LOCUS       sequence-473405-        8185 bp    DNA     circular SYN 14-JUL-2025
DEFINITION  Express TEAD1-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 8185)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 8185)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: TEAD1-HaloTag Vector
FEATURES             Location/Qualifiers
     source          1..8185
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    24..105
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     enhancer        338..641
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        642..845
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     796..816
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          981..1113
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1096..1115
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1157..1176
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1157..1175
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2488..2508
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             2524..3408
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="EIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYV
                     WRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHD
                     WGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQ
                     NVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEE
                     YMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIG
                     SEIARWLSTLEISG"
     polyA_signal    3495..3616
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3506..3525)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(3731..3764)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3774..3821
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3853..4647
                     /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(3907..3926)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4517..4536
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4805..5393)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4885..4904)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5519..5567
                     /note="synthetic polyadenylation signal"
     CDS             complement(5591..6628)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             5604..5612
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     promoter        complement(7007..8185)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          7016..7954
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     complement(7039..7059)
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
ORIGIN
        1 aagacgcgtt aattaagacg tcggcagtga aaaaaatgct ttatttgtga aatttgtgat
       61 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
      121 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac
      181 ctctacaaat gtggtatggc tgattatgat ccgcggccgc aatacgtcga cgttatcagc
      241 tgacttcgta cgagagccta ggattatggc gcgccactag ttattaatag taatcaatta
      301 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      361 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      421 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      481 ctgcccactt ggcagtacat caagtgtatc atatgccaag tccgccccct attgacgtca
      541 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttacgg gactttccta
      601 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      661 acaccaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      721 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaata
      781 accccgcccc gttgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      841 gagctggttt agtgaaccgt cagatcacta gaagctttat tgcggtagtt tatcacagtt
      901 aaattgctaa cgcagtcagt gcttctgaca caacagtctc gaacttaagc tgcagaagtt
      961 ggtcgtgagg cactgggcag gtaagtatca aggttacaag acaggtttaa ggagaccaat
     1021 agaaactggg cttgtcgaga cagagaagac tcttgcgttt ctgataggca cctattggtc
     1081 ttactgacat ccactttgcc tttctctcca caggtgtcca ctcccagttc aattacagct
     1141 cttaaggcta gagtattaat acgactcact atagggctag cgatcgccat ggagcccagc
     1201 agctggagcg gcagtgagag ccctgccgaa aacatggaaa ggatgagtga ctctgcagat
