LOCUS       sequence-479889-        7246 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Express HaloTag-RAD21 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 7246)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7246)
  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            "Unpublished"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-RAD21 Vector
FEATURES             Location/Qualifiers
     source          1..7246
                     /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"
     regulatory      1164..1173
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1170..2060
                     /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             2073..2093
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     polyA_signal    4114..4235
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4125..4144)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(4350..4383)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        4393..4440
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4472..5266
                     /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(4526..4545)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5136..5155
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      5483..6028
                     /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     5929..5948
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6145..6162
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    6312..6360
                     /note="synthetic polyadenylation signal"
     CDS             complement(6384..6782)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      6779..6788
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(6813..7170)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6827..6962
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        6979..7170
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     7127..7147
                     /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 aaagccacca tggcagaaat cggtactggc tttccattcg
     1201 acccccatta tgtggaagtc ctgggcgagc gcatgcacta cgtcgatgtt ggtccgcgcg
     1261 atggcacccc tgtgctgttc ctgcacggta acccgacctc ctcctacgtg tggcgcaaca
     1321 tcatcccgca tgttgcaccg acccatcgct gcattgctcc agacctgatc ggtatgggca
     1381 aatccgacaa accagacctg ggttatttct tcgacgacca cgtccgcttc atggatgcct
     1441 tcatcgaagc cctgggtctg gaagaggtcg tcctggtcat tcacgactgg ggctccgctc
     1501 tgggtttcca ctgggccaag cgcaatccag agcgcgtcaa aggtattgca tttatggagt
     1561 tcatccgccc tatcccgacc tgggacgaat ggccagaatt tgcccgcgag accttccagg
     1621 ccttccgcac caccgacgtc ggccgcaagc tgatcatcga tcagaacgtt tttatcgagg
     1681 gtacgctgcc gatgggtgtc gtccgcccgc tgactgaagt cgagatggac cattaccgcg
     1741 agccgttcct gaatcctgtt gaccgcgagc cactgtggcg cttcccaaac gagctgccaa
     1801 tcgccggtga gccagcgaac atcgtcgcgc tggtcgaaga atacatggac tggctgcacc
     1861 agtcccctgt cccgaagctg ctgttctggg gcaccccagg cgttctgatc ccaccggccg
     1921 aagccgctcg cctggccaaa agcctgccta actgcaaggc tgtggacatc ggcccgggtc
     1981 tgaatctgct gcaagaagac aacccggacc tgatcggcag cgagatcgcg cgctggctgt
     2041 cgacgctcga gatttccggc gagccaacca ctgaggatct gtactttcag agcgataacg
     2101 cgatcgccat gttctacgca cattttgttc tcagtaaaag agggcctctg gccaaaattt
     2161 ggctagcggc ccattgggat aagaagctaa ccaaagccca tgtgttcgag tgtaatttag
     2221 agagcagcgt ggagagtatc atctcaccaa aggtgaaaat ggcattacgg acatcaggac
     2281 atctcttact gggagtagtt cgaatctatc acaggaaagc caaatacctt cttgcagact
     2341 gtaatgaagc attcattaag ataaagatgg cttttcggcc aggtgtggtt gacctgcctg
     2401 aggaaaatcg ggaagcagct tataatgcca ttactttacc tgaagaattt catgactttg
     2461 atcagccact gcctgactta gatgacatcg atgtggccca gcagttcagc ttgaatcaga
     2521 gtagagtgga agagataacc atgagagaag aagttgggaa catcagtatt ttacaagaaa
     2581 atgattttgg tgattttgga atggatgatc gtgagataat gagagaaggc agtgcttttg
     2641 aggatgacga catgttagta agcactacta cttctaacct cctattagag tctgaacaga
     2701 gcaccagcaa tctgaatgag aaaattaacc atttagaata tgaagatcaa tataaggatg
     2761 ataattttgg agaaggaaat gatggtggaa tattagatga caaacttatt agtaataatg
     2821 atggcggtat ctttgatgat ccccctgccc tctctgaggc aggggtgatg ttgccagagc
     2881 agcctgcaca tgacgatatg gatgaggatg ataatgtatc aatgggtggg cctgatagtc
     2941 ctgattcagt ggatcccgtt gaaccaatgc caaccatgac tgatcaaaca acacttgttc
     3001 caaatgagga agaagcattt gcattggagc ctattgatat aactgttaaa gaaacaaaag
     3061 ccaagaggaa gaggaagcta attgttgaca gtgtcaaaga gttggatagc aagacaatta
     3121 gagcccaact tagtgattat tcagatattg ttactacttt ggatctggca ccgcccacca
     3181 agaaattgat gatgtggaaa gagacaggag gagtagaaaa actgttttct ttacctgctc
     3241 agcctttgtg gaataacaga ctactgaagc tctttacacg ctgtcttaca ccgcttgtac
     3301 cagaagacct tagaaaaagg aggaaaggag gagaggcaga taatttggat gaattcctca
