LOCUS       sequence-470996-        8574 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express WRNu-HiBiT in Mammalian Cells under a CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8574)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8574)
  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            "Unpublished"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: WRN-HiBiT Fusion Vector
FEATURES             Location/Qualifiers
     source          1..8574
                     /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"
     primer_bind     complement(5517..5535)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    5572..5693
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5583..5602)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5637..5656
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        5938..6295
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5938..6129
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5961..5981)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6146..6281
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        6310..6357
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             6389..7183
                     /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(6443..6462)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7053..7072
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7247..7295
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(7534..8122)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7614..7633)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    8238..8521
                     /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 ggctagcgat cgccatgagt gaaaaaaaat
     1081 tggaaacaac tgcacagcag cggaaatgtc ctgaatggat gaatgtgcag aataaaagat
     1141 gtgctgtaga agaaagaaag gcatgtgttc ggaagagtgt ttttgaagat gacctcccct
     1201 tcttagaatt cactggatcc attgtgtata gttacgatgc tagtgattgc tctttcctgt
     1261 cagaagatat tagcatgagt ctatcagatg gggatgtggt gggatttgac atggagtggc
     1321 caccattata caatagaggg aaacttggca aagttgcact aattcagttg tgtgtttctg
     1381 agagcaaatg ttacttgttc cacgtttctt ccatgtcagt ttttccccag ggattaaaaa
     1441 tgttgcttga aaataaagca gttaaaaagg caggtgtagg aattgaagga gatcagtgga
     1501 aacttctacg tgactttgat atcaaattga agaattttgt ggagttgaca gatgttgcca
     1561 ataaaaagct gaaatgcaca gagacctgga gccttaacag tctggttaaa cacctcttag
     1621 gtaaacagct cctgaaagac aagtctatcc gctgtagcaa ttggagtaaa tttcctctca
     1681 ctgaggacca gaaactgtat gcagccactg atgcttatgc tggttttatt atttaccgaa
     1741 atttagagat tttggatgat actgtgcaaa ggtttgctat aaataaagag gaagaaatcc
     1801 tacttagcga catgaacaaa cagttgactt caatctctga ggaagtgatg gatctggcta
     1861 agcatcttcc tcatgctttc agtaaattgg aaaacccacg gagggtttct atcttactaa
     1921 aggatatttc agaaaatcta tattcactga ggaggatgat aattgggtct actaacattg
     1981 agactgaact gaggcccagc aataatttaa acttattatc ctttgaagat tcaactactg
     2041 ggggagtaca acagaaacaa attagagaac atgaagtttt aattcacgtt gaagatgaaa
     2101 catgggaccc aacacttgat catttagcta aacatgatgg agaagatgta cttggaaata
     2161 aagtggaacg aaaagaagat ggatttgaag atggagtaga agacaacaaa ttgaaagaga
     2221 atatggaaag agcttgtttg atgtcgttag atattacaga acatgaactc caaattttgg
     2281 aacagcagtc tcaggaagaa tatcttagtg atattgctta taaatctact gagcatttat
     2341 ctcccaatga taatgaaaac gatacgtcct atgtaattga gagtgatgaa gatttagaaa
     2401 tggagatgct taagcattta tctcccaatg ataatgaaaa cgatacgtcc tatgtaattg
     2461 agagtgatga agatttagaa atggagatgc ttaagtcttt agaaaacctc aatagtggca
     2521 cggtagaacc aactcattct aaatgcttaa aaatggaaag aaatctgggt cttcctacta
     2581 aagaagaaga agaagatgat gaaaatgaag ctaatgaagg ggaagaagat gatgataagg
     2641 actttttgtg gccagcaccc aatgaagagc aagttacttg cctcaagatg tactttggcc
     2701 attccagttt taaaccagtt cagtggaaag tgattcattc agtattagaa gaaagaagag
     2761 ataatgttgc tgtcatggca actggatatg gaaagagttt gtgcttccag tatccacctg
     2821 tttatgtagg caagattggc cttgttatct ctccccttat ttctctgatg gaagaccaag
     2881 tgctacagct taaaatgtcc aacatcccag cttgcttcct tggatcagca cagtcagaaa
     2941 atgttctaac agatattaaa ttaggtaaat accggattgt atacgtaact ccagaatact
     3001 gttcaggtaa catgggcctg ctccagcaac ttgaggctga tattggtatc acgctcattg
     3061 ctgtggatga ggctcactgt atttctgagt gggggcatga ttttagggat tcattcagga
     3121 agttgggctc cctaaagaca gcactgccaa tggttccaat cgttgcactt actgctactg
     3181 caagttcttc aatccgggaa gacattgtac gttgcttaaa tctgagaaat cctcagatca
     3241 cctgtactgg ttttgatcga ccaaacctgt atttagaagt taggcgaaaa acagggaata
     3301 tccttcagga tctgcagcca tttcttgtca aaacaagttc ccactgggaa tttgaaggtc
     3361 caacaatcat ctactgtcct tctagaaaaa tgacacaaca agttacaggt gaacttagga
     3421 aactgaatct atcctgtgga acataccatg cgggcatgag ttttagcaca aggaaagaca
     3481 ttcatcatag gtttgtaaga gatgaaattc agtgtgtcat agctaccata gcttttggaa
     3541 tgggcattaa taaagctgac attcgccaag tcattcatta cggtgctcct aaggacatgg
     3601 aatcatatta tcaggagatt ggtagagctg gtcgtgatgg acttcaaagt tcttgtcacg
     3661 tcctctgggc tcctgcagac attaacttaa ataggcacct tcttactgag atacgtaatg
     3721 agaagtttcg attatacaaa ttaaagatga tggcaaagat ggaaaaatat cttcattcta
     3781 gcagatgtag gagacaaatc atcttgtctc attttgagga caaacaagta caaaaagcct
     3841 ccttgggaat tatgggaact gaaaaatgct gtgataattg caggtccaga ttggatcatt
     3901 gctattccat ggatgactca gaggatacat cctgggactt tggtccacaa gcatttaagc
