LOCUS       sequence-472400-        6456 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express TEAD4-HaloTag(R) 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 6456)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6456)
  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            "Unpublished"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: TEAD4-HaloTag Vector
FEATURES             Location/Qualifiers
     source          1..6456
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(747..1541)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(858..877)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1468..1487
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(1573..1620)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     protein_bind    1630..1663
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    1778..1899
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     1869..1888
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(1986..2870)
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="EIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYV
                     WRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHD
                     WGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQ
                     NVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEE
                     YMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIG
                     SEIARWLSTLEISG"
     CDS             complement(2886..2906)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     primer_bind     complement(4242..4261)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4243..4261)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4303..4322)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     intron          4305..4437
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        complement(4573..4776)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4602..4622)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4777..5080
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    5313..5394
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        5473..5830
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5473..5664
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5496..5516)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5681..5816
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      5855..5864
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5861..6259
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    6283..6331
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 ttgaaatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccggttg
      721 ggcgtcgctt ggtcggtcat ttcgaatcag aagaactcgt caagaaggcg atagaaggcg
      781 atgcgctgcg aatcgggagc ggcgataccg taaagcacga ggaagcggtc agcccattcg
      841 ccgccaagct cttcagcaat atcacgggta gccaacgcta tgtcctgata gcggtccgcc
      901 acacccagcc ggccacagtc gatgaatcca gaaaagcggc cattttccac catgatattc
      961 ggcaagcagg catcgccatg ggtcacgacg agatcctcgc cgtcgggcat gcgcgccttg
     1021 agcctggcga acagttcggc tggcgcgagc ccctgatgct cttcgtccag atcatcctga
     1081 tcgacaagac cggcttccat ccgagtacgt gctcgctcga tgcgatgttt cgcttggtgg
     1141 tcgaatgggc aggtagccgg atcaagcgta tgcagccgcc gcattgcatc agccatgatg
     1201 gatactttct cggcaggagc aaggtgagat gacaggagat cctgccccgg cacttcgccc
     1261 aatagcagcc agtcccttcc cgcttcagtg acaacgtcga gcacagctgc gcaaggaacg
     1321 cccgtcgtgg ccagccacga tagccgcgct gcctcgtcct gcagttcatt cagggcaccg
     1381 gacaggtcgg tcttgacaaa aagaaccggg cgcccctgcg ctgacagccg gaacacggcg
     1441 gcatcagagc agccgattgt ctgttgtgcc cagtcatagc cgaatagcct ctccacccaa
     1501 gcggccggag aacctgcgtg caatccatct tgttcaatca tgatctgcct cctgggttta
     1561 gttcctcacc ttgtcgtatt atactatgcc gatatactat gccgatgatt aattgtcaac
     1621 agtggatcca taacttcgta taatgtatgc tatacgaagt tatattagaa ttcttaatta
     1681 agattttacc acatttgtag aggttttact tgcttaaaaa aacctcccac atctccccct
     1741 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa
     1801 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     1861 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgct cgaagcggcc
     1921 ttgctcagcg gtggcagcag ccaaatcagc ttgcatgcct gcaggtcgac tctagaattc
     1981 tattaaccgg aaatctccag agtagacagc cagcgcgcga tctcgctgcc gatcaggtcc
     2041 gggttgtctt cttgcagcag attcagaccc gggccgatgt ccacagcctt gcagttaggc
     2101 aggcttttgg ccaggcgagc ggcttcggcc ggtgggatca gaacgcctgg ggtgccccag
     2161 aacagcagct tcgggacagg ggactggtgc agccagtcca tgtattcttc gaccagcgcg
     2221 acgatgttcg ctggctcacc ggcgattggc agctcgtttg ggaagcgcca cagtggctcg
     2281 cggtcaacag gattcaggaa cggctcgcgg taatggtcca tctcgacttc agtcagcggg
     2341 cggacgacac ccatcggcag cgtaccctcg ataaaaacgt tctgatcgat gatcagcttg
     2401 cggccgacgt cggtggtgcg gaaggcctgg aaggtctcgc gggcaaattc tggccattcg
     2461 tcccaggtcg ggatagggcg gatgaactcc ataaatgcaa tacctttgac gcgctctgga
