LOCUS       sequence-472735-        6922 bp    DNA     circular SYN 24-JUN-2025
DEFINITION  Express HaloTag(R)-Suz12 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 6922)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 6922)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 24, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: NanoBRET Assay Vector, Suz12
FEATURES             Location/Qualifiers
     source          1..6922
                     /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"
     regulatory      1061..1070
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1067..1957
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
     CDS             1970..1990
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     primer_bind     complement(4336..4354)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    4391..4512
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4402..4421)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4456..4475
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4759..4863
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4864..4932,4933..5724)
                     /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     complement(5082..5101)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(5882..6470)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5962..5981)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6586..6869
                     /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 ggctagcaaa gccaccatgg cagaaatcgg
     1081 tactggcttt ccattcgacc cccattatgt ggaagtcctg ggcgagcgca tgcactacgt
     1141 cgatgttggt ccgcgcgatg gcacccctgt gctgttcctg cacggtaacc cgacctcctc
     1201 ctacgtgtgg cgcaacatca tcccgcatgt tgcaccgacc catcgctgca ttgctccaga
     1261 cctgatcggt atgggcaaat ccgacaaacc agacctgggt tatttcttcg acgaccacgt
     1321 ccgcttcatg gatgccttca tcgaagccct gggtctggaa gaggtcgtcc tggtcattca
     1381 cgactggggc tccgctctgg gtttccactg ggccaagcgc aatccagagc gcgtcaaagg
     1441 tattgcattt atggagttca tccgccctat cccgacctgg gacgaatggc cagaatttgc
     1501 ccgcgagacc ttccaggcct tccgcaccac cgacgtcggc cgcaagctga tcatcgatca
     1561 gaacgttttt atcgagggta cgctgccgat gggtgtcgtc cgcccgctga ctgaagtcga
     1621 gatggaccat taccgcgagc cgttcctgaa tcctgttgac cgcgagccac tgtggcgctt
     1681 cccaaacgag ctgccaatcg ccggtgagcc agcgaacatc gtcgcgctgg tcgaagaata
     1741 catggactgg ctgcaccagt cccctgtccc gaagctgctg ttctggggca ccccaggcgt
     1801 tctgatccca ccggccgaag ccgctcgcct ggccaaaagc ctgcctaact gcaaggctgt
     1861 ggacatcggc ccgggtctga atctgctgca agaagacaac ccggacctga tcggcagcga
     1921 gatcgcgcgc tggctgtcga cgctcgagat ttccggcgag ccaaccactg aggatctgta
     1981 ctttcagagc gataacgcga tcgccatggc gcctcagaag cacggcggtg ggggaggggg
     2041 cggctcgggg cccagcgcgg ggtccggggg aggcggcttc gggggttcgg cggcggtggc
     2101 ggcggcgacg gcttcgggcg gcaaatccgg cggcgggagc tgtggagggg gtggcagtta
     2161 ctcggcctcc tcctcctcct ccgcggcggc agcggcgggg gctgcggtgt taccggtgaa
     2221 gaagccgaaa atggagcacg tccaggctga ccacgagctt ttcctccagg cctttgagaa
     2281 gccaacacag atctatagat ttcttcgaac tcggaatctc atagcaccaa tatttttgca
     2341 cagaactctt acttacatgt ctcatcgaaa ctccagaaca aacatcaaaa ggaaaacatt
     2401 taaagttgat gatatgttat caaaagtaga gaaaatgaaa ggagagcaag aatctcatag
     2461 cttgtcagct catttgcagc ttacgtttac tggtttcttc cacaaaaatg ataagccatc
     2521 accaaactca gaaaatgaac aaaattctgt taccctggaa gtcctgcttg tgaaagtttg
     2581 ccacaaaaaa agaaaggatg taagttgtcc aataaggcaa gttcccacag gtaaaaagca
     2641 ggtgcctttg aatcctgacc tcaatcaaac aaaacccgga aatttcccgt cccttgcagt
     2701 ttccagtaat gaatttgaac ctagtaacag ccatatggtg aagtcttact cgttgctatt
     2761 tagagtgact cgtccaggaa gaagagagtt taatggaatg attaatggag aaaccaatga
     2821 aaatattgat gtcaatgaag agcttccagc cagaagaaaa cgaaatcgtg aggatgggga
     2881 aaagacattt gttgcacaaa tgacagtatt tgataaaaac aggcgcttac agcttttaga
     2941 tggggaatat gaagtagcca tgcaggaaat ggaagaatgt ccaataagca agaaaagagc
     3001 aacatgggag actattcttg atgggaagag gctgcctcca ttcgaaacat tttctcaggg
     3061 acctacgttg cagttcactc ttcgttggac aggagagacc aatgataaat ctacggctcc
     3121 tattgccaaa cctcttgcca ctagaaattc agagagtctc catcaggaaa acaagcctgg
     3181 ttcagttaaa cctactcaaa ctattgctgt taaagaatca ttgactacag atctacaaac
