LOCUS       sequence-475150-        9102 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express STAG1-HaloTag 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 9102)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9102)
  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            "Unpublished"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: STAG1-HaloTag Vector
FEATURES             Location/Qualifiers
     source          1..9102
                     /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             4963..4983
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             4999..5883
                     /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    5970..6091
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5981..6000)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(6206..6239)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        6249..6296
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             6328..7122
                     /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(6382..6401)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6992..7011
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      7339..7884
                     /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     7785..7804
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8001..8018
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    8168..8216
                     /note="synthetic polyadenylation signal"
     CDS             complement(8240..8638)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      8635..8644
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(8669..9026)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8683..8818
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        8835..9026
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     8983..9003
                     /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 attacttcag aattaccagt gttacaggat
     1201 tcaactaatg aaactactgc ccattccgat gctggcagcg agcttgaaga aacagaggtc
     1261 aaaggaaaaa gaaaaagggg tcgtcctggc cggcctccat ctacaaataa gaaacctcga
     1321 aaatctccag gtgagaagag cagaattgaa gctggaatta gaggagcagg ccgtggaaga
     1381 gctaatggac accctcaaca gaatggggaa ggggagcctg tcacattatt tgaggtggtg
     1441 aaactgggga aaagtgcaat gcagtccgtg gtggatgact ggattgaatc atataaacaa
     1501 gacagggaca tcgcacttct ggatttaatc aactttttta tccagtgttc aggatgtcga
     1561 ggtactgtga gaatagagat gtttcgaaat atgcagaatg cagaaatcat cagaaaaatg
     1621 actgaagaat ttgatgagga cagtggtgat tatcctctta ccatgcctgg acctcagtgg
     1681 aaaaaatttc gttcaaactt ttgtgaattt attggagtcc tgattcgaca gtgtcagtat
     1741 agcataattt atgatgagta tatgatggac acagtaatct cccttttgac gggtttgtca
     1801 gactcccagg tcagagcttt taggcataca agtaccctgg ctgccatgaa gctcatgact
     1861 gctctggtga atgttgcctt aaacctcagt attcatcagg ataataccca gagacaatat
     1921 gaagccgaga gaaataaaat gattgggaag agagccaatg aaaggttgga gttactactt
     1981 cagaaacgca aagagctgca agaaaatcag gatgaaatcg aaaatatgat gaactctatt
     2041 tttaagggta tatttgttca tagataccgt gatgctattg ctgagattag agccatttgt
     2101 attgaagaaa ttggagtatg gatgaaaatg tatagtgatg ccttcctaaa tgacagttac
     2161 ctaaaatatg ttggctggac tcttcatgac aggcaagggg aagtcaggct gaagtgtttg
     2221 aaagctctgc agagtctata taccaataga gaattattcc ccaaattgga actattcact
     2281 aaccgattca aggatcgcat tgtatcaatg acacttgata aagaatatga tgttgctgtg
     2341 gaagctattc gattggttac tctgatactt catggaagtg aagaagctct ttccaatgaa
     2401 gactgtgaaa atgtttacca cttggtgtac tcggcacatc gccctgttgc tgtggcagct
     2461 ggagagttcc ttcacaaaaa gctatttagc agacatgacc cacaagcaga agaagcatta
     2521 gcaaagagga ggggaagaaa cagcccgaat ggaaacctca ttaggatgct ggttcttttc
     2581 tttcttgaaa gtgagttaca tgaacatgca gcctacttgg tggacagttt atgggagagc
     2641 tctcaagaac tgttgaaaga ctgggaatgt atgacagagt tgctattaga agaacctgtt
     2701 caaggagagg aagcaatgtc tgatcgtcaa gagagtgctc ttatagagct aatggtttgt
