LOCUS       sequence-471870-        6630 bp    DNA     circular SYN 01-JUL-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 6630)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 6630)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 1, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: TEAD4-HaloTag BiBRET Vector
FEATURES             Location/Qualifiers
     source          1..6630
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    3..84
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     enhancer        317..620
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        621..824
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     775..795
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          960..1092
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1075..1094
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1136..1155
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1136..1154
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2491..2511
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             2527..3411
                     /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    3498..3619
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3509..3528)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(3734..3767)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3777..3824
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3856..4650
                     /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(3910..3929)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4520..4539
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      4867..5412
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     5313..5332
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5529..5546
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    5696..5744
                     /note="synthetic polyadenylation signal"
     CDS             complement(5768..6166)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      6163..6172
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(6197..6554)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6211..6346
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        6363..6554
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     6511..6531
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 cggcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
       61 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
      121 cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtatggct
      181 gattatgatc cgcggccgca atacgtcgac gttatcagct gacttcgtac gagagcctag
      241 gattatggcg cgccactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
      301 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      361 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      421 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      481 aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      541 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
      601 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
      661 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      721 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa
      781 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc
      841 agatcactag aagctttatt gcggtagttt atcacagtta aattgctaac gcagtcagtg
      901 cttctgacac aacagtctcg aacttaagct gcagaagttg gtcgtgaggc actgggcagg
      961 taagtatcaa ggttacaaga caggtttaag gagaccaata gaaactgggc ttgtcgagac
     1021 agagaagact cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct
     1081 ttctctccac aggtgtccac tcccagttca attacagctc ttaaggctag agtattaata
     1141 cgactcacta tagggctagc gatcgccatg gagggcacgg ccggcaccat tacctccaac
     1201 gagtggagct ctcccacctc ccctgagggg agcaccgcct ctgggggcag tcaggcactg
     1261 gacaagccca tcgacaatga cgcagagggc gtgtggagcc cggatattga gcagagtttc
     1321 caggaggccc tcgccatcta cccgccctgt ggcaggcgca aaatcatcct gtcggacgag
     1381 ggcaagatgt atggtcggaa cgagctgatt gcccgctaca tcaagctccg gacagggaag
     1441 acccgcacca ggaagcaggt ctccagccac atccaggtgc tggctcgtcg caaagctcgc
     1501 gagatccagg ccaagctaaa ggaccaggca gctaaggaca aggccctgca gagcatggct
     1561 gccatgtcgt ctgcacagat catctccgcc acggccttcc acagtagcat ggccctcgcc
     1621 cggggccccg gccgcccagc agtctcaggg ttttggcaag gagctttgcc aggccaagcc
     1681 ggaacgtccc atgatgtgaa gcctttctct cagcaaacct atgctgtcca gcctccgctg
     1741 cctctgccag ggtttgagtc tcctgcaggg cccgccccat cgccctctgc gcccccggca
     1801 cccccatggc agggccgcag cgtggccagc tccaagctct ggatgttgga gttctctgcc
     1861 ttcctggagc agcagcagga cccggacacg tacaacaagc acctgttcgt gcacattggc
     1921 cagtccagcc caagctacag cgacccctac ctcgaagccg tggacatccg ccaaatctat
     1981 gacaaattcc cggagaaaaa gggtggactc aaggatctct tcgaacgggg accctccaat
     2041 gccttttttc ttgtgaagtt ctgggcagac ctcaacacca acatcgagga tgaaggcagc
     2101 tccttctatg gggtctccag ccagtatgag agccccgaga acatgatcat cacctgctcc
     2161 acgaaggtct gctctttcgg caagcaggtg gtggagaaag ttgagacaga gtatgctcgc
     2221 tatgagaatg gacactactc ttaccgcatc caccggtccc cgctctgtga gtacatgatc
     2281 aacttcatcc acaagctcaa gcacctccct gagaagtaca tgatgaacag cgtgctggag