     1261 aagccaattg acaatgatgc agaaggggtc tggagccccg acatcgagca aagctttcag
     1321 gaggccctgg ctatctatcc accatgtggg aggaggaaaa tcatcttatc agacgaaggc
     1381 aaaatgtatg gtaggaatga attgatagcc agatacatca aactcaggac aggcaagacg
     1441 aggaccagaa aacaggtgtc tagtcacatt caggttcttg ccagaaggaa atctcgtgat
     1501 tttcattcca agctaaagga tcagactgca aaggataagg ccctgcagca catggcggcc
     1561 atgtcctcag cccagatcgt ctcggccact gccattcata acaagctggg gctgcctggg
     1621 attccacgcc cgaccttccc aggggcgccg gggttctggc cgggaatgat tcaaacaggg
     1681 cagccaggat cctcacaaga cgtcaagcct tttgtgcagc aggcctaccc catccagcca
     1741 gcggtcacag cccccattcc agggtttgag cctgcatcgg ccccagctcc ctcagtccct
     1801 gcctggcaag gtcgctccat tggcacaacc aagcttcgcc tggtggaatt ttcagctttt
     1861 ctcgagcagc agcgagaccc agactcgtac aacaaacacc tcttcgtgca cattgggcat
     1921 gccaaccatt cttacagtga cccattgctt gaatcagtgg acattcgtca gatttatgac
     1981 aaatttcctg aaaagaaagg tggcttaaag gaactgtttg gaaagggccc tcaaaatgcc
     2041 ttcttcctcg taaaattctg ggctgattta aactgcaata ttcaagatga tgctggggct
     2101 ttttatggtg taaccagtca gtacgagagt tctgaaaata tgacagtcac ctgttccacc
     2161 aaagtttgct cctttgggaa gcaagtagta gaaaaagtag agacggagta tgcaaggttt
     2221 gagaatggcc gatttgtata ccgaataaac cgctccccaa tgtgtgaata tatgatcaac
     2281 ttcatccaca agctcaaaca cttaccagag aaatatatga tgaacagtgt tttggaaaac
     2341 ttcacaattt tattggtggt aacaaacagg gatacacaag aaactctact ctgcatggcc
     2401 tgtgtgtttg aagtttcaaa tagtgaacac ggagcacaac atcatattta caggcttgta
     2461 aaggacgttt ctctcgagcc aaccactgag gatctgtact ttcagagcga taacgatgga
     2521 tccgaaatcg gtactggctt tccattcgac ccccattatg tggaagtcct gggcgagcgc
     2581 atgcactacg tcgatgttgg tccgcgcgat ggcacccctg tgctgttcct gcacggtaac
     2641 ccgacctcct cctacgtgtg gcgcaacatc atcccgcatg ttgcaccgac ccatcgctgc
     2701 attgctccag acctgatcgg tatgggcaaa tccgacaaac cagacctggg ttatttcttc
     2761 gacgaccacg tccgcttcat ggatgccttc atcgaagccc tgggtctgga agaggtcgtc
     2821 ctggtcattc acgactgggg ctccgctctg ggtttccact gggccaagcg caatccagag
     2881 cgcgtcaaag gtattgcatt tatggagttc atccgcccta tcccgacctg ggacgaatgg
     2941 ccagaatttg cccgcgagac cttccaggcc ttccgcacca ccgacgtcgg ccgcaagctg
     3001 atcatcgatc agaacgtttt tatcgagggt acgctgccga tgggtgtcgt ccgcccgctg
     3061 actgaagtcg agatggacca ttaccgcgag ccgttcctga atcctgttga ccgcgagcca
     3121 ctgtggcgct tcccaaacga gctgccaatc gccggtgagc cagcgaacat cgtcgcgctg
     3181 gtcgaagaat acatggactg gctgcaccag tcccctgtcc cgaagctgct gttctggggc
     3241 accccaggcg ttctgatccc accggccgaa gccgctcgcc tggccaaaag cctgcctaac
     3301 tgcaaggctg tggacatcgg cccgggtctg aatctgctgc aagaagacaa cccggacctg
     3361 atcggcagcg agatcgcgcg ctggctgtct actctggaga tttccggtta atagaattct
     3421 agagtcgacc tgcaggcatg caagctgatt tggctgctgc caccgctgag caaggccgct
     3481 tcgagcagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     3541 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     3601 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga
     3661 gatgtgggag gtttttttaa gcaagtaaaa cctctacaaa tgtggtaaaa tcttaattaa
     3721 gaattctaat ataacttcgt atagcataca ttatacgaag ttatggatcc actgttgaca
     3781 attaatcatc ggcatagtat atcggcatag tataatacga caaggtgagg aactaaaccc
     3841 aggaggcaga tcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     3901 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     3961 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     4021 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     4081 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     4141 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     4201 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     4261 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     4321 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     4381 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     4441 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     4501 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     4561 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     4621 tatcgccttc ttgacgagtt cttctgattc gaaatgaccg accaagcgac gcccaaccgg
