LOCUS       sequence-478504-        6217 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Express HaloTag-YAP 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 6217)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6217)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 7, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: HaloTag-YAP Fusion Vector
FEATURES             Location/Qualifiers
     source          1..6217
                     /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(3631..3649)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3686..3807
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3697..3716)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3751..3770
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4054..4158
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4159..4227,4228..5019)
                     /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(4377..4396)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(5177..5765)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5257..5276)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5881..6164
                     /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 tcgccatgga tcccgggcag cagccgccgc ctcaaccggc
     2041 cccccagggc caagggcagc cgccttcgca gcccccgcag gggcagggcc cgccgtccgg
     2101 acccgggcaa ccggcacccg cggcgaccca ggcggcgccg caggcacccc ccgccgggca
     2161 tcagatcgtg cacgtccgcg gggactcgga gaccgacctg gaggcgctct tcaacgccgt
     2221 catgaacccc aagacggcca acgtgcccca gaccgtgccc atgaggctcc ggaagctgcc
     2281 cgactccttc ttcaagccgc cggagcccaa atcccactcc cgacaggcca gtactgatgc
     2341 aggcactgca ggagccctga ctccacagca tgttcgagct cattcctctc cagcttctct
     2401 gcagttggga gctgtttctc ctgggacact gacccccact ggagtagtct ctggcccagc
     2461 agctacaccc acagctcagc atcttcgaca gtcttctttt gagatacctg atgatgtacc
     2521 tctgccagca ggttgggaga tggcaaagac atcttctggt cagagatact tcttaaatca
     2581 catcgatcag acaacaacat ggcaggaccc caggaaggcc atgctgtccc agatgaacgt
     2641 cacagccccc accagtccac cagtgcagca gaatatgatg aactcggctt caggtcctct
     2701 tcctgatgga tgggaacaag ccatgactca ggatggagaa atttactata taaaccataa
     2761 gaacaagacc acctcttggc tagacccaag gcttgaccct cgttttgcca tgaaccagag
     2821 aatcagtcag agtgctccag tgaaacagcc accacccctg gctccccaga gcccacaggg
     2881 aggcgtcatg ggtggcagca actccaacca gcagcaacag atgcgactgc agcaactgca
     2941 gatggagaag gagaggctgc ggctgaaaca gcaagaactg cttcggcagg caatgcggaa
     3001 tatcaatccc agcacagcaa attctccaaa atgtcaggag ttagccctgc gtagccagtt
     3061 accaacactg gagcaggatg gtgggactca aaatccagtg tcttctcccg ggatgtctca
     3121 ggaattgaga acaatgacga ccaatagctc agatcctttc cttaacagtg gcacctatca
     3181 ctctcgagat gagagtacag acagtggact aagcatgagc agctacagtg tccctcgaac
     3241 cccagatgac ttcctgaaca gtgtggatga gatggataca ggtgatacta tcaaccaaag
     3301 caccctgccc tcacagcaga accgtttccc agactacctt gaagccattc ctgggacaaa
     3361 tgtggacctt ggaacactgg aaggagatgg aatgaacata gaaggagagg agctgatgcc
     3421 aagtctgcag gaagctttga gttctgacat ccttaatgac atggagtctg ttttggctgc
     3481 caccaagcta gataaagaaa gctttcttac atggttagtt taaacgaatt cgggctcggt
     3541 acccggggat cctctagagt cgacctgcag gcatgcaagc tgatccggct gctaacaaag
     3601 cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg
     3661 gggcggccgc ttcgagcaga catgataaga tacattgatg agtttggaca aaccacaact
     3721 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
     3781 accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag
     3841 gttcaggggg agatgtggga ggttttttta agcaagtaaa acctctacaa atgtggtaaa
     3901 atcgaattct aatggatcct ctttgcgctt gcgttttccc ttgtccagat agcccagtag
     3961 ctgacattca tccggggtca gcaccgtttc tgcggactgg ctttctacgt aatggtttct
     4021 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     4081 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     4141 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     4201 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     4261 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     4321 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcactttcaa agttctgcta
     4381 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     4441 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     4501 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     4561 ttacttctga caactatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     4621 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     4681 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     4741 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     4801 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     4861 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     4921 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     4981 atcgctgaga taggtgcctc actgattaag cattggtaat tcgaaatgac cgaccaagcg
     5041 acgcccaacc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     5101 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     5161 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     5221 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     5281 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     5341 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     5401 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     5461 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     5521 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     5581 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     5641 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     5701 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     5761 ttcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     5821 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttatagt
     5881 ccggaaatac aggaacgcac gctggatggc ccttcgctgg gatggtgaaa ccatgaaaaa
     5941 tggcagcttc agtggattaa gtgggggtaa tgtggcctgt accctctggt tgcataggta
     6001 ttcatacggt taaaatttat caggcgcgat tgcggcagtt tttcgggtgg tttgttgcca
     6061 tttttacctg tctgctgccg tgatcgcgct gaacgcgttt tagcggtgcg tacaattaag
     6121 ggattatggt aaatccactt actgtctgcc ctcgtagcca tcgagataaa ccgcagtact
     6181 ccggccacga tgcgtccggc gtagaggatc gagatct
//
