LOCUS       sequence-474940-        8464 bp    DNA     circular SYN 02-JUL-2025
DEFINITION  Express HiBiT-WRN in Mammalian Cells under a CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8464)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 8464)
  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: HiBiT-WRN Fusion Vector
FEATURES             Location/Qualifiers
     source          1..8464
                     /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(5543..5561)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    5598..5719
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5609..5628)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5663..5682
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        5964..6321
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5964..6155
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5987..6007)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6172..6307
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        6336..6383
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             6415..7209
                     /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(6469..6488)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7079..7098
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7273..7321
                     /note="synthetic polyadenylation signal"
     rep_origin      7478..8023
                     /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     7924..7943
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8140..8157
                     /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 ggctagcgct caccatggtg agcggctggc
     1081 ggctgttcaa gaagattagc gggagctccg gtggctcgag cggtgcgatc gccatgagtg
     1141 aaaaaaaatt ggaaacaact gcacagcagc ggaaatgtcc tgaatggatg aatgtgcaga
     1201 ataaaagatg tgctgtagaa gaaagaaagg catgtgttcg gaagagtgtt tttgaagatg
     1261 acctcccctt cttagaattc actggatcca ttgtgtatag ttacgatgct agtgattgct
     1321 ctttcctgtc agaagatatt agcatgagtc tatcagatgg ggatgtggtg ggatttgaca
     1381 tggagtggcc accattatac aatagaggga aacttggcaa agttgcacta attcagttgt
     1441 gtgtttctga gagcaaatgt tacttgttcc acgtttcttc catgtcagtt tttccccagg
     1501 gattaaaaat gttgcttgaa aataaagcag ttaaaaaggc aggtgtagga attgaaggag
     1561 atcagtggaa acttctacgt gactttgata tcaaattgaa gaattttgtg gagttgacag
     1621 atgttgccaa taaaaagctg aaatgcacag agacctggag ccttaacagt ctggttaaac
     1681 acctcttagg taaacagctc ctgaaagaca agtctatccg ctgtagcaat tggagtaaat
     1741 ttcctctcac tgaggaccag aaactgtatg cagccactga tgcttatgct ggttttatta
     1801 tttaccgaaa tttagagatt ttggatgata ctgtgcaaag gtttgctata aataaagagg
     1861 aagaaatcct acttagcgac atgaacaaac agttgacttc aatctctgag gaagtgatgg
     1921 atctggctaa gcatcttcct catgctttca gtaaattgga aaacccacgg agggtttcta
     1981 tcttactaaa ggatatttca gaaaatctat attcactgag gaggatgata attgggtcta
     2041 ctaacattga gactgaactg aggcccagca ataatttaaa cttattatcc tttgaagatt
     2101 caactactgg gggagtacaa cagaaacaaa ttagagaaca tgaagtttta attcacgttg
     2161 aagatgaaac atgggaccca acacttgatc atttagctaa acatgatgga gaagatgtac
     2221 ttggaaataa agtggaacga aaagaagatg gatttgaaga tggagtagaa gacaacaaat
     2281 tgaaagagaa tatggaaaga gcttgtttga tgtcgttaga tattacagaa catgaactcc
     2341 aaattttgga acagcagtct caggaagaat atcttagtga tattgcttat aaatctactg
     2401 agcatttatc tcccaatgat aatgaaaacg atacgtccta tgtaattgag agtgatgaag
     2461 atttagaaat ggagatgctt aagcatttat ctcccaatga taatgaaaac gatacgtcct
     2521 atgtaattga gagtgatgaa gatttagaaa tggagatgct taagtcttta gaaaacctca