     4681 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     4741 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     4801 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     4861 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     4921 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     4981 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     5041 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     5101 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     5161 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     5221 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     5281 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     5341 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggattt caagaagatc
     5401 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     5461 tggtcatgag attatcaaaa aggatcttca cctagatcct tttatagtcc ggaatcgtca
     5521 cacaaaaaac caacacacag atgtaatgaa aataaagata ttttattgcg gccatcgtga
     5581 tggctagagt ctaaactcga cctacctcct tagcgcgcgg tcgtgtactg ggccggcggt
     5641 tgccgctgtc ggccagcacc tcgacgcagc cgtcggtcca tacggctgcg ctccggcgag
     5701 cgatttgtgt gcgaccgacg gtgccggccc cgctgcggac gatggcatcg cagcggcctt
     5761 gagcccaggc agcatcgtcg aagttgccgt ccacgaggct ctggtagagc tgatccaggc
     5821 cgatgcgcag catgtaggct cgcagacgag ggctgccggc cagctcggga tgccggcgct
     5881 cgaagtagcg agtctgctgc tccatgcaag ccagccaggg ccgccagaag aagatgttgg
     5941 ccacctcgta ctgactgtcc ccgaacatag cttcggacca gtcgattacg gcggtgatgc
     6001 ggccgttgtc ggtcaggacg ttgttgctgc cgaagtcggc atggacgagg tggcgcacct
     6061 cgggacagtc ttcggcccac agcatcagtt cgtccagggc ttgagctacg ctggcggaca
     6121 cggtgtcgtc catcacggtc tgccagtggt agacatgggg atcagcaatg gcgcaaatga
     6181 aatcccgcca ggttgtgtac tggccgatgc cttggggccc gaatgggccg aagccgctgg
     6241 tttgactgag gtcggcggcg gcaatagcat ccatagcttc ggcgacaggc tgtaacacag
     6301 caggcagctc tgtttcgggg aggtcttgga gagtaacgcc ttgggcgcgt ctactgatgc
     6361 agtatgtcag gctctcgctg aactcgccga tgtccaacac ttcggggatg ggtagtgcag
     6421 cgctggcgaa gtggcggtac acgtagcggt ctttgtagaa gccatcagcg cagctattca
     6481 cccgcagtac atagccgcgt ccgccgacat cgaagctgaa ggctcggctc tcttcgccct
     6541 ccgacaactg catcaggtcg ctcacactgt cgaacttctc gatgagaaat ttttcaacgc
     6601 tggtagcggt gagttcgggc ttcttcatgg tggcatcgat tcccggtcaa catccagtac
     6661 acctccggca ccaccggctt ccccaagggc gccacgctct cccaccacaa catcctcaac
     6721 aacggctact gggtgggccg cacggtcggc tacacggagc aggaccgggt gtgtctgccg
     6781 gtgcccttct accactgctt cggcatggtc atgggcaact tgggcgccac ctcccacgcc
     6841 gcctgcatcg tcatcccggc cccgtccttc gagccggcgg cgacgctgga ggcggtgcag
     6901 cgggagaggt gcacgtccct gtacggcgtc cccacgaagc cgaattccac cacactggac
     6961 tagatgcatg ctcgacagtg tgatggatat ctgcagaatt cggctttcac gacacctgaa
     7021 atggaagaaa aaaactttga accactgtct gaggcttgag aatgaaccaa gatccaaact
     7081 caaaaagggc aaattccaag gagaattaca tcaagtgcca agctggccta acttcagtct
     7141 ccacccactc agtgtgggga aactccatcg cataaaaccc ctccccccaa cctaaagacg
     7201 acgtactcca aaagctcgag aactaatcga ggtgcctgga cggcgcccgg tactccgtgg
     7261 agtcacatga agcgacggct gaggacggaa aggccctttt cctttgtgtg ggtgactcac
     7321 ccgcccgctc tcccgagcgc cgcgtcctcc attttgagct gcctgcagca gggccgggaa
     7381 gcggccatct ttccgctcac gcaactggtg ccgaccgggc cagccttgcc gcccagggcg
     7441 gggcgataca cggcggcgcg aggccaggca ccagagcagg ccggccagct tgagactacc
     7501 cccgtccgat tctcggtggc cgcgctcgca ggccccgcct cgccgaacat gtgcgctggg
     7561 acgcacgggc cccgtcgccg cccgcggccc caaaaaccga aataccagtg tgcagatctt
     7621 ggcccgcatt tacaagacta tcttgccaga aaaaaagcgt cgcagcaggt catcaaaaat
     7681 tttaaatggc tagagactta tcgaaagcag cgagacaggc gcgaaggtgc caccagattc
     7741 gcacgcggcg gccccagcgc ccaggccagg cctcaactca agcacgaggc gaaggggctc
     7801 cttaagcgca aggcctcgaa ctctcccacc cacttccaac ccgaagctcg ggatcaagaa
     7861 tcacgtactg cagccaggtg gaagtaattc aaggcacgca agggccataa cccgtaaaga
     7921 ggccaggccc gcgggaacca cacacggcac ttacctgtgt tctggcggca aacccgttgc
     7981 gaaaaagaac gttcacggcg actactgcac ttatatacgg ttctccccca ccctcgggaa
     8041 aaaggcggag ccagtacacg acatcacttt cccagtttac cccgcgccac cttctctagg
     8101 caccggttca attgccgacc cctcccccca acttctcggg gactgtgggc gatgtgcgct
     8161 ctgcccactg acgggcaccg gagcc
//
