGIN
        1 aaagtgatga tcatccacag ggtacaccac cttaaaaatt ttttcgatct ggcccatttg
       61 gtcgccgctc agaccttcat acgggatgat gacatggatg tcgatcttca gcccattttc
      121 accgctcagg acaatccttt ggatcggagt tacggacacc ccgagattct gaaacaaact
      181 ggacacacct ccctgttcaa ggacttggtc caggttgtag ccggctgtct gtcgccagtc
      241 cccaacgaaa tcttcgagtg tgaagaccat ggtgagcgct agccctatag tgagtcgtat
      301 taatactcta gccttaagag ctgtaattga actgggagtg gacacctgtg gagagaaagg
      361 caaagtggat gtcagtaaga ccaataggtg cctatcagaa acgcaagagt cttctctgtc
      421 tcgacaagcc cagtttctat tggtctcctt aaacctgtct tgtaaccttg atacttacct
      481 gcccagtgcc tcacgaccaa cttctgcagc ttaagttcga gactgttgtg tcagaagcac
      541 tgactgcgtt agcaatttaa ctgtgataaa ctaccgcaat aaagcttcta gtgatctgac
      601 ggttcactaa accagctctg cttatataga cctcccaccg tacacgccta ccgcccattt
      661 gcgtcaacgg ggcggggtta ttacgacatt ttggaaagtc ccgttgattt tggtgccaaa
      721 acaaactccc attgacgtca atggggtgga gacttggaaa tccccgtgag tcaaaccgct
      781 atccacgccc attggtgtac tgccaaaacc gcatcaccat ggtaatagcg atgactaata
      841 cgtagatgta ctgccaagta ggaaagtccc gtaaggtcat gtactgggca taatgccagg
      901 cgggccattt accgtcattg acgtcaatag ggggcggact tggcatatga tacacttgat
      961 gtactgccaa gtgggcagtt taccgtaaat actccaccca ttgacgtcaa tggaaagtcc
     1021 ctattggcgt tactatggga acatacgtca ttattgacgt caatgggcgg gggtcgttgg
     1081 gcggtcagcc aggcgggcca tttaccgtaa gttatgtaac gcggaactcc atatatgggc
     1141 tatgaactaa tgaccccgta attgattact attaataact agtcaataat caatgccaac
     1201 atggcggtca tattggacat gagccaatat aaatgtacat attatgatat agatacaacg
     1261 tatgcaatgg ccaatagcca atattgattt atgctatata accaatgaat aatatggcta
     1321 atggccaata ttgaagatct cgatcctcta cgccggacgc atcgtggccg gagtactgcg
     1381 gtttatctcg atggctacga gggcagacag taagtggatt taccataatc ccttaattgt
     1441 acgcaccgct aaaacgcgta atctcatgac ggggtggtgc gtaacggcaa aagcaccgcc
     1501 ggacatcagc gctagcggag tgtatactgg cttactatgt tggcactgat gagggtgtca
     1561 gtgaagtgct tcatgtggca ggagaaaaaa ggctgcaccg gtgcgtcagc agaatatgtg
     1621 atacaggata tattccgctt cctcgctcac tgactcgcta cgctcggtcg ttcgactgcg
     1681 gcgagcggaa atggcttacg aacggggcgg agatttcctg gaagatgcca ggaagatact
     1741 taacagggaa gtgagagggc cgcggcaaag ccgtttttcc ataggctccg cccccctgac
     1801 aagcatcacg aaatctgacg ctcaaatcag tggtggcgaa acccgacagg actataaaga
     1861 taccaggcgt ttccccctgg cggctccctc gtgcgctctc ctgttcctgc ctttcggttt
     1921 accggtgtca ttccgctgtt atggccgcgt ttgtctcatt ccacgcctga cactcagttc
     1981 cgggtaggca gttcgctcca agctggactg tatgcacgaa ccccccgttc agtccgaccg
     2041 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gaaagacatg caaaagcacc
     2101 actggcagca gccactggta attgatttag aggagttagt cttgaagtca tgcgccggtt
     2161 aaggctaaac tgaaaggaca agttttggtg actgcgctcc tccaagccag ttacctcggt
     2221 tcaaagagtt ggtagctcag agaaccttcg aaaaaccgcc ctgcaaggcg gttttttcgt
     2281 tttcagagca agagattacg cgcagaccaa aacgatctca agaagatcat cttattaatc
     2341 agataaaata tttctagatt tcagtgcaat ttatctcttc aaatgtagca cctgaagtca
     2401 gccccatacg atataagttg taattctcat gtttgacagc ttatcatcgc atgagcggat
     2461 ccttatcgct atcgattcac acaaaaaacc aacacacaga tgtaatgaaa ataaagatat
     2521 tttattgcgg ccatcgtgat ggcaggttgg gcgtcgcttg gtcggtcatt tcgaacccca
     2581 gagtcccgct cagaagaact cgtcaagaag gcgatagaag gcgatgcgct gcgaatcggg
     2641 agcggcgata ccgtaaagca cgaggaagcg gtcagcccat tcgccgccaa gctcttcagc