     2581 atagtggcac ggtagaacca actcattcta aatgcttaaa aatggaaaga aatctgggtc
     2641 ttcctactaa agaagaagaa gaagatgatg aaaatgaagc taatgaaggg gaagaagatg
     2701 atgataagga ctttttgtgg ccagcaccca atgaagagca agttacttgc ctcaagatgt
     2761 actttggcca ttccagtttt aaaccagttc agtggaaagt gattcattca gtattagaag
     2821 aaagaagaga taatgttgct gtcatggcaa ctggatatgg aaagagtttg tgcttccagt
     2881 atccacctgt ttatgtaggc aagattggcc ttgttatctc tccccttatt tctctgatgg
     2941 aagaccaagt gctacagctt aaaatgtcca acatcccagc ttgcttcctt ggatcagcac
     3001 agtcagaaaa tgttctaaca gatattaaat taggtaaata ccggattgta tacgtaactc
     3061 cagaatactg ttcaggtaac atgggcctgc tccagcaact tgaggctgat attggtatca
     3121 cgctcattgc tgtggatgag gctcactgta tttctgagtg ggggcatgat tttagggatt
     3181 cattcaggaa gttgggctcc ctaaagacag cactgccaat ggttccaatc gttgcactta
     3241 ctgctactgc aagttcttca atccgggaag acattgtacg ttgcttaaat ctgagaaatc
     3301 ctcagatcac ctgtactggt tttgatcgac caaacctgta tttagaagtt aggcgaaaaa
     3361 cagggaatat ccttcaggat ctgcagccat ttcttgtcaa aacaagttcc cactgggaat
     3421 ttgaaggtcc aacaatcatc tactgtcctt ctagaaaaat gacacaacaa gttacaggtg
     3481 aacttaggaa actgaatcta tcctgtggaa cataccatgc gggcatgagt tttagcacaa
     3541 ggaaagacat tcatcatagg tttgtaagag atgaaattca gtgtgtcata gctaccatag
     3601 cttttggaat gggcattaat aaagctgaca ttcgccaagt cattcattac ggtgctccta
     3661 aggacatgga atcatattat caggagattg gtagagctgg tcgtgatgga cttcaaagtt
     3721 cttgtcacgt cctctgggct cctgcagaca ttaacttaaa taggcacctt cttactgaga
     3781 tacgtaatga gaagtttcga ttatacaaat taaagatgat ggcaaagatg gaaaaatatc
     3841 ttcattctag cagatgtagg agacaaatca tcttgtctca ttttgaggac aaacaagtac
     3901 aaaaagcctc cttgggaatt atgggaactg aaaaatgctg tgataattgc aggtccagat
     3961 tggatcattg ctattccatg gatgactcag aggatacatc ctgggacttt ggtccacaag
     4021 catttaagct tttgtctgct gtggacatct taggcgaaaa atttggaatt gggcttccaa
     4081 ttttatttct ccgaggatct aattctcagc gtcttgccga tcaatatcgc aggcacagtt
     4141 tatttggcac tggcaaggat caaacagaga gttggtggaa ggctttttcc cgtcagctga
     4201 tcactgaggg attcttggta gaagtttctc ggtataacaa atttatgaag atttgcgccc
     4261 ttacgaaaaa gggtagaaat tggcttcata aagctaatac agaatctcag agcctcatcc
     4321 ttcaagctaa tgaagaattg tgtccaaaga agttgcttct gcctagttcg aaaactgtat
     4381 cttcgggcac caaagagcat tgttataatc aagtaccagt tgaattaagt acagagaaga
     4441 agtctaactt ggagaagtta tattcttata aaccatgtga taagatttct tctgggagta
     4501 acatttctaa aaaaagtatc atggtacagt caccagaaaa agcttacagt tcctcacagc
     4561 ctgttatttc ggcacaagag caggagactc agattgtgtt atatggcaaa ttggtagaag
     4621 ctaggcagaa acatgccaat aaaatggatg ttcccccagc tattctggca acaaacaaga
     4681 tactggtgga tatggccaaa atgagaccaa ctacggttga aaacgtaaaa aggattgatg
     4741 gtgtttctga aggcaaagct gccatgttgg cccctctgtt ggaagtcatc aaacatttct
     4801 gccaaacaaa tagtgttcag acagacctct tttcaagtac aaaacctcaa gaagaacaga
     4861 agacgagtct ggtagcaaaa aataaaatat gcacactttc acagtctatg gccatcacat
     4921 actctttatt ccaagaaaag aagatgcctt tgaagagcat agctgagagc aggattctgc
     4981 ctctcatgac aattggcatg cacttatccc aagcggtgaa agctggctgc ccccttgatt
     5041 tggagcgagc aggcctgact ccagaggttc agaagattat tgctgatgtt atccgaaacc
     5101 ctcccgtcaa ctcagatatg agtaaaatta gcctaatcag aatgttagtt cctgaaaaca
     5161 ttgacacgta ccttatccac atggcaattg agatccttaa acatggtcct gacagcggac
     5221 ttcaaccttc atgtgatgtc aacaaaagga gatgttttcc cggttctgaa gagatctgtt
     5281 caagttctaa gagaagcaag gaagaagtag gcatcaatac tgagacttca tctgcagaga
     5341 gaaagagacg attacctgtg tggtttgcca aaggaagtga taccagcaag aaattaatgg