     2341 aacttcacca tcctgcaggt ggtcaccaac agagacacac aggagacctt gctgtgcatt
     2401 gcctatgtct ttgaggtgtc agccagtgag cacggggctc agcaccacat ctacaggctg
     2461 gtgaaagaag tttctctcga gccaaccact gaggatctgt actttcagag cgataacgat
     2521 ggatccgaaa tcggtactgg ctttccattc gacccccatt atgtggaagt cctgggcgag
     2581 cgcatgcact acgtcgatgt tggtccgcgc gatggcaccc ctgtgctgtt cctgcacggt
     2641 aacccgacct cctcctacgt gtggcgcaac atcatcccgc atgttgcacc gacccatcgc
     2701 tgcattgctc cagacctgat cggtatgggc aaatccgaca aaccagacct gggttatttc
     2761 ttcgacgacc acgtccgctt catggatgcc ttcatcgaag ccctgggtct ggaagaggtc
     2821 gtcctggtca ttcacgactg gggctccgct ctgggtttcc actgggccaa gcgcaatcca
     2881 gagcgcgtca aaggtattgc atttatggag ttcatccgcc ctatcccgac ctgggacgaa
     2941 tggccagaat ttgcccgcga gaccttccag gccttccgca ccaccgacgt cggccgcaag
     3001 ctgatcatcg atcagaacgt ttttatcgag ggtacgctgc cgatgggtgt cgtccgcccg
     3061 ctgactgaag tcgagatgga ccattaccgc gagccgttcc tgaatcctgt tgaccgcgag
     3121 ccactgtggc gcttcccaaa cgagctgcca atcgccggtg agccagcgaa catcgtcgcg
     3181 ctggtcgaag aatacatgga ctggctgcac cagtcccctg tcccgaagct gctgttctgg
     3241 ggcaccccag gcgttctgat cccaccggcc gaagccgctc gcctggccaa aagcctgcct
     3301 aactgcaagg ctgtggacat cggcccgggt ctgaatctgc tgcaagaaga caacccggac
     3361 ctgatcggca gcgagatcgc gcgctggctg tctactctgg agatttccgg ttaatagaat
     3421 tctagagtcg acctgcaggc atgcaagctg atttggctgc tgccaccgct gagcaaggcc
     3481 gcttcgagca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca
     3541 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
     3601 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg
     3661 ggagatgtgg gaggtttttt taagcaagta aaacctctac aaatgtggta aaatcttaat
     3721 taagaattct aatataactt cgtatagcat acattatacg aagttatgga tccactgttg
     3781 acaattaatc atcggcatag tatatcggca tagtataata cgacaaggtg aggaactaaa
     3841 cccaggaggc agatcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     3901 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     3961 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     4021 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     4081 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     4141 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     4201 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     4261 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     4321 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     4381 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     4441 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     4501 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     4561 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     4621 ttctatcgcc ttcttgacga gttcttctga ttcgaaatga ccgaccaagc gacgcccaac
     4681 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     4741 gaaagaacat gtcaaacatg agaattacaa cttatatcgt atggggctga cttcaggtgc
     4801 tacatttgaa gagataaatt gcactgaaat ctagaaatat tttatctgat taataagatg
     4861 atcttcttga gatcgttttg gtctgcgcgt aatctcttgc tctgaaaacg aaaaaaccgc
     4921 cttgcagggc ggtttttcga aggttctctg agctaccaac tctttgaacc gaggtaactg
     4981 gcttggagga gcgcagtcac caaaacttgt cctttcagtt tagccttaac cggcgcatga
     5041 cttcaagact aactcctcta aatcaattac cagtggctgc tgccagtggt gcttttgcat
     5101 gtctttccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcggactgaa
     5161 cggggggttc gtgcatacag tccagcttgg agcgaactgc ctacccggaa ctgagtgtca
     5221 ggcgtggaat gagacaaacg cggccataac agcggaatga caccggtaaa ccgaaaggca
     5281 ggaacaggag agcgcacgag ggagccgcca gggggaaacg cctggtatct ttatagtcct
     5341 gtcgggtttc gccaccactg atttgagcgt cagatttcgt gatgcttgtc aggggggcgg
     5401 agcctatgga aaaacggctt tgccgcggcc ctctcacttc cctgttaagt atcttcctgg
     5461 catcttccag gaaatctccg ccccgttcgt aagccatttc cgctcgccgc agtcgaacga
     5521 ccgagcgtag cgagtcagtg agcgaggaag cggaatatat cctgtatcac atattctgct
     5581 gacgcaccgg tgcagccttt tttctcctgc cacatgaagc acttcactga caccctcatc
     5641 agtgccaaca tagtaagcca gtatacactc cgctagcgct gatgtccgga atcgtcacac
     5701 aaaaaaccaa cacacagatg taatgaaaat aaagatattt tattgcggcc atcgtgatgg
     5761 ctagagttta gccctcccac acataaccag agggcagcaa ttcacgaatc ccaactgccg
     5821 tcggctgtcc atcactgtcc ttcactatgg ctttgatccc aggatgcaga tcgagaagca
     5881 cctgtcggca ccgtccgcag gggctcaaga tgcccctgtt ctcatttccg atcgcgacga
     5941 tacaagtcag gttgccagct gccgcagcag cagcagtgcc cagcaccacg agttctgcac
     6001 aaggtccccc agtaaaatga tatacattga caccagtgaa gatgcggccg tcgctagaga
     6061 gagctgcgct ggcgacgctg tagtcttcag agatggggat gctgttgatt gtagccgttg
     6121 ctctttcaat gagggtggat tcttcttgag acaaaggctt ggccatggtg gcgctagtgt
     6181 cagaagaatc gagctttttg caaaagccta ggcctccaaa aaagcctcct cactacttct
     6241 ggaatagctc agaggccgag gcggcctcgg cctctgcata aataaaaaaa attagtcagc
     6301 catggggcgg agaatgggcg gaactgggcg gagttagggg cgggatgggc ggagttaggg
     6361 gcgggactat ggttgctgac taattgagat gcatgctttg catacttctg cctgctgggg
     6421 agcctgggga ctttccacac ctggttgctg actaattgag atgcatgctt tgcatacttc
     6481 tgcctgctgg ggagcctggg gactttccac accctaactg acacacattc cacagctggt
     6541 tctttccgcc tcagaaggta cctaaccaag ttcctctttc agaggttatt tcaggccatg
     6601 gtgctgcgca agacgcgtta attaagacgt
//
