LOCUS       sequence-474980-        5889 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express NanoLuc(R)-UHMK1 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 5889)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5889)
  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            "Unpublished"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-UHMK1 Fusion Vector
FEATURES             Location/Qualifiers
     source          1..5889
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(899..1411)
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     primer_bind     complement(1424..1443)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1425..1443)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1485..1504)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     intron          1487..1619
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        complement(1755..1958)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1784..1804)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1959..2338
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(2783..2800)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2917..3462)
                     /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(2997..3016)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3619..3667
                     /note="synthetic polyadenylation signal"
     CDS             complement(3731..4525)
                     /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(3842..3861)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4452..4471
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4557..4604)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(4619..4976)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4633..4768
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        4785..4976
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     4933..4953
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    5221..5342
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5258..5277)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5312..5331
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5379..5397
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
ORIGIN
        1 ttctccggct tggatcgtcg tgcagcatag acttgatcag gtccctcagg tggtatgcag
       61 ggattgctgc attcaccacg gccttgctgg caaagatgtg atcgatgatg gcagaggagt
      121 tggccttcca ctcctgagag cgcactgtgt gcttcagctt catgccggag aacatctcca
      181 gcaggatgat gcccagggac cacaggtcca cggcgcttgt acactcggta tcagactgca
      241 ggccggcctg ggccaggcaa ttctgcagct cggcctcagg ggctctatag ccgtcggtct
      301 ggatgtactt cacatcctgg ttgccctcct taaagctcag gccgaagtcg atcagcttaa
      361 aacactcatt ctcggcagac cacaggatgt tcctgggctt caggtcggcg tgcacatagc
      421 cctcgtggtg caggaaggcc agggcctcca gcacatctct ggcacagtgc tggatcatcc
      481 acatagagca gccctggtgg ctagagtaca gcagcagctc ggacacgctc acatccagca
      541 gctccagcag caggcacctg gatggcacat ttgggctaaa gtggattgtg aacacgccat
      601 acagggtcac gatgttcctg tgtccctgca gctgctccag ggcggctctc tccttccgaa
      661 agccgtactc tgcggcggat gctgctgctc ctgtggttcc tggaggcagg aactgcttca
      721 gggcgcctgg tggggatcca gggtttccac agcagcgcac cctgtacacg ctggcggagc
      781 tgccgcttcc cagcctagac tgcacctgcc acaggcggcc aaaggcctcc aggaacctag
      841 ggggctctgc tccccatgcg catccgcttc ctgccatggc gatcgcgccg ctcgagcccg
      901 ccagaatgcg ttcgcacagc cgccagccgg tcactccgtt gatggttact cggaacagca
      961 gggagccgtc ggggttgatc aggcgctcgt cgataatttt gttgccgttc cacagggtcc
     1021 ctgttacagt gatctttttg ccgtcgaaca cggcgatgcc ttcatacggc cgtccgaaat
     1081 agtcgatcat gttcggcgta accccgtcga ttaccagtgt gccatagtgc aggatcacct
     1141 taaagtgatg atcatccaca gggtacacca ccttaaaaat tttttcgatc tggcccattt
     1201 ggtcgccgct cagaccttca tacgggatga tgacatggat gtcgatcttc agcccatttt
     1261 caccgctcag gacaatcctt tggatcggag ttacggacac cccgagattc tgaaacaaac
     1321 tggacacacc tccctgttca aggacttggt ccaggttgta gccggctgtc tgtcgccagt
     1381 ccccaacgaa atcttcgagt gtgaagacca tggtgagcgc tagccctata gtgagtcgta
     1441 ttaatactct agccttaaga gctgtaattg aactgggagt ggacacctgt ggagagaaag
     1501 gcaaagtgga tgtcagtaag accaataggt gcctatcaga aacgcaagag tcttctctgt
     1561 ctcgacaagc ccagtttcta ttggtctcct taaacctgtc ttgtaacctt gatacttacc
     1621 tgcccagtgc ctcacgacca acttctgcag cttaagttcg agactgttgt gtcagaagca
     1681 ctgactgcgt tagcaattta actgtgataa actaccgcaa taaagcttct agtgatctga
     1741 cggttcacta aaccagctct gcttatatag acctcccacc gtacacgcct accgcccatt
     1801 tgcgtcaacg gggcggggtt attacgacat tttggaaagt cccgttgatt ttggtgccaa
     1861 aacaaactcc cattgacgtc aatggggtgg agacttggaa atccccgtga gtcaaaccgc
     1921 tatccacgcc cattggtgta ctgccaaaac cgcatcacca tggtaatagc gatgactaat
     1981 acgtagatgt actgccaagt aggaaagtcc cgtaaggtca tgtactgggc ataatgccag