     2761 acaattcgtc aagctgctga ggcacatcct ccagtgggaa ggggtaccgg caagagagtg
     2821 ctaactgcca aagaaaggaa aactcaaatt gatgatagaa acaaattgac tgaacatttt
     2881 attattacac ttcctatgtt actgtcaaag tattctgcag atgcagagaa ggtagcaaac
     2941 ttgctacaaa tcccacagta ttttgattta gaaatctaca gcacaggtag aatggaaaag
     3001 catctggatg ctttattaaa acagattaag tttgttgtgg agaaacacgt agaatcagat
     3061 gttctagaag cctgcagtaa aacctatagt atcttatgca gtgaagaata taccatccag
     3121 aacagagttg acatagctcg aagccagctg attgatgagt ttgtagatcg attcaatcat
     3181 tctgtggaag acctattgca agagggagaa gaagctgatg atgatgacat ttacaatgtt
     3241 ctttctacat taaagcggtt aacttctttt cacaatgcac atgatctcac aaaatgggat
     3301 ctctttggta attgctacag attattgaag actggaattg aacatggagc catgccagaa
     3361 cagatagtcg tgcaagcact gcagtgttcc cattattcga ttctttggca gttggtgaaa
     3421 attactgatg gctctccttc caaagaggat ttgttggtat tgaggaaaac ggtgaaatcc
     3481 tttttggctg tttgccagca gtgcctgtct aatgttaata ctccagtgaa agaacaggct
     3541 ttcatgttac tctgtgatct tctgatgatt ttcagccacc aattaatgac aggtggcaga
     3601 gagggccttc agcctttggt gttcaatcca gatactggac tccaatctga actcctcagt
     3661 tttgtgatgg atcacgtttt tattgaccaa gacgaggaga accagagcat ggagggtgat
     3721 gaagaagatg aagctaataa aattgaggcc ttacataaaa gaaggaatct acttgctgct
     3781 ttcagcaaac ttatcattta tgacattgtt gacatgcatg cagctgcaga catcttcaaa
     3841 cactacatga agtattacaa tgactatggt gatattatta aggaaacact gagtaaaacc
     3901 aggcagattg ataaaattca gtgtgccaag actctcattc tcagtttgca acagttattt
     3961 aatgaacttg ttcaagagca aggtcccaac ctagatagga catctgccca tgtcagtggc
     4021 attaaagaac tggcacgtcg ctttgccctt acatttggat tggaccagat taagacacga
     4081 gaagcagttg ccacacttca caaggatggc atagagtttg catttaaata ccaaaatcag
     4141 aaaggacaag agtatccacc tcctaatctg gcttttcttg aagtactaag tgaattttct
     4201 tctaaacttc ttcgacagga caaaaagaca gttcattcat acctagagaa attccttacc
     4261 gagcagatga tggaaaggag ggaggatgta tggcttccac tcatctccta tagaaattca
     4321 ttagtcactg ggggtgaaga tgatagaatg tctgtgaaca gtggaagtag cagcagcaaa
     4381 acctcatcag taaggaataa gaaaggacga cctccacttc ataaaaaacg agtagaagat
     4441 gagagtctgg ataacacatg gctaaacagg actgacacca tgattcagac tcctggcccc
     4501 ctgccagcac cacaactcac atccactgta ctgcgggaga acagtcggcc catgggagac
     4561 cagattcaag aacctgagtc tgaacatggt tctgaaccag actttttaca caatcctcag
     4621 atgcagatct cttggttagg ccagccgaag ttagaagact taaatcggaa ggacagaaca
     4681 ggaatgaact acatgaaagt gagaactgga gtgaggcatg ctgttcgggg tctaatggag
     4741 gaagatgctg agcccatctt tgaagatgtg atgatgtcat cccgaagcca gttagaagat
     4801 atgaatgaag aatttgagga caccatggtt attgatctgc ctccatcaag aaatcggcga
     4861 gagagagctg agctaaggcc agacttcttt gactctgcag ctatcataga agatgattca
     4921 ggatttggaa tgcctatgtt cgtttctctc gagccaacca ctgaggatct gtactttcag
     4981 agcgataacg atggatccga aatcggtact ggctttccat tcgaccccca ttatgtggaa
     5041 gtcctgggcg agcgcatgca ctacgtcgat gttggtccgc gcgatggcac ccctgtgctg
     5101 ttcctgcacg gtaacccgac ctcctcctac gtgtggcgca acatcatccc gcatgttgca
     5161 ccgacccatc gctgcattgc tccagacctg atcggtatgg gcaaatccga caaaccagac
     5221 ctgggttatt tcttcgacga ccacgtccgc ttcatggatg ccttcatcga agccctgggt
     5281 ctggaagagg tcgtcctggt cattcacgac tggggctccg ctctgggttt ccactgggcc
     5341 aagcgcaatc cagagcgcgt caaaggtatt gcatttatgg agttcatccg ccctatcccg
     5401 acctgggacg aatggccaga atttgcccgc gagaccttcc aggccttccg caccaccgac
     5461 gtcggccgca agctgatcat cgatcagaac gtttttatcg agggtacgct gccgatgggt
     5521 gtcgtccgcc cgctgactga agtcgagatg gaccattacc gcgagccgtt cctgaatcct
     5581 gttgaccgcg agccactgtg gcgcttccca aacgagctgc caatcgccgg tgagccagcg
     5641 aacatcgtcg cgctggtcga agaatacatg gactggctgc accagtcccc tgtcccgaag
     5701 ctgctgttct ggggcacccc aggcgttctg atcccaccgg ccgaagccgc tcgcctggcc
     5761 aaaagcctgc ctaactgcaa ggctgtggac atcggcccgg gtctgaatct gctgcaagaa
     5821 gacaacccgg acctgatcgg cagcgagatc gcgcgctggc tgtctactct ggagatttcc