     3961 ttttgtctgc tgtggacatc ttaggcgaaa aatttggaat tgggcttcca attttatttc
     4021 tccgaggatc taattctcag cgtcttgccg atcaatatcg caggcacagt ttatttggca
     4081 ctggcaagga tcaaacagag agttggtgga aggctttttc ccgtcagctg atcactgagg
     4141 gattcttggt agaagtttct cggtataaca aatttatgaa gatttgcgcc cttacgaaaa
     4201 agggtagaaa ttggcttcat aaagctaata cagaatctca gagcctcatc cttcaagcta
     4261 atgaagaatt gtgtccaaag aagttgcttc tgcctagttc gaaaactgta tcttcgggca
     4321 ccaaagagca ttgttataat caagtaccag ttgaattaag tacagagaag aagtctaact
     4381 tggagaagtt atattcttat aaaccatgtg ataagatttc ttctgggagt aacatttcta
     4441 aaaaaagtat catggtacag tcaccagaaa aagcttacag ttcctcacag cctgttattt
     4501 cggcacaaga gcaggagact cagattgtgt tatatggcaa attggtagaa gctaggcaga
     4561 aacatgccaa taaaatggat gttcccccag ctattctggc aacaaacaag atactggtgg
     4621 atatggccaa aatgagacca actacggttg aaaacgtaaa aaggattgat ggtgtttctg
     4681 aaggcaaagc tgccatgttg gcccctctgt tggaagtcat caaacatttc tgccaaacaa
     4741 atagtgttca gacagacctc ttttcaagta caaaacctca agaagaacag aagacgagtc
     4801 tggtagcaaa aaataaaata tgcacacttt cacagtctat ggccatcaca tactctttat
     4861 tccaagaaaa gaagatgcct ttgaagagca tagctgagag caggattctg cctctcatga
     4921 caattggcat gcacttatcc caagcggtga aagctggctg cccccttgat ttggagcgag
     4981 caggcctgac tccagaggtt cagaagatta ttgctgatgt tatccgaaac cctcccgtca
     5041 actcagatat gagtaaaatt agcctaatca gaatgttagt tcctgaaaac attgacacgt
     5101 accttatcca catggcaatt gagatcctta aacatggtcc tgacagcgga cttcaacctt
     5161 catgtgatgt caacaaaagg agatgttttc ccggttctga agagatctgt tcaagttcta
     5221 agagaagcaa ggaagaagta ggcatcaata ctgagacttc atctgcagag agaaagagac
     5281 gattacctgt gtggtttgcc aaaggaagtg ataccagcaa gaaattaatg gacaaaacga
     5341 aaaggggagg tctttttagt gtttctcaag gcagttcagg tggctcgagc ggtgtgagcg
     5401 gctggcggct gttcaagaag attagctaag gccgcgactc tagagtcgac ctgcaggcat
     5461 gcaagctgat ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc
     5521 tgagcaataa ctagcataac cccttggggc ggccgcttcg agcagacatg ataagataca
     5581 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
     5641 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
     5701 acaattgcat tcattttatg tttcaggttc agggggagat gtgggaggtt tttttaagca
     5761 agtaaaacct ctacaaatgt ggtaaaatcg aattttaaca aaatattaac gcttacaatt
     5821 tcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgcggat
     5881 ctgcgcagca ccatggcctg aaataacctc tgaaagagga acttggttag gtaccttctg
     5941 aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     6001 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     6061 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     6121 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     6181 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     6241 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     6301 taattaactg ttgacaatta atcatcggca tagtatatcg gcatagtata atacgacaag
     6361 gtgaggaact aaacccagga ggcagatcat gattgaacaa gatggattgc acgcaggttc
     6421 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     6481 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     6541 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc
     6601 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     6661 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     6721 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     6781 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     6841 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     6901 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     6961 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     7021 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     7081 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     7141 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     7201 gaaatgaccg accaagcgac gcccaacctg ccatcacgat ggccgcaata aaatatcttt
     7261 attttcatta catctgtgtg ttggtttttt gtgtgaatcg atagcgataa ggatcctctt
     7321 tgcgcttgcg ttttcccttg tccagatagc ccagtagctg acattcatcc ggggtcagca
     7381 ccgtttctgc ggactggctt tctacccggt atcagctcac tcaaaggcgg taatacggtt
     7441 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     7501 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     7561 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     7621 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     7681 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     7741 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     7801 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     7861 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     7921 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     7981 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     8041 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     8101 gcgcagaaaa aaaggatttc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     8161 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     8221 ctagatcctt ttatagtccg gaaatacagg aacgcacgct ggatggccct tcgctgggat
     8281 ggtgaaacca tgaaaaatgg cagcttcagt ggattaagtg ggggtaatgt ggcctgtacc
     8341 ctctggttgc ataggtattc atacggttaa aatttatcag gcgcgattgc ggcagttttt
     8401 cgggtggttt gttgccattt ttacctgtct gctgccgtga tcgcgctgaa cgcgttttag
     8461 cggtgcgtac aattaaggga ttatggtaaa tccacttact gtctgccctc gtagccatcg
     8521 agataaaccg cagtactccg gccacgatgc gtccggcgta gaggatcgag atct
//