     2041 gcgggccatt taccgtcatt gacgtcaata gggggcggac ttggcatatg atacacttga
     2101 tgtactgcca agtgggcagt ttaccgtaaa tactccaccc attgacgtca atggaaagtc
     2161 cctattggcg ttactatggg aacatacgtc attattgacg tcaatgggcg ggggtcgttg
     2221 ggcggtcagc caggcgggcc atttaccgta agttatgtaa cgcggaactc catatatggg
     2281 ctatgaacta atgaccccgt aattgattac tattaataac tagtcaataa tcaatgccaa
     2341 catggcggtc atattggaca tgagccaata taaatgtaca tattatgata tagatacaac
     2401 gtatgcaatg gccaatagcc aatattgatt tatgctatat aaccaatgaa taatatggct
     2461 aatggccaat attgaagatc tcgatcctct acgccggacg catcgtggcc ggagtactgc
     2521 ggtttatctc gatggctacg agggcagaca gtaagtggat ttaccataat cccttaattg
     2581 tacgcaccgc taaaacgcgt aatctcatga cggggtggtg cgtaacggca aaagcaccgc
     2641 cggacatcag cgctagcgga gtgtatactg gcttactatg ttggcactga tgagggtgtc
     2701 agtgaagtgc ttcatgtggc aggagaaaaa aggctgcacc ggtgcgtcag cagaatatgt
     2761 gatacaggat atattccgct tcctcgctca ctgactcgct acgctcggtc gttcgactgc
     2821 ggcgagcgga aatggcttac gaacggggcg gagatttcct ggaagatgcc aggaagatac
     2881 ttaacaggga agtgagaggg ccgcggcaaa gccgtttttc cataggctcc gcccccctga
     2941 caagcatcac gaaatctgac gctcaaatca gtggtggcga aacccgacag gactataaag
     3001 ataccaggcg tttccccctg gcggctccct cgtgcgctct cctgttcctg cctttcggtt
     3061 taccggtgtc attccgctgt tatggccgcg tttgtctcat tccacgcctg acactcagtt
     3121 ccgggtaggc agttcgctcc aagctggact gtatgcacga accccccgtt cagtccgacc
     3181 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggaaagacat gcaaaagcac
     3241 cactggcagc agccactggt aattgattta gaggagttag tcttgaagtc atgcgccggt
     3301 taaggctaaa ctgaaaggac aagttttggt gactgcgctc ctccaagcca gttacctcgg
     3361 ttcaaagagt tggtagctca gagaaccttc gaaaaaccgc cctgcaaggc ggttttttcg
     3421 ttttcagagc aagagattac gcgcagacca aaacgatctc aagaagatca tcttattaat
     3481 cagataaaat atttctagat ttcagtgcaa tttatctctt caaatgtagc acctgaagtc
     3541 agccccatac gatataagtt gtaattctca tgtttgacag cttatcatcg catgagcgga
     3601 tccttatcgc tatcgattca cacaaaaaac caacacacag atgtaatgaa aataaagata
     3661 ttttattgcg gccatcgtga tggcaggttg ggcgtcgctt ggtcggtcat ttcgaacccc
     3721 agagtcccgc tcagaagaac tcgtcaagaa ggcgatagaa ggcgatgcgc tgcgaatcgg
     3781 gagcggcgat accgtaaagc acgaggaagc ggtcagccca ttcgccgcca agctcttcag
     3841 caatatcacg ggtagccaac gctatgtcct gatagcggtc cgccacaccc agccggccac
     3901 agtcgatgaa tccagaaaag cggccatttt ccaccatgat attcggcaag caggcatcgc
     3961 catgggtcac gacgagatcc tcgccgtcgg gcatgcgcgc cttgagcctg gcgaacagtt
     4021 cggctggcgc gagcccctga tgctcttcgt ccagatcatc ctgatcgaca agaccggctt
     4081 ccatccgagt acgtgctcgc tcgatgcgat gtttcgcttg gtggtcgaat gggcaggtag
     4141 ccggatcaag cgtatgcagc cgccgcattg catcagccat gatggatact ttctcggcag
     4201 gagcaaggtg agatgacagg agatcctgcc ccggcacttc gcccaatagc agccagtccc
     4261 ttcccgcttc agtgacaacg tcgagcacag ctgcgcaagg aacgcccgtc gtggccagcc
     4321 acgatagccg cgctgcctcg tcctgcagtt cattcagggc accggacagg tcggtcttga
     4381 caaaaagaac cgggcgcccc tgcgctgaca gccggaacac ggcggcatca gagcagccga
     4441 ttgtctgttg tgcccagtca tagccgaata gcctctccac ccaagcggcc ggagaacctg
     4501 cgtgcaatcc atcttgttca atcatgatct gcctcctggg tttagttcct caccttgtcg
     4561 tattatacta tgccgatata ctatgccgat gattaattgt caacagttaa ttaagctttt
     4621 tgcaaaagcc taggcctcca aaaaagcctc ctcactactt ctggaatagc tcagaggccg
     4681 aggcggcctc ggcctctgca taaataaaaa aaattagtca gccatggggc ggagaatggg
     4741 cggaactggg cggagttagg ggcgggatgg gcggagttag gggcgggact atggttgctg
     4801 actaattgag atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac
     4861 acctggttgc tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg
     4921 gggactttcc acaccctaac tgacacacat tccacagctg gttctttccg cctcagaagg
     4981 tacctaacca agttcctctt tcagaggtta tttcaggcca tggtgctgcg cagatccgcg
     5041 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggaaattgta
     5101 agcgttaata ttttgttaaa attcgatttt accacatttg tagaggtttt acttgcttaa
     5161 aaaaacctcc cacatctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt
     5221 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     5281 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     5341 tatcatgtct gctcgaagcg gccgccccaa ggggttatgc tagttattgc tcagcggtgg
     5401 cagcagccaa ctcagcttcc tttcgggctt tgttagcagc cggatcagct tgcatgcctg
     5461 caggtcgact ctagaggatc cccgggtacc gagcccgaat tcgtttaaac cagcagtgtc
     5521 tgatacaggt agcccctctt ataggcggac agggggtaga aggtggccac cacaaacttg
     5581 ccgtcgaaca ttctgccggt cagcagcttc tgggcggcct tagaatcgcc ggcattggca
     5641 tactccacga acacctggcc cctgccaggg ttctccttgg gcaccagcag ggacaccact
     5701 gggccgtact tctgacactc ctccttcacg tcctccacca catcctcata ctcctcctca
     5761 ttctccaggt agtcatcgtc cagcacgttc agcagtctca gcacaggggt gggcagcatg
     5821 accaggtcct cgatgtgtgg ggcgaaaggg atagaaaaga atggggagca cagggccatc
     5881 tcggcgggg
//
