LOCUS       sequence-474682-        6423 bp    DNA     circular SYN 05-JUN-2025
DEFINITION  Express TEAD2-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 6423)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 6423)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 5, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: TEAD2-HaloTag Vector
FEATURES             Location/Qualifiers
     source          1..6423
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          644..776
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     759..778
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     820..839
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        820..838
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2214..2234
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             2250..3134
                     /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    3221..3342
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3232..3251)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(3457..3490)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3500..3547
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3579..4373
                     /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(3633..3652)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4243..4262
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4531..5119)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4611..4630)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5245..5293
                     /note="synthetic polyadenylation signal"
     CDS             complement(5317..5715)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5712..5721
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5746..5942)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        complement(5754..5964)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5760..5895
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5988..6008
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    6110..6191
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtccgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta cgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacaccaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta ataaccccgc cccgttgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatca ctagaagctt
      541 tattgcggta gtttatcaca gttaaattgc taacgcagtc agtgcttctg acacaacagt
      601 ctcgaactta agctgcagaa gttggtcgtg aggcactggg caggtaagta tcaaggttac
      661 aagacaggtt taaggagacc aatagaaact gggcttgtcg agacagagaa gactcttgcg
      721 tttctgatag gcacctattg gtcttactga catccacttt gcctttctct ccacaggtgt
      781 ccactcccag ttcaattaca gctcttaagg ctagagtatt aatacgactc actatagggc
      841 tagcgatcgc catgggggaa ccccgggctg gggccgccct ggacgatggc agcggctgga
      901 cgggcagtga ggaaggcagt gaggagggta ccggcggcag tgagggggct gggggtgacg
      961 ggggcccgga tgcagagggg gtgtggagcc cagacattga gcagagcttc caggaggccc
     1021 tggccatcta tccaccctgc ggccgccgga aaataatttt gtctgatgaa ggcaagatgt
     1081 atggtcggaa tgaactgatc gcccgctaca tcaagctgag aacggggaag acccgaactc
     1141 gaaaacaggt ttctagtcac atccaggttt tggcccgaag gaaatcaagg gaaatccagt
     1201 ccaagttgaa ggaccaggtt tccaaggaca aggctttcca gacaatggca accatgtcct
     1261 ctgcccagct catctccgcg ccttctctgc aggccaaact gggtcccact ggtcctcagg
     1321 cctctgagct tttccagttt tggtctggag gatctgggcc cccctggaat gttccagatg
     1381 tgaagccatt ctcacagaca ccgttcacct tgtcactgac tcccccatct actgacctcc
     1441 cagggtacga gcccccccaa gccctctcac ccctgccccc acctacccca tcgcccccag
     1501 cctggcaggc tcggggcctg ggcaccgccc ggttgcagct ggtagagttc tcagccttcg
     1561 tggaaccgcc agatgcagtt gattcttacc agaggcacct gttcgtgcac atcagccagc
     1621 actgccccag ccccggagcg ccgccgctcg agagtgtgga cgtccggcag atctacgaca
     1681 aattccctga gaaaaagggt ggcctccgag agctatatga tcgtggcccc ccccatgcct
     1741 tcttcctggt caagttctgg gcggacctga actggggccc aagtggtgag gaggcagggg
     1801 ccggtggcag catcagcagt ggtggcttct acggagtgag cagccagtat gagagcctgg
     1861 aacacatgac cctcacctgt tcctccaagg tctgctcttt tggcaagcag gtggtggaga
     1921 aggtggagac ggaacgggcc cagctggagg acggcagatt tgtgtaccgc ctgctgcgct
     1981 cgcccatgtg cgagtacctg gtgaatttct tgcacaagtt gcggcagctg cctgagcgat
     2041 acatgatgaa cagcgtcctg gaaaacttca ccatcctcca ggtggtgaca aacagagaca
     2101 cccaggaact gctgctctgc accgcctatg tcttcgaggt ctccaccagc gagcgtgggg
     2161 cccagcatca catttaccgc ctggtcaggg acgtttctct cgagccaacc actgaggatc
     2221 tgtactttca gagcgataac gatggatccg aaatcggtac tggctttcca ttcgaccccc
     2281 attatgtgga agtcctgggc gagcgcatgc actacgtcga tgttggtccg cgcgatggca
     2341 cccctgtgct gttcctgcac ggtaacccga cctcctccta cgtgtggcgc aacatcatcc