     5881 ggttaataga attctagagt cgacctgcag gcatgcaagc tgatttggct gctgccaccg
     5941 ctgagcaagg ccgcttcgag cagacatgat aagatacatt gatgagtttg gacaaaccac
     6001 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     6061 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
     6121 tcaggttcag ggggagatgt gggaggtttt tttaagcaag taaaacctct acaaatgtgg
     6181 taaaatctta attaagaatt ctaatataac ttcgtatagc atacattata cgaagttatg
     6241 gatccactgt tgacaattaa tcatcggcat agtatatcgg catagtataa tacgacaagg
     6301 tgaggaacta aacccaggag gcagatcatg attgaacaag atggattgca cgcaggttct
     6361 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     6421 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     6481 gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc
     6541 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     6601 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     6661 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     6721 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     6781 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     6841 gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     6901 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     6961 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     7021 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     7081 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gattcgaaat gaccgaccaa
     7141 gcgacgccca accggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     7201 gggataacgc aggaaagaac atgtcaaaca tgagaattac aacttatatc gtatggggct
     7261 gacttcaggt gctacatttg aagagataaa ttgcactgaa atctagaaat attttatctg
     7321 attaataaga tgatcttctt gagatcgttt tggtctgcgc gtaatctctt gctctgaaaa
     7381 cgaaaaaacc gccttgcagg gcggtttttc gaaggttctc tgagctacca actctttgaa
     7441 ccgaggtaac tggcttggag gagcgcagtc accaaaactt gtcctttcag tttagcctta
     7501 accggcgcat gacttcaaga ctaactcctc taaatcaatt accagtggct gctgccagtg
     7561 gtgcttttgc atgtctttcc gggttggact caagacgata gttaccggat aaggcgcagc
     7621 ggtcggactg aacggggggt tcgtgcatac agtccagctt ggagcgaact gcctacccgg
     7681 aactgagtgt caggcgtgga atgagacaaa cgcggccata acagcggaat gacaccggta
     7741 aaccgaaagg caggaacagg agagcgcacg agggagccgc cagggggaaa cgcctggtat
     7801 ctttatagtc ctgtcgggtt tcgccaccac tgatttgagc gtcagatttc gtgatgcttg
     7861 tcaggggggc ggagcctatg gaaaaacggc tttgccgcgg ccctctcact tccctgttaa
     7921 gtatcttcct ggcatcttcc aggaaatctc cgccccgttc gtaagccatt tccgctcgcc
     7981 gcagtcgaac gaccgagcgt agcgagtcag tgagcgagga agcggaatat atcctgtatc
     8041 acatattctg ctgacgcacc ggtgcagcct tttttctcct gccacatgaa gcacttcact
     8101 gacaccctca tcagtgccaa catagtaagc cagtatacac tccgctagcg ctgatgtccg
     8161 gaatcgtcac acaaaaaacc aacacacaga tgtaatgaaa ataaagatat tttattgcgg
     8221 ccatcgtgat ggctagagtt tagccctccc acacataacc agagggcagc aattcacgaa
     8281 tcccaactgc cgtcggctgt ccatcactgt ccttcactat ggctttgatc ccaggatgca
     8341 gatcgagaag cacctgtcgg caccgtccgc aggggctcaa gatgcccctg ttctcatttc
     8401 cgatcgcgac gatacaagtc aggttgccag ctgccgcagc agcagcagtg cccagcacca
     8461 cgagttctgc acaaggtccc ccagtaaaat gatatacatt gacaccagtg aagatgcggc
     8521 cgtcgctaga gagagctgcg ctggcgacgc tgtagtcttc agagatgggg atgctgttga
     8581 ttgtagccgt tgctctttca atgagggtgg attcttcttg agacaaaggc ttggccatgg
     8641 tggcgctagt gtcagaagaa tcgagctttt tgcaaaagcc taggcctcca aaaaagcctc
     8701 ctcactactt ctggaatagc tcagaggccg aggcggcctc ggcctctgca taaataaaaa
     8761 aaattagtca gccatggggc ggagaatggg cggaactggg cggagttagg ggcgggatgg
     8821 gcggagttag gggcgggact atggttgctg actaattgag atgcatgctt tgcatacttc
     8881 tgcctgctgg ggagcctggg gactttccac acctggttgc tgactaattg agatgcatgc
     8941 tttgcatact tctgcctgct ggggagcctg gggactttcc acaccctaac tgacacacat
     9001 tccacagctg gttctttccg cctcagaagg tacctaacca agttcctctt tcagaggtta
     9061 tttcaggcca tggtgctgcg caagacgcgt taattaagac gt
//
