LOCUS       sequence-474936-        9776 bp    DNA     circular SYN 02-JUL-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 9776)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 9776)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 2, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: WRN-HiBiT Fusion Vector
FEATURES             Location/Qualifiers
     source          1..9776
                     /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(6855..6873)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    6910..7031
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6921..6940)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6975..6994
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        7276..7633
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        7276..7467
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(7299..7319)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      7484..7619
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        7648..7695
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             7727..8521
                     /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(7781..7800)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     8391..8410
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    8585..8633
                     /note="synthetic polyadenylation signal"
     rep_origin      8790..9335
                     /direction=RIGHT
                     /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     9236..9255
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9452..9469
                     /label=L4440
                     /note="L4440 vector, forward primer"
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 agcactgatg aatcccctaa tgatttttat caaaatcatt aagttaaggt agatacacat
     1921 cttgtcatat gatcaaatgg tttcgccaaa aatcaataat cagacaacaa aatgtgcgaa
     1981 ctcgatattt tacacgactc tctttaccaa ttctgccccg aattacactt aaaacgactc
     2041 aacagcttaa cgttggcttg ccacgcctta cttgactgta aaactctcac tcttaccgaa
     2101 cttggccgta acctgccaac caaagcgaga acaaaacata acatcaaacg aatcgaccga
     2161 ttgttaggta atcgtcacct ccacaaagag cgactcgctg tataccgttg gcatgctagc
     2221 tttatctgtt cgggcaatac gatgcccatt gtacttgttg actggtctga tatccgtgag
     2281 caaaaacggc ttatggtatt gcgagcttca gtcgcactac acggtcgttc tgttactctt
     2341 tatgagaaag cgttcccgct ttcagagcaa tgttcaaaga aagctcatga ccaatttcta
     2401 gccgaccttg cgagcattct accgagtaac accacaccgc tcattgtcag tgatgctggc
     2461 tttaaagtgc catggtataa atccgttgag aagctgggtt ggtactggtt aagtcgagta
     2521 agaggaaaag tacaatatgc agacctagga gcggaaaact ggaaacctat cagcaactta
     2581 catgatatgt catctagtca ctcaaagact ttaggctata agaggctgac taaaagcaac
     2641 ccaatctcat gccaaattct attgtataaa tctcgctcta aaggccgaaa aaatcagcgc
     2701 tcgacacgga ctcattgtca ccacccgtca cctaaaatct actcagcgtc ggcaaaggag
     2761 ccatggattc tagcaactaa cttacctgtt gaaattcgaa cacccaaaca acttgttaat
     2821 atctattcga agcgaatgca gattgaagaa accttccgag acttgaaaag tcctgcctac
     2881 ggactaggcc tacgccatag ccgaacgagc agctcagagc gttttgatat catgctgcta
     2941 atcgccctga tgcttcaact aacatgttgg cttgcgggcg ttcatgctca gaaacaaggt
     3001 tgggacaagc acttccaggc taacacagtc agaaatcgaa acgtactctc aacagttcgc
     3061 ttaggcatgg aagttttgcg gcattctggc tacacaataa caagggaaga ctcactcgtg
     3121 gctgcaaccc tgcttactca aaatctattc acacatggtt acgttttggg gaaattatga
     3181 ggggatctct cagggctaag catcttcctc atgctttcag taaattggaa aacccacgga
     3241 gggtttctat cttactaaag gatatttcag aaaatctata ttcactgagg aggatgataa
     3301 ttgggtctac taacattgag actgaactga ggcccagcaa taatttaaac ttattatcct
     3361 ttgaagattc aactactggg ggagtacaac agaaacaaat tagagaacat gaagttttaa
     3421 ttcacgttga agatgaaaca tgggacccaa cacttgatca tttagctaaa catgatggag
     3481 aagatgtact tggaaataaa gtggaacgaa aagaagatgg atttgaagat ggagtagaag