     2701 aatatcacgg gtagccaacg ctatgtcctg atagcggtcc gccacaccca gccggccaca
     2761 gtcgatgaat ccagaaaagc ggccattttc caccatgata ttcggcaagc aggcatcgcc
     2821 atgggtcacg acgagatcct cgccgtcggg catgcgcgcc ttgagcctgg cgaacagttc
     2881 ggctggcgcg agcccctgat gctcttcgtc cagatcatcc tgatcgacaa gaccggcttc
     2941 catccgagta cgtgctcgct cgatgcgatg tttcgcttgg tggtcgaatg ggcaggtagc
     3001 cggatcaagc gtatgcagcc gccgcattgc atcagccatg atggatactt tctcggcagg
     3061 agcaaggtga gatgacagga gatcctgccc cggcacttcg cccaatagca gccagtccct
     3121 tcccgcttca gtgacaacgt cgagcacagc tgcgcaagga acgcccgtcg tggccagcca
     3181 cgatagccgc gctgcctcgt cctgcagttc attcagggca ccggacaggt cggtcttgac
     3241 aaaaagaacc gggcgcccct gcgctgacag ccggaacacg gcggcatcag agcagccgat
     3301 tgtctgttgt gcccagtcat agccgaatag cctctccacc caagcggccg gagaacctgc
     3361 gtgcaatcca tcttgttcaa tcatgatctg cctcctgggt ttagttcctc accttgtcgt
     3421 attatactat gccgatatac tatgccgatg attaattgtc aacagttaat taagcttttt
     3481 gcaaaagcct aggcctccaa aaaagcctcc tcactacttc tggaatagct cagaggccga
     3541 ggcggcctcg gcctctgcat aaataaaaaa aattagtcag ccatggggcg gagaatgggc
     3601 ggaactgggc ggagttaggg gcgggatggg cggagttagg ggcgggacta tggttgctga
     3661 ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca
     3721 cctggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg
     3781 ggactttcca caccctaact gacacacatt ccacagctgg ttctttccgc ctcagaaggt
     3841 acctaaccaa gttcctcttt cagaggttat ttcaggccat ggtgctgcgc agatccgcgt
     3901 atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag gaaattgtaa
     3961 gcgttaatat tttgttaaaa ttcgatttta ccacatttgt agaggtttta cttgcttaaa
     4021 aaaacctccc acatctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta
     4081 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     4141 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     4201 atcatgtctg ctcgaagcgg ccgccccaag gggttatgct agttattgct cagcggtggc
     4261 agcagccaac tcagcttcct ttcgggcttt gttagcagcc ggatcagctt gcatgcctgc
     4321 aggtcgactc tagaggatcc ccgggtaccg agcccgaatt cgtttaaact tgggtttttg
     4381 cttgaaatgg agttgtatca tagtcttgaa aatgaattcc cttagaagtt gtgatcttca
     4441 taatagcacc atttaaagca tcatcttcaa tgagtgttat caatccattg gcaatcaatg
     4501 gtgggtccaa aattccatag tatttaatca tatccttgat atgatcctta tattctatat
     4561 attgtcccat gttttcttct ttttcaattg attcaaggat ggctgtgtta acaaagcctg
     4621 gacaaatggc attcagtctc acaccactgt tcataagatt agcagccaac gctgctgagc
     4681 gtgtgaatcc aactatgcca tgctttgaag cacaataaac cggctgctgt gcaacgggca
     4741 tgagtcctgc taaagatgac atattgataa tgatgccgcc ttcacctcca ttttgcttac
     4801 tcatgtaatc caaaccaaga taggttccac tgataacaga aaccaaatta atttgcagag
     4861 ttttttccca gtttttctca ttattcactc cagcattatt gaccaaaatg tccagtcttc
     4921 caaagtggtc tacaactttt ctaaaagtgt ctctcagttg ttgctggtca gccacatcgc
     4981 actggatgaa cagagtcttc tgaggttcaa actgctcatc cagggcagct ttacactgta
     5041 cacctgcttc aagattccaa tccaccagcg ctaccttggc gcccttaagc agcagcgcct
     5101 ctgcaaaggc tctgcctatg ccctgagccg cgccggtcac cagcgccact ttgccgttca
     5161 cgtgcatggc gatcgcgccg ctcgagcccg ccagaatgcg ttcgcacagc cgccagccgg
     5221 tcactccgtt gatggttact cggaacagca gggagccgtc ggggttgatc aggcgctcgt
     5281 cgataatttt gttgccgttc cacagggtcc ctgttacagt gatctttttg ccgtcgaaca
     5341 cggcgatgcc ttcatacggc cgtccgaaat agtcgatcat gttcggcgta accccgtcga
     5401 ttaccagtgt gccatagtgc aggatcacct t
//