     2521 ttgcgcttgg cccagtggaa acccagagcg gagccccagt cgtgaatgac caggacgacc
     2581 tcttccagac ccagggcttc gatgaaggca tccatgaagc ggacgtggtc gtcgaagaaa
     2641 taacccaggt ctggtttgtc ggatttgccc ataccgatca ggtctggagc aatgcagcga
     2701 tgggtcggtg caacatgcgg gatgatgttg cgccacacgt aggaggaggt cgggttaccg
     2761 tgcaggaaca gcacaggggt gccatcgcgc ggaccaacat cgacgtagtg catgcgctcg
     2821 cccaggactt ccacataatg ggggtcgaat ggaaagccag taccgatttc ggatccatcg
     2881 ttatcgctct gaaagtacag atcctcagtg gttggctcga gagaaacttc tttcaccagc
     2941 ctgtagatgt ggtgctgagc cccgtgctca ctggctgaca cctcaaagac ataggcaatg
     3001 cacagcaagg tctcctgtgt gtctctgttg gtgaccacct gcaggatggt gaagttctcc
     3061 agcacgctgt tcatcatgta cttctcaggg aggtgcttga gcttgtggat gaagttgatc
     3121 atgtactcac agagcgggga ccggtggatg cggtaagagt agtgtccatt ctcatagcga
     3181 gcatactctg tctcaacttt ctccaccacc tgcttgccga aagagcagac cttcgtggag
     3241 caggtgatga tcatgttctc ggggctctca tactggctgg agaccccata gaaggagctg
     3301 ccttcatcct cgatgttggt gttgaggtct gcccagaact tcacaagaaa aaaggcattg
     3361 gagggtcccc gttcgaagag atccttgagt ccaccctttt tctccgggaa tttgtcatag
     3421 atttggcgga tgtccacggc ttcgaggtag gggtcgctgt agcttgggct ggactggcca
     3481 atgtgcacga acaggtgctt gttgtacgtg tccgggtcct gctgctgctc caggaaggca
     3541 gagaactcca acatccagag cttggagctg gccacgctgc ggccctgcca tgggggtgcc
     3601 gggggcgcag agggcgatgg ggcgggccct gcaggagact caaaccctgg cagaggcagc
     3661 ggaggctgga cagcataggt ttgctgagag aaaggcttca catcatggga cgttccggct
     3721 tggcctggca aagctccttg ccaaaaccct gagactgctg ggcggccggg gccccgggcg
     3781 agggccatgc tactgtggaa ggccgtggcg gagatgatct gtgcagacga catggcagcc
     3841 atgctctgca gggccttgtc cttagctgcc tggtccttta gcttggcctg gatctcgcga
     3901 gctttgcgac gagccagcac ctggatgtgg ctggagacct gcttcctggt gcgggtcttc
     3961 cctgtccgga gcttgatgta gcgggcaatc agctcgttcc gaccatacat cttgccctcg
     4021 tccgacagga tgattttgcg cctgccacag ggcgggtaga tggcgagggc ctcctggaaa
     4081 ctctgctcaa tatccgggct ccacacgccc tctgcgtcat tgtcgatggg cttgtccagt
     4141 gcctgactgc ccccagaggc ggtgctcccc tcaggggagg tgggagagct ccactcgttg
     4201 gaggtaatgg tgccggccgt gccctccatg gcgatcgcta gccctatagt gagtcgtatt
     4261 aatactctag ccttaagagc tgtaattgaa ctgggagtgg acacctgtgg agagaaaggc
     4321 aaagtggatg tcagtaagac caataggtgc ctatcagaaa cgcaagagtc ttctctgtct
     4381 cgacaagccc agtttctatt ggtctcctta aacctgtctt gtaaccttga tacttacctg
     4441 cccagtgcct cacgaccaac ttctgcagct taagttcgag actgttgtgt cagaagcact
     4501 gactgcgtta gcaatttaac tgtgataaac taccgcaata aagcttctag tgatctgacg
     4561 gttcactaaa ccagctctgc ttatatagac ctcccaccgt acacgcctac cgcccatttg
     4621 cgtcaacggg gcggggttat tacgacattt tggaaagtcc cgttgatttt ggtgccaaaa
     4681 caaactccca ttgacgtcaa tggggtggag acttggaaat ccccgtgagt caaaccgcta
     4741 tccacgccca ttggtgtact gccaaaaccg catcaccatg gtaatagcga tgactaatac
     4801 gtagatgtac tgccaagtag gaaagtcccg taaggtcatg tactgggcat aatgccaggc
     4861 gggccattta ccgtcattga cgtcaatagg gggcggactt ggcatatgat acacttgatg
     4921 tactgccaag tgggcagttt accgtaaata ctccacccat tgacgtcaat ggaaagtccc
     4981 tattggcgtt actatgggaa catacgtcat tattgacgtc aatgggcggg ggtcgttggg
     5041 cggtcagcca ggcgggccat ttaccgtaag ttatgtaacg cggaactcca tatatgggct
     5101 atgaactaat gaccccgtaa ttgattacta ttaataacta gtggcgcgcc ataatcctag
     5161 gctctcgtac gaagtcagct gataacgtcg acgtattgcg gccgcggatc ataatcagcc
     5221 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
     5281 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
     5341 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgccgacgt
     5401 cttaattaac gcgtcttgcg cagcaccatg gcctgaaata acctctgaaa gaggaacttg
     5461 gttaggtacc ttctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg
     5521 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     5581 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     5641 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     5701 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     5761 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     5821 cttttgcaaa aagctcgatt cttctgacac tagcgccacc atggccaagc ctttgtctca
     5881 agaagaatcc accctcattg aaagagcaac ggctacaatc aacagcatcc ccatctctga
     5941 agactacagc gtcgccagcg cagctctctc tagcgacggc cgcatcttca ctggtgtcaa
     6001 tgtatatcat tttactgggg gaccttgtgc agaactcgtg gtgctgggca ctgctgctgc
     6061 tgcggcagct ggcaacctga cttgtatcgt cgcgatcgga aatgagaaca ggggcatctt
     6121 gagcccctgc ggacggtgcc gacaggtgct tctcgatctg catcctggga tcaaagccat
     6181 agtgaaggac agtgatggac agccgacggc agttgggatt cgtgaattgc tgccctctgg
     6241 ttatgtgtgg gagggctaaa ctctagccat cacgatggcc gcaataaaat atctttattt
     6301 tcattacatc tgtgtgttgg ttttttgtgt gacgattccg gactataaaa ggatctaggt
     6361 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     6421 agcgtcagac cccgtagaaa agatcaaagg atcttc
//
