LOCUS       sequence-486828-        5029 bp    DNA     circular SYN 22-SEP-2025
DEFINITION  Express HaloTag-RBX1 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 5029)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5029)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 22, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: NanoBRET Assay Vector, HaloTag-RBX1
FEATURES             Location/Qualifiers
     source          1..5029
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      862..871
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(880..899)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(881..899)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(941..960)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     intron          943..1075
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        complement(1211..1414)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1240..1260)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1415..1794
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     misc_feature    1985..2268
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
     rep_origin      2384..2972
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2873..2892
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3130..3921,3922..3990))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3753..3772
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3991..4095)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    4342..4463
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4379..4398)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4433..4452
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4500..4518
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     regulatory      4948..4957
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(4971..4991)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             complement(join(5004..5029,1..865))
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
ORIGIN
        1 agcgcgcgat ctcgctgccg atcaggtccg ggttgtcttc ttgcagcaga ttcagacccg
       61 ggccgatgtc cacagccttg cagttaggca ggcttttggc caggcgagcg gcttcggccg
      121 gtgggatcag aacgcctggg gtgccccaga acagcagctt cgggacaggg gactggtgca
      181 gccagtccat gtattcttcg accagcgcga cgatgttcgc tggctcaccg gcgattggca
      241 gctcgtttgg gaagcgccac agtggctcgc ggtcaacagg attcaggaac ggctcgcggt
      301 aatggtccat ctcgacttca gtcagcgggc ggacgacacc catcggcagc gtaccctcga
      361 taaaaacgtt ctgatcgatg atcagcttgc ggccgacgtc ggtggtgcgg aaggcctgga
      421 aggtctcgcg ggcaaattct ggccattcgt cccaggtcgg gatagggcgg atgaactcca
      481 taaatgcaat acctttgacg cgctctggat tgcgcttggc ccagtggaaa cccagagcgg
      541 agccccagtc gtgaatgacc aggacgacct cttccagacc cagggcttcg atgaaggcat
      601 ccatgaagcg gacgtggtcg tcgaagaaat aacccaggtc tggtttgtcg gatttgccca
      661 taccgatcag gtctggagca atgcagcgat gggtcggtgc aacatgcggg atgatgttgc
      721 gccacacgta ggaggaggtc gggttaccgt gcaggaacag cacaggggtg ccatcgcgcg
      781 gaccaacatc gacgtagtgc atgcgctcgc ccaggacttc cacataatgg gggtcgaatg
      841 gaaagccagt accgatttct gccatggtgg ctttgctagc cctatagtga gtcgtattaa
      901 tactctagcc ttaagagctg taattgaact gggagtggac acctgtggag agaaaggcaa
      961 agtggatgtc agtaagacca ataggtgcct atcagaaacg caagagtctt ctctgtctcg
     1021 acaagcccag tttctattgg tctccttaaa cctgtcttgt aaccttgata cttacctgcc
     1081 cagtgcctca cgaccaactt ctgcagctta agttcgagac tgttgtgtca gaagcactga
     1141 ctgcgttagc aatttaactg tgataaacta ccgcaataaa gcttctagtg atctgacggt
     1201 tcactaaacc agctctgctt atatagacct cccaccgtac acgcctaccg cccatttgcg
     1261 tcaacggggc ggggttatta cgacattttg gaaagtcccg ttgattttgg tgccaaaaca
     1321 aactcccatt gacgtcaatg gggtggagac ttggaaatcc ccgtgagtca aaccgctatc
     1381 cacgcccatt ggtgtactgc caaaaccgca tcaccatggt aatagcgatg actaatacgt
     1441 agatgtactg ccaagtagga aagtcccgta aggtcatgta ctgggcataa tgccaggcgg
     1501 gccatttacc gtcattgacg tcaatagggg gcggacttgg catatgatac acttgatgta
     1561 ctgccaagtg ggcagtttac cgtaaatact ccacccattg acgtcaatgg aaagtcccta
     1621 ttggcgttac tatgggaaca tacgtcatta ttgacgtcaa tgggcggggg tcgttgggcg
     1681 gtcagccagg cgggccattt accgtaagtt atgtaacgcg gaactccata tatgggctat
     1741 gaactaatga ccccgtaatt gattactatt aataactagt caataatcaa tgccaacatg
     1801 gcggtcatat tggacatgag ccaatataaa tgtacatatt atgatataga tacaacgtat
     1861 gcaatggcca atagccaata ttgatttatg ctatataacc aatgaataat atggctaatg
     1921 gccaatattg aagatctcga tcctctacgc cggacgcatc gtggccggag tactgcggtt
     1981 tatctcgatg gctacgaggg cagacagtaa gtggatttac cataatccct taattgtacg
     2041 caccgctaaa acgcgttcag cgcgatcacg gcagcagaca ggtaaaaatg gcaacaaacc
     2101 acccgaaaaa ctgccgcaat cgcgcctgat aaattttaac cgtatgaata cctatgcaac
     2161 cagagggtac aggccacatt acccccactt aatccactga agctgccatt tttcatggtt
     2221 tcaccatccc agcgaagggc catccagcgt gcgttcctgt atttccggac tataaaagga
     2281 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     2341 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgaaat cctttttttc
     2401 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     2461 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     2521 caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     2581 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     2641 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     2701 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     2761 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     2821 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     2881 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     2941 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     3001 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     3061 tggataaccg tattaccgcc tttgagtgag ctgataccgg ttgggcgtcg cttggtcggt
     3121 catttcgaat taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     3181 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     3241 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     3301 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     3361 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     3421 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     3481 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     3541 caaaaaagcg gttagctcct tcggtcctcc gatagttgtc agaagtaagt tggccgcagt
     3601 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     3661 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     3721 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     3781 gaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     3841 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     3901 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     3961 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     4021 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     4081 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattac
     4141 gtagaaagcc agtccgcaga aacggtgctg accccggatg aatgtcagct actgggctat
     4201 ctggacaagg gaaaacgcaa gcgcaaagag gatccattag aattcgattt taccacattt
     4261 gtagaggttt tacttgctta aaaaaacctc ccacatctcc ccctgaacct gaaacataaa
     4321 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
     4381 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     4441 tccaaactca tcaatgtatc ttatcatgtc tgctcgaagc ggccgcccca aggggttatg
     4501 ctagttattg ctcagcggtg gcagcagcca actcagcttc ctttcgggct ttgttagcag
     4561 ccggatcagc ttgcatgcct gcaggtcgac tctagaggat ccccgggtac cgagcccgaa
     4621 ttcgtttaaa cgtgcccata cttttggaat tcccactctc tgttgtccaa tggacacacc
     4681 tgtcgtgttt tgagccagcg agagatgcag tggaagtgaa aagcatggtt acagactccc
     4741 catgcgacag tacactcttc tgaagtagcg gacgcctggt tagcttgaca ttctatgcaa
     4801 agatccataa tgtggttcct gcagatggca cagttatcaa ccacaatatc ccaggcccag
     4861 agggctactg cattccactt tttcacttca aagcgcttct tgcccgcgcc gctgttggtg
     4921 ccgctcgggg tatccacatc catcgctgcc gccatggcga tcgcgttatc gctctgaaag
     4981 tacagatcct cagtggttgg ctcgccggaa atctcgagcg tcgacagcc
//