     3541 acaacaaatt gaaagagaat atggaaagag cttgtttgat gtcgttagat attacagaac
     3601 atgaactcca aattttggaa cagcagtctc aggaagaata tcttagtgat attgcttata
     3661 aatctactga gcatttatct cccaatgata atgaaaacga tacgtcctat gtaattgaga
     3721 gtgatgaaga tttagaaatg gagatgctta agcatttatc tcccaatgat aatgaaaacg
     3781 atacgtccta tgtaattgag agtgatgaag atttagaaat ggagatgctt aagtctttag
     3841 aaaacctcaa tagtggcacg gtagaaccaa ctcattctaa atgcttaaaa atggaaagaa
     3901 atctgggtct tcctactaaa gaagaagaag aagatgatga aaatgaagct aatgaagggg
     3961 aagaagatga tgataaggac tttttgtggc cagcacccaa tgaagagcaa gttacttgcc
     4021 tcaagatgta ctttggccat tccagtttta aaccagttca gtggaaagtg attcattcag
     4081 tattagaaga aagaagagat aatgttgctg tcatggcaac tggatatgga aagagtttgt
     4141 gcttccagta tccacctgtt tatgtaggca agattggcct tgttatctct ccccttattt
     4201 ctctgatgga agaccaagtg ctacagctta aaatgtccaa catcccagct tgcttccttg
     4261 gatcagcaca gtcagaaaat gttctaacag atattaaatt aggtaaatac cggattgtat
     4321 acgtaactcc agaatactgt tcaggtaaca tgggcctgct ccagcaactt gaggctgata
     4381 ttggtatcac gctcattgct gtggatgagg ctcactgtat ttctgagtgg gggcatgatt
     4441 ttagggattc attcaggaag ttgggctccc taaagacagc actgccaatg gttccaatcg
     4501 ttgcacttac tgctactgca agttcttcaa tccgggaaga cattgtacgt tgcttaaatc
     4561 tgagaaatcc tcagatcacc tgtactggtt ttgatcgacc aaacctgtat ttagaagtta
     4621 ggcgaaaaac agggaatatc cttcaggatc tgcagccatt tcttgtcaaa acaagttccc
     4681 actgggaatt tgaaggtcca acaatcatct actgtccttc tagaaaaatg acacaacaag
     4741 ttacaggtga acttaggaaa ctgaatctat cctgtggaac ataccatgcg ggcatgagtt
     4801 ttagcacaag gaaagacatt catcataggt ttgtaagaga tgaaattcag tgtgtcatag
     4861 ctaccatagc ttttggaatg ggcattaata aagctgacat tcgccaagtc attcattacg
     4921 gtgctcctaa ggacatggaa tcatattatc aggagattgg tagagctggt cgtgatggac
     4981 ttcaaagttc ttgtcacgtc ctctgggctc ctgcagacat taacttaaat aggcaccttc
     5041 ttactgagat acgtaatgag aagtttcgat tatacaaatt aaagatgatg gcaaagatgg
     5101 aaaaatatct tcattctagc agatgtagga gacaaatcat cttgtctcat tttgaggaca
     5161 aacaagtaca aaaagcctcc ttgggaatta tgggaactga aaaatgctgt gataattgca
     5221 ggtccagatt ggatcattgc tattccatgg atgactcaga ggatacatcc tgggactttg
     5281 gtccacaagc atttaagctt ttgtctgctg tggacatctt aggcgaaaaa tttggaattg
     5341 ggcttccaat tttatttctc cgaggatcta attctcagcg tcttgccgat caatatcgca
     5401 ggcacagttt atttggcact ggcaaggatc aaacagagag ttggtggaag gctttttccc
     5461 gtcagctgat cactgaggga ttcttggtag aagtttctcg gtataacaaa tttatgaaga
     5521 tttgcgccct tacgaaaaag ggtagaaatt ggcttcataa agctaataca gaatctcaga
     5581 gcctcatcct tcaagctaat gaagaattgt gtccaaagaa gttgcttctg cctagttcga
     5641 aaactgtatc ttcgggcacc aaagagcatt gttataatca agtaccagtt gaattaagta
     5701 cagagaagaa gtctaacttg gagaagttat attcttataa accatgtgat aagatttctt
     5761 ctgggagtaa catttctaaa aaaagtatca tggtacagtc accagaaaaa gcttacagtt
     5821 cctcacagcc tgttatttcg gcacaagagc aggagactca gattgtgtta tatggcaaat
     5881 tggtagaagc taggcagaaa catgccaata aaatggatgt tcccccagct attctggcaa
     5941 caaacaagat actggtggat atggccaaaa tgagaccaac tacggttgaa aacgtaaaaa
     6001 ggattgatgg tgtttctgaa ggcaaagctg ccatgttggc ccctctgttg gaagtcatca
     6061 aacatttctg ccaaacaaat agtgttcaga cagacctctt ttcaagtaca aaacctcaag
     6121 aagaacagaa gacgagtctg gtagcaaaaa ataaaatatg cacactttca cagtctatgg
     6181 ccatcacata ctctttattc caagaaaaga agatgccttt gaagagcata gctgagagca
     6241 ggattctgcc tctcatgaca attggcatgc acttatccca agcggtgaaa gctggctgcc
     6301 cccttgattt ggagcgagca ggcctgactc cagaggttca gaagattatt gctgatgtta
     6361 tccgaaaccc tcccgtcaac tcagatatga gtaaaattag cctaatcaga atgttagttc
     6421 ctgaaaacat tgacacgtac cttatccaca tggcaattga gatccttaaa catggtcctg
     6481 acagcggact tcaaccttca tgtgatgtca acaaaaggag atgttttccc ggttctgaag