     3241 aagaaaagaa aaggatactc caaatgaaaa ccgacaaaaa ttaagaatat tttatcagtt
     3301 tctctataac aacaatacaa ggcaacaaac tgaagcaaga gatgacctgc attgcccttg
     3361 gtgtactctg aactgccgca aactttatag tttactcaag catcttaaac tctgccatag
     3421 cagatttatc ttcaactatg tttatcatcc aaaaggtgct aggatagatg tttctatcaa
     3481 tgagtgttat gatggctcct atgcaggaaa tcctcaggat attcatcgcc aacctggatt
     3541 tgcttttagt cgcaacggac cagttaagag aacacctatc acacatattc ttgtgtgcag
     3601 gccaaaacga acaaaagcaa gcatgtctga atttcttgaa tctgaagatg gggaagtaga
     3661 acagcaaaga acatatagta gtggccacaa tcgtctgtat ttccatagtg atacctgctt
     3721 acctctccgt ccacaagaaa tggaagtaga tagtgaagat gaaaaggatc ctgaatggct
     3781 aagagaaaaa accattacac aaattgaaga gttttctgat gttaatgaag gagagaaaga
     3841 agtgatgaaa ctctggaatc tccatgtcat gaagcatggg tttattgctg acaatcaaat
     3901 gaatcatgcc tgtatgctgt ttgtagaaaa ttatggacag aaaataatta agaagaattt
     3961 atgtcgaaac ttcatgcttc atctagtcag catgcatgac tttaatctta ttagcataat
     4021 gtcaatagat aaagctgtta ccaagctccg tgaaatgcag caaaaattag aaaaggggga
     4081 atctgcttcc cctgcaaacg aagaaataac tgaagaacaa aatgggacag caaatggatt
     4141 tagtgaaatt aactcaaaag agaaagcttt ggaaacagat agtgtctcag gggtttcaaa
     4201 acagagcaaa aaacaaaaac tcgtttaaac gaattcgggc tcggtacccg gggatcctct
     4261 agagtcgacc tgcaggcatg caagctgatc cggctgctaa caaagcccga aaggaagctg
     4321 agttggctgc tgccaccgct gagcaataac tagcataacc ccttggggcg gccgcttcga
     4381 gcagacatga taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa
     4441 aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc
     4501 aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggagatg
     4561 tgggaggttt ttttaagcaa gtaaaacctc tacaaatgtg gtaaaatcga attctaatgg
     4621 atcctctttg cgcttgcgtt ttcccttgtc cagatagccc agtagctgac attcatccgg
     4681 ggtcagcacc gtttctgcgg actggctttc tacgtaatgg tttcttagac gtcaggtggc
     4741 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     4801 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     4861 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     4921 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     4981 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     5041 cccgaagaac gttttccaat gatgagcact ttcaaagttc tgctatgtgg cgcggtatta
     5101 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     5161 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     5221 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaact
     5281 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     5341 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     5401 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     5461 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     5521 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     5581 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     5641 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     5701 gcctcactga ttaagcattg gtaattcgaa atgaccgacc aagcgacgcc caaccggtat
     5761 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     5821 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     5881 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     5941 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     6001 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     6061 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     6121 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     6181 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     6241 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     6301 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     6361 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     6421 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatttcaa gaagatcctt
     6481 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     6541 tcatgagatt atcaaaaagg atcttcacct agatcctttt atagtccgga aatacaggaa
     6601 cgcacgctgg atggcccttc gctgggatgg tgaaaccatg aaaaatggca gcttcagtgg
     6661 attaagtggg ggtaatgtgg cctgtaccct ctggttgcat aggtattcat acggttaaaa
     6721 tttatcaggc gcgattgcgg cagtttttcg ggtggtttgt tgccattttt acctgtctgc
     6781 tgccgtgatc gcgctgaacg cgttttagcg gtgcgtacaa ttaagggatt atggtaaatc
     6841 cacttactgt ctgccctcgt agccatcgag ataaaccgca gtactccggc cacgatgcgt
     6901 ccggcgtaga ggatcgagat ct
//
