LOCUS       sequence-475083-        6248 bp    DNA     circular SYN 21-JUL-2025
DEFINITION  Express HiBiT-RAF1 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 6248)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6248)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 21, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: HiBiT-RAF1 Fusion Vector
FEATURES             Location/Qualifiers
     source          1..6248
                     /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(3191..3209)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3246..3367
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3257..3276)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3311..3330
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3612..3969
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3612..3803
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3635..3655)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3820..3955
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3984..4031
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4063..4857
                     /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(4117..4136)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4727..4746
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4921..4969
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(5208..5796)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5288..5307)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5912..6195
                     /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 ggctagcgct caccatggtg agcggctggc
     1081 ggctgttcaa gaagattagc gggagctccg gtggctcgag cggtgcgatc gccatggagc
     1141 acatacaggg agcttggaag acgatcagca atggttttgg attcaaagat gccgtgtttg
     1201 atggctccag ctgcatctct cctacaatag ttcagcagtt tggctatcag cgccgggcat
     1261 cagatgatgg caaactcaca gatccttcta agacaagcaa cactatccgt gttttcttgc
     1321 cgaacaagca aagaacagtg gtcaatgtgc gaaatggaat gagcttgcat gactgcctta
     1381 tgaaagcact caaggtgagg ggcctgcaac cagagtgctg tgcagtgttc agacttctcc
     1441 acgaacacaa aggtaaaaaa gcacgcttag attggaatac tgatgctgcg tctttgattg
     1501 gagaagaact tcaagtagat ttcctggatc atgttcccct cacaacacac aactttgctc
     1561 ggaagacgtt cctgaagctt gccttctgtg acatctgtca gaaattcctg ctcaatggat
     1621 ttcgatgtca gacttgtggc tacaaatttc atgagcactg tagcaccaaa gtacctacta
     1681 tgtgtgtgga ctggagtaac atcagacaac tcttattgtt tccaaattcc actattggtg
     1741 atagtggagt cccagcacta ccttctttga ctatgcgtcg tatgcgagag tctgtttcca
     1801 ggatgcctgt tagttctcag cacagatatt ctacacctca cgccttcacc tttaacacct
     1861 ccagtccctc atctgaaggt tccctctccc agaggcagag gtcgacatcc acacctaatg
     1921 tccacatggt cagcaccacc ctgcctgtgg acagcaggat gattgaggat gcaattcgaa
     1981 gtcacagcga atcagcctca ccttcagccc tgtccagtag ccccaacaat ctgagcccaa
     2041 caggctggtc acagccgaaa acccccgtgc cagcacaaag agagcgggca ccagtatctg
     2101 ggacccagga gaaaaacaaa attaggcctc gtggacagag agattcaagc tattattggg
     2161 aaatagaagc cagtgaagtg atgctgtcca ctcggattgg gtcaggctct tttggaactg
     2221 tttataaggg taaatggcac ggagatgttg cagtaaagat cctaaaggtt gtcgacccaa
     2281 ccccagagca attccaggcc ttcaggaatg aggtggctgt tctgcgcaaa acacggcatg
     2341 tgaacattct gcttttcatg gggtacatga caaaggacaa cctggcaatt gtgacccagt
     2401 ggtgcgaggg cagcagcctc tacaaacacc tgcatgtcca ggagaccaag tttcagatgt
     2461 tccagctaat tgacattgcc cggcagacgg ctcagggaat ggactatttg catgcaaaga
     2521 acatcatcca tagagacatg aaatccaaca atatatttct ccatgaaggc ttaacagtga
     2581 aaattggaga ttttggtttg gcaacagtaa agtcacgctg gagtggttct cagcaggttg
     2641 aacaacctac tggctctgtc ctctggatgg ccccagaggt gatccgaatg caggataaca
     2701 acccattcag tttccagtcg gatgtctact cctatggcat cgtattgtat gaactgatga
     2761 cgggggagct tccttattct cacatcaaca accgagatca gatcatcttc atggtgggcc
     2821 gaggatatgc ctccccagat cttagtaagc tatataagaa ctgccccaaa gcaatgaaga
     2881 ggctggtagc tgactgtgtg aagaaagtaa aggaagagag gcctcttttt ccccagatcc
     2941 tgtcttccat tgagctgctc caacactctc taccgaagat caaccggagc gcttccgagc
     3001 catccttgca tcgggcagcc cacactgagg atatcaatgc ttgcacgctg accacgtccc
     3061 cgaggctgcc tgtcttcgtt taaacgaatt cgggctcggt acccggggat cctctagagt
     3121 cgacctgcag gcatgcaagc tgatccggct gctaacaaag cccgaaagga agctgagttg
     3181 gctgctgcca ccgctgagca ataactagca taaccccttg gggcggccgc ttcgagcaga
     3241 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     3301 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     3361 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg agatgtggga
     3421 ggttttttta agcaagtaaa acctctacaa atgtggtaaa atcgaatttt aacaaaatat
     3481 taacgcttac aatttcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     3541 ccgcatacgc ggatctgcgc agcaccatgg cctgaaataa cctctgaaag aggaacttgg
     3601 ttaggtacct tctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga
     3661 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3721 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3781 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     3841 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     3901 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     3961 ttttgcaaaa agcttaatta actgttgaca attaatcatc ggcatagtat atcggcatag
     4021 tataatacga caaggtgagg aactaaaccc aggaggcaga tcatgattga acaagatgga
     4081 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     4141 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     4201 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     4261 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     4321 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     4381 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     4441 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     4501 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     4561 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     4621 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     4681 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     4741 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     4801 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     4861 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgatggccgc
     4921 aataaaatat ctttattttc attacatctg tgtgttggtt ttttgtgtga atcgatagcg
     4981 ataaggatcc tctttgcgct tgcgttttcc cttgtccaga tagcccagta gctgacattc
     5041 atccggggtc agcaccgttt ctgcggactg gctttctacc cggtatcagc tcactcaaag
     5101 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     5161 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     5221 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     5281 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     5341 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     5401 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     5461 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     5521 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     5581 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     5641 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     5701 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     5761 aagcagcaga ttacgcgcag aaaaaaagga tttcaagaag atcctttgat cttttctacg
     5821 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     5881 aaaaggatct tcacctagat ccttttatag tccggaaata caggaacgca cgctggatgg
     5941 cccttcgctg ggatggtgaa accatgaaaa atggcagctt cagtggatta agtgggggta
     6001 atgtggcctg taccctctgg ttgcataggt attcatacgg ttaaaattta tcaggcgcga
     6061 ttgcggcagt ttttcgggtg gtttgttgcc atttttacct gtctgctgcc gtgatcgcgc
     6121 tgaacgcgtt ttagcggtgc gtacaattaa gggattatgg taaatccact tactgtctgc
     6181 cctcgtagcc atcgagataa accgcagtac tccggccacg atgcgtccgg cgtagaggat
     6241 cgagatct
//