     6541 agatctgttc aagttctaag agaagcaagg aagaagtagg catcaatact gagacttcat
     6601 ctgcagagag aaagagacga ttacctgtgt ggtttgccaa aggaagtgat accagcaaga
     6661 aattaatgga caaaacgaaa aggggaggtc tttttagtgt ttctcaaggc agttcaggtg
     6721 gctcgagcgg tgtgagcggc tggcggctgt tcaagaagat tagctaaggc cgcgactcta
     6781 gagtcgacct gcaggcatgc aagctgatcc ggctgctaac aaagcccgaa aggaagctga
     6841 gttggctgct gccaccgctg agcaataact agcataaccc cttggggcgg ccgcttcgag
     6901 cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa
     6961 aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca
     7021 ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag ggggagatgt
     7081 gggaggtttt tttaagcaag taaaacctct acaaatgtgg taaaatcgaa ttttaacaaa
     7141 atattaacgc ttacaatttc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     7201 cacaccgcat acgcggatct gcgcagcacc atggcctgaa ataacctctg aaagaggaac
     7261 ttggttaggt accttctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg
     7321 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     7381 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     7441 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     7501 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     7561 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     7621 aggcttttgc aaaaagctta attaactgtt gacaattaat catcggcata gtatatcggc
     7681 atagtataat acgacaaggt gaggaactaa acccaggagg cagatcatga ttgaacaaga
     7741 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     7801 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     7861 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc
     7921 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     7981 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     8041 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     8101 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     8161 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     8221 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt
     8281 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     8341 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     8401 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     8461 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     8521 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgatgg
     8581 ccgcaataaa atatctttat tttcattaca tctgtgtgtt ggttttttgt gtgaatcgat
     8641 agcgataagg atccgctcat gcgatgataa gctgtcaaac atgagaatta caacttatat
     8701 cgtatggggc tgacttcagg tgctacattt gaagagataa attgcactga aatctagaaa
     8761 tattttatct gattaataag atgatcttct tgagatcgtt ttggtctgcg cgtaatctct
     8821 tgctctgaaa acgaaaaaac cgccttgcag ggcggttttt cgaaggttct ctgagctacc
     8881 aactctttga accgaggtaa ctggcttgga ggagcgcagt caccaaaact tgtcctttca
     8941 gtttagcctt aaccggcgca tgacttcaag actaactcct ctaaatcaat taccagtggc
     9001 tgctgccagt ggtgcttttg catgtctttc cgggttggac tcaagacgat agttaccgga
     9061 taaggcgcag cggtcggact gaacgggggg ttcgtgcata cagtccagct tggagcgaac
     9121 tgcctacccg gaactgagtg tcaggcgtgg aatgagacaa acgcggccat aacagcggaa
     9181 tgacaccggt aaaccgaaag gcaggaacag gagagcgcac gagggagccg ccagggggaa
     9241 acgcctggta tctttatagt cctgtcgggt ttcgccacca ctgatttgag cgtcagattt
     9301 cgtgatgctt gtcagggggg cggagcctat ggaaaaacgg ctttgccgcg gccctctcac
     9361 ttccctgtta agtatcttcc tggcatcttc caggaaatct ccgccccgtt cgtaagccat
     9421 ttccgctcgc cgcagtcgaa cgaccgagcg tagcgagtca gtgagcgagg aagcggaata
     9481 tatcctgtat cacatattct gctgacgcac cggtgcagcc ttttttctcc tgccacatga
     9541 agcacttcac tgacaccctc atcagtgcca acatagtaag ccagtataca ctccgctagc
     9601 gctgatgtcc ggcggtgctt ttgccgttac gcaccacccc gtcatgagat tacgcgtttt
     9661 agcggtgcgt acaattaagg gattatggta aatccactta ctgtctgccc tcgtagccat
     9721 cgagataaac cgcagtactc cggccacgat gcgtccggcg tagaggatcg agatct
//