     3361 aagaatttga aaatccagag gttcctagag aggaccagca acagcagcat cagcagcgtg
     3421 atgttatcga tgagcccatt attgaagagc caagccgcct ccaggagtca gtgatggagg
     3481 ccagcagaac aaacatagat gagtcagcta tgcctccacc accacctcag ggagttaagc
     3541 gaaaagctgg acaaattgac ccagagcctg tgatgcctcc tcagcaggta gagcagatgg
     3601 aaataccacc tgtagagctt cccccagaag aacctccaaa tatctgtcag ctaataccag
     3661 agttagaact tctgccagaa aaagagaagg agaaagagaa ggaaaaagaa gatgatgaag
     3721 aggaagagga tgaagatgca tcagggggcg atcaagatca ggaagaaaga agatggaaca
     3781 aaaggactca gcagatgctt catggtcttc agcgtgctct tgctaaaact ggagctgaat
     3841 ctatcagttt gcttgagtta tgtcgaaata cgaacagaaa acaagctgcc gcaaagttct
     3901 acagcttctt ggttcttaaa aagcagcaag ctattgagct gacacaggaa gaaccgtaca
     3961 gtgacatcat cgcaacacct ggaccaaggt tccatattat agtttaaacg aattcgggct
     4021 cggtacccgg ggatcctcta gagtcgacct gcaggcatgc aagctgattt ggctgctgcc
     4081 accgctgagc aaggccgctt cgagcagaca tgataagata cattgatgag tttggacaaa
     4141 ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt
     4201 tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta
     4261 tgtttcaggt tcagggggag atgtgggagg tttttttaag caagtaaaac ctctacaaat
     4321 gtggtaaaat cttaattaag aattctaata taacttcgta tagcatacat tatacgaagt
     4381 tatggatcca ctgttgacaa ttaatcatcg gcatagtata tcggcatagt ataatacgac
     4441 aaggtgagga actaaaccca ggaggcagat catgattgaa caagatggat tgcacgcagg
     4501 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     4561 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     4621 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct
     4681 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     4741 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     4801 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     4861 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     4921 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     4981 gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     5041 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     5101 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     5161 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     5221 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgattcg aaatgaccga
     5281 ccaagcgacg cccaaccggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     5341 tcaggggata acgcaggaaa gaacatgtca aacatgagaa ttacaactta tatcgtatgg
     5401 ggctgacttc aggtgctaca tttgaagaga taaattgcac tgaaatctag aaatatttta
     5461 tctgattaat aagatgatct tcttgagatc gttttggtct gcgcgtaatc tcttgctctg
     5521 aaaacgaaaa aaccgccttg cagggcggtt tttcgaaggt tctctgagct accaactctt
     5581 tgaaccgagg taactggctt ggaggagcgc agtcaccaaa acttgtcctt tcagtttagc
     5641 cttaaccggc gcatgacttc aagactaact cctctaaatc aattaccagt ggctgctgcc
     5701 agtggtgctt ttgcatgtct ttccgggttg gactcaagac gatagttacc ggataaggcg
     5761 cagcggtcgg actgaacggg gggttcgtgc atacagtcca gcttggagcg aactgcctac
     5821 ccggaactga gtgtcaggcg tggaatgaga caaacgcggc cataacagcg gaatgacacc
     5881 ggtaaaccga aaggcaggaa caggagagcg cacgagggag ccgccagggg gaaacgcctg
     5941 gtatctttat agtcctgtcg ggtttcgcca ccactgattt gagcgtcaga tttcgtgatg
     6001 cttgtcaggg gggcggagcc tatggaaaaa cggctttgcc gcggccctct cacttccctg
     6061 ttaagtatct tcctggcatc ttccaggaaa tctccgcccc gttcgtaagc catttccgct
     6121 cgccgcagtc gaacgaccga gcgtagcgag tcagtgagcg aggaagcgga atatatcctg
     6181 tatcacatat tctgctgacg caccggtgca gccttttttc tcctgccaca tgaagcactt
     6241 cactgacacc ctcatcagtg ccaacatagt aagccagtat acactccgct agcgctgatg
     6301 tccggaatcg tcacacaaaa aaccaacaca cagatgtaat gaaaataaag atattttatt
     6361 gcggccatcg tgatggctag agtttagccc tcccacacat aaccagaggg cagcaattca
     6421 cgaatcccaa ctgccgtcgg ctgtccatca ctgtccttca ctatggcttt gatcccagga
     6481 tgcagatcga gaagcacctg tcggcaccgt ccgcaggggc tcaagatgcc cctgttctca
     6541 tttccgatcg cgacgataca agtcaggttg ccagctgccg cagcagcagc agtgcccagc
     6601 accacgagtt ctgcacaagg tcccccagta aaatgatata cattgacacc agtgaagatg
     6661 cggccgtcgc tagagagagc tgcgctggcg acgctgtagt cttcagagat ggggatgctg
     6721 ttgattgtag ccgttgctct ttcaatgagg gtggattctt cttgagacaa aggcttggcc
     6781 atggtggcgc tagtgtcaga agaatcgagc tttttgcaaa agcctaggcc tccaaaaaag
     6841 cctcctcact acttctggaa tagctcagag gccgaggcgg cctcggcctc tgcataaata
     6901 aaaaaaatta gtcagccatg gggcggagaa tgggcggaac tgggcggagt taggggcggg
     6961 atgggcggag ttaggggcgg gactatggtt gctgactaat tgagatgcat gctttgcata
     7021 cttctgcctg ctggggagcc tggggacttt ccacacctgg ttgctgacta attgagatgc
     7081 atgctttgca tacttctgcc tgctggggag cctggggact ttccacaccc taactgacac
     7141 acattccaca gctggttctt tccgcctcag aaggtaccta accaagttcc tctttcagag
     7201 gttatttcag gccatggtgc tgcgcaagac gcgttaatta agacgt
//