     2401 cgcatgttgc accgacccat cgctgcattg ctccagacct gatcggtatg ggcaaatccg
     2461 acaaaccaga cctgggttat ttcttcgacg accacgtccg cttcatggat gccttcatcg
     2521 aagccctggg tctggaagag gtcgtcctgg tcattcacga ctggggctcc gctctgggtt
     2581 tccactgggc caagcgcaat ccagagcgcg tcaaaggtat tgcatttatg gagttcatcc
     2641 gccctatccc gacctgggac gaatggccag aatttgcccg cgagaccttc caggccttcc
     2701 gcaccaccga cgtcggccgc aagctgatca tcgatcagaa cgtttttatc gagggtacgc
     2761 tgccgatggg tgtcgtccgc ccgctgactg aagtcgagat ggaccattac cgcgagccgt
     2821 tcctgaatcc tgttgaccgc gagccactgt ggcgcttccc aaacgagctg ccaatcgccg
     2881 gtgagccagc gaacatcgtc gcgctggtcg aagaatacat ggactggctg caccagtccc
     2941 ctgtcccgaa gctgctgttc tggggcaccc caggcgttct gatcccaccg gccgaagccg
     3001 ctcgcctggc caaaagcctg cctaactgca aggctgtgga catcggcccg ggtctgaatc
     3061 tgctgcaaga agacaacccg gacctgatcg gcagcgagat cgcgcgctgg ctgtctactc
     3121 tggagatttc cggttaatag aattctagag tcgacctgca ggcatgcaag ctgatttggc
     3181 tgctgccacc gctgagcaag gccgcttcga gcagacatga taagatacat tgatgagttt
     3241 ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct
     3301 attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt
     3361 cattttatgt ttcaggttca gggggagatg tgggaggttt ttttaagcaa gtaaaacctc
     3421 tacaaatgtg gtaaaatctt aattaagaat tctaatataa cttcgtatag catacattat
     3481 acgaagttat ggatccactg ttgacaatta atcatcggca tagtatatcg gcatagtata
     3541 atacgacaag gtgaggaact aaacccagga ggcagatcat gattgaacaa gatggattgc
     3601 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     3661 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     3721 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     3781 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     3841 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     3901 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     3961 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     4021 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     4081 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     4141 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     4201 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     4261 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     4321 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgattcgaaa
     4381 tgaccgacca agcgacgccc aaccggtatc agctcactca aaggcggtaa tacggttatc
     4441 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     4501 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     4561 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     4621 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     4681 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     4741 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     4801 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     4861 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     4921 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     4981 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     5041 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     5101 cagaaaaaaa ggatttcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     5161 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     5221 gatcctttta tagtccggaa tcgtcacaca aaaaaccaac acacagatgt aatgaaaata
     5281 aagatatttt attgcggcca tcgtgatggc tagagtttag ccctcccaca cataaccaga
     5341 gggcagcaat tcacgaatcc caactgccgt cggctgtcca tcactgtcct tcactatggc
     5401 tttgatccca ggatgcagat cgagaagcac ctgtcggcac cgtccgcagg ggctcaagat
     5461 gcccctgttc tcatttccga tcgcgacgat acaagtcagg ttgccagctg ccgcagcagc
     5521 agcagtgccc agcaccacga gttctgcaca aggtccccca gtaaaatgat atacattgac
     5581 accagtgaag atgcggccgt cgctagagag agctgcgctg gcgacgctgt agtcttcaga
     5641 gatggggatg ctgttgattg tagccgttgc tctttcaatg agggtggatt cttcttgaga
     5701 caaaggcttg gccatggtgg cgctagtgtc agaagaatcg agctttttgc aaaagcctag
     5761 gcctccaaaa aagcctcctc actacttctg gaatagctca gaggccgagg cggcctcggc
     5821 ctctgcataa ataaaaaaaa ttagtcagcc atggggcgga gaatgggcgg aactgggcgg
     5881 agttaggggc gggatgggcg gagttagggg cgggactatg gttgctgact aattgagatg
     5941 catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc ctaactgaca
     6001 cacattccac agctggttct ttccgcctca gaaggtacct aaccaagttc ctctttcaga
     6061 ggttatttca ggccatggtg ctgcgcaaga cgcgttaatt aagacgtcgg cagtgaaaaa
     6121 aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca
     6181 ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt
     6241 gggaggtttt ttaaagcaag taaaacctct acaaatgtgg tatggctgat tatgatccgc
     6301 ggccgcaata cgtcgacgtt atcagctgac ttcgtacgag agcctaggat tatggcgcgc
     6361 cactagttat taatagtaat caattacggg gtcattagtt catagcccat atatggagtt
     6421 ccg
//