     5401 acaaaacgaa aaggggaggt ctttttagtg tttaaacgaa ttcgggctcg gtacccgggg
     5461 atcctctaga gtcgacctgc aggcatgcaa gctgatccgg ctgctaacaa agcccgaaag
     5521 gaagctgagt tggctgctgc caccgctgag caataactag cataacccct tggggcggcc
     5581 gcttcgagca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca
     5641 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
     5701 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg
     5761 ggagatgtgg gaggtttttt taagcaagta aaacctctac aaatgtggta aaatcgaatt
     5821 ttaacaaaat attaacgctt acaatttcct gatgcggtat tttctcctta cgcatctgtg
     5881 cggtatttca caccgcatac gcggatctgc gcagcaccat ggcctgaaat aacctctgaa
     5941 agaggaactt ggttaggtac cttctgaggc ggaaagaacc agctgtggaa tgtgtgtcag
     6001 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     6061 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     6121 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     6181 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     6241 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     6301 ggaggcctag gcttttgcaa aaagcttaat taactgttga caattaatca tcggcatagt
     6361 atatcggcat agtataatac gacaaggtga ggaactaaac ccaggaggca gatcatgatt
     6421 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     6481 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     6541 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     6601 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     6661 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     6721 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     6781 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     6841 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     6901 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     6961 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     7021 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     7081 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     7141 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     7201 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     7261 cacgatggcc gcaataaaat atctttattt tcattacatc tgtgtgttgg ttttttgtgt
     7321 gaatcgatag cgataaggat ccgctcatgc gatgataagc tgtcaaacat gagaattaca
     7381 acttatatcg tatggggctg acttcaggtg ctacatttga agagataaat tgcactgaaa
     7441 tctagaaata ttttatctga ttaataagat gatcttcttg agatcgtttt ggtctgcgcg
     7501 taatctcttg ctctgaaaac gaaaaaaccg ccttgcaggg cggtttttcg aaggttctct
     7561 gagctaccaa ctctttgaac cgaggtaact ggcttggagg agcgcagtca ccaaaacttg
     7621 tcctttcagt ttagccttaa ccggcgcatg acttcaagac taactcctct aaatcaatta
     7681 ccagtggctg ctgccagtgg tgcttttgca tgtctttccg ggttggactc aagacgatag
     7741 ttaccggata aggcgcagcg gtcggactga acggggggtt cgtgcataca gtccagcttg
     7801 gagcgaactg cctacccgga actgagtgtc aggcgtggaa tgagacaaac gcggccataa
     7861 cagcggaatg acaccggtaa accgaaaggc aggaacagga gagcgcacga gggagccgcc
     7921 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccaccact gatttgagcg
     7981 tcagatttcg tgatgcttgt caggggggcg gagcctatgg aaaaacggct ttgccgcggc
     8041 cctctcactt ccctgttaag tatcttcctg gcatcttcca ggaaatctcc gccccgttcg
     8101 taagccattt ccgctcgccg cagtcgaacg accgagcgta gcgagtcagt gagcgaggaa
     8161 gcggaatata tcctgtatca catattctgc tgacgcaccg gtgcagcctt ttttctcctg
     8221 ccacatgaag cacttcactg acaccctcat cagtgccaac atagtaagcc agtatacact
     8281 ccgctagcgc tgatgtccgg cggtgctttt gccgttacgc accaccccgt catgagatta
     8341 cgcgttttag cggtgcgtac aattaaggga ttatggtaaa tccacttact gtctgccctc
     8401 gtagccatcg agataaaccg cagtactccg gccacgatgc gtccggcgta gaggatcgag
     8461 atct
//
