LOCUS       sequence-473387-        4260 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express HaloTag(R)-MAFG 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 4260)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4260)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: HaloTag-MAFG-pFN24A Vector
FEATURES             Location/Qualifiers
     source          1..4260
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..45
                     /label=CMVd3 promoter
                     /note="truncated human cytomegalovirus (CMV) immediate 
                     early promoter that yields strongly reduced expression 
                     relative to the full-length promoter"
     primer_bind     77..96
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        77..95
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        101..119
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     101..118
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             136..1026
                     /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             1039..1059
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     primer_bind     complement(1674..1692)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    1729..1850
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1740..1759)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1794..1813
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2097..2201
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2202..2270,2271..3062)
                     /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(2420..2439)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(3220..3808)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3300..3319)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    3924..4207
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
ORIGIN
        1 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt
       61 cagatcacta gaagcttaat acgactcact atagggtacg atttaggtga cactatagaa
      121 taaggagaat tcatcatggc agaaatcggt actggctttc cattcgaccc ccattatgtg
      181 gaagtcctgg gcgagcgcat gcactacgtc gatgttggtc cgcgcgatgg cacccctgtg
      241 ctgttcctgc acggtaaccc gacctcctcc tacgtgtggc gcaacatcat cccgcatgtt
      301 gcaccgaccc atcgctgcat tgctccagac ctgatcggta tgggcaaatc cgacaaacca
      361 gacctgggtt atttcttcga cgaccacgtc cgcttcatgg atgccttcat cgaagccctg
      421 ggtctggaag aggtcgtcct ggtcattcac gactggggct ccgctctggg tttccactgg
      481 gccaagcgca atccagagcg cgtcaaaggt attgcattta tggagttcat ccgccctatc
      541 ccgacctggg acgaatggcc agaatttgcc cgcgagacct tccaggcctt ccgcaccacc
      601 gacgtcggcc gcaagctgat catcgatcag aacgttttta tcgagggtac gctgccgatg
      661 ggtgtcgtcc gcccgctgac tgaagtcgag atggaccatt accgcgagcc gttcctgaat
      721 cctgttgacc gcgagccact gtggcgcttc ccaaacgagc tgccaatcgc cggtgagcca
      781 gcgaacatcg tcgcgctggt cgaagaatac atggactggc tgcaccagtc ccctgtcccg
      841 aagctgctgt tctggggcac cccaggcgtt ctgatcccac cggccgaagc cgctcgcctg
      901 gccaaaagcc tgcctaactg caaggctgtg gacatcggcc cgggtctgaa tctgctgcaa
      961 gaagacaacc cggacctgat cggcagcgag atcgcgcgct ggctgtcgac gctcgagatt
     1021 tccggcgagc caaccactga ggatctgtac tttcagagcg ataacgcgat cgccatgacg
     1081 acccccaata aaggaaacaa ggccttgaag gtgaagcggg agccgggtga gaatggcacc
     1141 agcctgacgg atgaggagct ggtgaccatg tcggtgcggg agctgaacca gcacctgcgg
     1201 ggcctgtcca aggaggagat cgtccagctg aagcagcgcc ggcgcacgct caagaaccgc
     1261 ggctacgctg ccagctgccg cgtgaagcgg gtgacgcaga aggaggagct ggagaagcag
     1321 aaggcggagc tgcagcagga ggtggagaag ctggcctcag agaacgccag catgaagctg
     1381 gagctcgacg cgctgcgctc caagtacgag gcgctgcaga ccttcgcccg gacggtggcc
     1441 cgcagccccg tggcgccagc ccggggcccc cttgccgccg gcctggggcc cctcgtccca
     1501 ggcaaggtgg ccgccaccag cgtcatcaca atagtaaagt ccaagacgga tgcccgatcg
     1561 gtttaaacga attcgggctc ggtacccggg gatcctctag agtcgacctg caggcatgca
     1621 agctgatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga
     1681 gcaataacta gcataacccc ttggggcggc cgcttcgagc agacatgata agatacattg
     1741 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
     1801 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
     1861 attgcattca ttttatgttt caggttcagg gggagatgtg ggaggttttt ttaagcaagt
     1921 aaaacctcta caaatgtggt aaaatcgaat tctaatggat cctctttgcg cttgcgtttt
     1981 cccttgtcca gatagcccag tagctgacat tcatccgggg tcagcaccgt ttctgcggac
     2041 tggctttcta cgtaatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     2101 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     2161 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     2221 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     2281 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     2341 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     2401 tgagcacttt caaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     2461 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     2521 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     2581 tgagtgataa cactgcggcc aacttacttc tgacaactat cggaggaccg aaggagctaa
     2641 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     2701 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     2761 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     2821 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     2881 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     2941 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     3001 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     3061 aattcgaaat gaccgaccaa gcgacgccca accggtatca gctcactcaa aggcggtaat
     3121 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     3181 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     3241 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     3301 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     3361 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     3421 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     3481 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     3541 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     3601 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     3661 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     3721 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     3781 gattacgcgc agaaaaaaag gatttcaaga agatcctttg atcttttcta cggggtctga
     3841 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     3901 cttcacctag atccttttat agtccggaaa tacaggaacg cacgctggat ggcccttcgc
     3961 tgggatggtg aaaccatgaa aaatggcagc ttcagtggat taagtggggg taatgtggcc
     4021 tgtaccctct ggttgcatag gtattcatac ggttaaaatt tatcaggcgc gattgcggca
     4081 gtttttcggg tggtttgttg ccatttttac ctgtctgctg ccgtgatcgc gctgaacgcg
     4141 ttttagcggt gcgtacaatt aagggattat ggtaaatcca cttactgtct gccctcgtag
     4201 ccatcgagat aaaccgcagt actccggcca cgatgcgtcc ggcgtagagg atcgagatct
//
