LOCUS       sequence-479215-        6610 bp    DNA     circular SYN 25-JUL-2025
DEFINITION  Express HaloTag-Syk 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 6610)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6610)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 25, 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            "Unpublished"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-Syk
FEATURES             Location/Qualifiers
     source          1..6610
                     /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(4024..4042)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    4079..4200
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4090..4109)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4144..4163
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4447..4551
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4552..4620,4621..5412)
                     /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(4770..4789)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(5570..6158)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5650..5669)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6274..6557
                     /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 cagcagcggc atggctgaca gcgccaacca
     2041 cctgcccttc tttttcggca acatcacccg ggaggaggca gaagattacc tggtccaggg
     2101 gggcatgagt gatgggcttt atttgctgcg ccagagccgc aactacctgg gtggcttcgc
     2161 cctgtccgtg gcccacggga ggaaggcaca ccactacacc atcgagcggg agctgaatgg
     2221 cacctacgcc atcgccggtg gcaggaccca tgccagcccc gccgacctct gccactacca
     2281 ctcccaggag tctgatggcc tggtctgcct cctcaagaag cccttcaacc ggccccaagg
     2341 ggtgcagccc aagactgggc cctttgagga tttgaaggaa aacctcatca gggaatatgt
     2401 gaagcagaca tggaacctgc agggtcaggc tctggagcag gccatcatca gtcagaagcc
     2461 tcagctggag aagctgatcg ctaccacagc ccatgaaaaa atgccttggt tccatggaaa
     2521 aatctctcgg gaagaatctg agcaaattgt cctgatagga tcaaagacaa atggaaagtt
     2581 cctgatccga gccagagaca acaacggctc ctacgccctg tgcctgctgc acgaagggaa
     2641 ggtgctgcac tatcgcatcg acaaagacaa gacagggaag ctctccatcc ccgagggaaa
     2701 gaagttcgac acgctctggc agctagtcga gcattattct tataaagcag atggtttgtt
     2761 aagagttctt actgtcccat gtcaaaaaat cggcacacag ggaaatgtta attttggagg
     2821 ccgtccacaa cttccaggtt cccatcctgc gacttggtca gcgggtggaa taatctcaag
     2881 aatcaaatca tactccttcc caaagcctgg ccacagaaag tcctcccctg cccaagggaa
     2941 ccggcaagag agtactgtgt cattcaatcc gtatgagcca gaacttgcac cctgggctgc
     3001 agacaaaggc ccccagagag aagccctacc catggacaca gaggtgtacg agagccccta
     3061 cgcggacccc gaggagatca ggcccaagga ggtttacctg gaccgaaagc tgctgacgct
     3121 ggaagacaaa gaactgggct ctggtaattt tggaactgtg aaaaagggct actaccaaat
     3181 gaaaaaagtt gtgaaaaccg tggctgtgaa aatactgaaa aacgaggcca atgaccccgc
     3241 tcttaaagat gagttattag cagaagcaaa tgtcatgcag cagctggaca acccgtacat
     3301 cgtgcggatg atcgggatat gcgaggccga gtcctggatg ctggttatgg agatggcaga
     3361 acttggtccc ctcaataagt atttgcagca gaacagacat gtcaaggata agaacatcat
     3421 agaactggtt catcaggttt ccatgggcat gaagtacttg gaggagagca attttgtgca
     3481 cagagatctg gctgcaagaa atgtgttgct agttacccaa cattacgcca agatcagtga
     3541 tttcggactt tccaaagcac tgcgtgctga tgaaaactac tacaaggccc agacccatgg
     3601 aaagtggcct gtcaagtggt acgctccgga atgcatcaac tactacaagt tctccagcaa
     3661 aagcgatgtc tggagctttg gagtgttgat gtgggaagca ttctcctatg ggcagaagcc
     3721 atatcgaggg atgaaaggaa gtgaagtcac cgctatgtta gagaaaggag agcggatggg
     3781 gtgccctgca gggtgtccaa gagagatgta cgatctcatg aatctgtgct ggacatacga
     3841 tgtggaaaac aggcccggat tcgcagcagt ggaactgcgg ctgcgcaatt actactatga
     3901 cgtggtgaac gtttaaacga attcgggctc ggtacccggg gatcctctag agtcgacctg
     3961 caggcatgca agctgatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg
     4021 ccaccgctga gcaataacta gcataacccc ttggggcggc cgcttcgagc agacatgata
     4081 agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt
     4141 tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt
     4201 aacaacaaca attgcattca ttttatgttt caggttcagg gggagatgtg ggaggttttt
     4261 ttaagcaagt aaaacctcta caaatgtggt aaaatcgaat tctaatggat cctctttgcg
     4321 cttgcgtttt cccttgtcca gatagcccag tagctgacat tcatccgggg tcagcaccgt
     4381 ttctgcggac tggctttcta cgtaatggtt tcttagacgt caggtggcac ttttcgggga
     4441 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     4501 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     4561 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     4621 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     4681 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     4741 tttccaatga tgagcacttt caaagttctg ctatgtggcg cggtattatc ccgtattgac
     4801 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     4861 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     4921 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaactat cggaggaccg
     4981 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     5041 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     5101 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     5161 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     5221 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     5281 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     5341 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     5401 aagcattggt aattcgaaat gaccgaccaa gcgacgccca accggtatca gctcactcaa
     5461 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     5521 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     5581 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     5641 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     5701 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     5761 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     5821 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     5881 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     5941 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     6001 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     6061 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     6121 gcaagcagca gattacgcgc agaaaaaaag gatttcaaga agatcctttg atcttttcta
     6181 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     6241 caaaaaggat cttcacctag atccttttat agtccggaaa tacaggaacg cacgctggat
     6301 ggcccttcgc tgggatggtg aaaccatgaa aaatggcagc ttcagtggat taagtggggg
     6361 taatgtggcc tgtaccctct ggttgcatag gtattcatac ggttaaaatt tatcaggcgc
     6421 gattgcggca gtttttcggg tggtttgttg ccatttttac ctgtctgctg ccgtgatcgc
     6481 gctgaacgcg ttttagcggt gcgtacaatt aagggattat ggtaaatcca cttactgtct
     6541 gccctcgtag ccatcgagat aaaccgcagt actccggcca cgatgcgtcc ggcgtagagg
     6601 atcgagatct
//
