LOCUS       sequence-479895-        9127 bp    DNA     circular SYN 01-AUG-2025
DEFINITION  Express HaloTag-STAG1 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 9127)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9127)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 1, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: HaloTag-STAG1 Vector
FEATURES             Location/Qualifiers
     source          1..9127
                     /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    5995..6116
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6006..6025)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(6231..6264)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        6274..6321
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             6353..7147
                     /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(6407..6426)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7017..7036
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      7364..7909
                     /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     7810..7829
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8026..8043
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    8193..8241
                     /note="synthetic polyadenylation signal"
     CDS             complement(8265..8663)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      8660..8669
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(8694..9051)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8708..8843
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        8860..9051
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     9008..9028
                     /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 gattacttca gaattaccag tgttacagga ttcaactaat gaaactactg
     2161 cccattccga tgctggcagc gagcttgaag aaacagaggt caaaggaaaa agaaaaaggg
     2221 gtcgtcctgg ccggcctcca tctacaaata agaaacctcg aaaatctcca ggtgagaaga
     2281 gcagaattga agctggaatt agaggagcag gccgtggaag agctaatgga caccctcaac
     2341 agaatgggga aggggagcct gtcacattat ttgaggtggt gaaactgggg aaaagtgcaa
     2401 tgcagtccgt ggtggatgac tggattgaat catataaaca agacagggac atcgcacttc
     2461 tggatttaat caactttttt atccagtgtt caggatgtcg aggtactgtg agaatagaga
     2521 tgtttcgaaa tatgcagaat gcagaaatca tcagaaaaat gactgaagaa tttgatgagg
     2581 acagtggtga ttatcctctt accatgcctg gacctcagtg gaaaaaattt cgttcaaact
     2641 tttgtgaatt tattggagtc ctgattcgac agtgtcagta tagcataatt tatgatgagt
     2701 atatgatgga cacagtaatc tcccttttga cgggtttgtc agactcccag gtcagagctt
     2761 ttaggcatac aagtaccctg gctgccatga agctcatgac tgctctggtg aatgttgcct
     2821 taaacctcag tattcatcag gataataccc agagacaata tgaagccgag agaaataaaa
     2881 tgattgggaa gagagccaat gaaaggttgg agttactact tcagaaacgc aaagagctgc
     2941 aagaaaatca ggatgaaatc gaaaatatga tgaactctat ttttaagggt atatttgttc
     3001 atagataccg tgatgctatt gctgagatta gagccatttg tattgaagaa attggagtat
     3061 ggatgaaaat gtatagtgat gccttcctaa atgacagtta cctaaaatat gttggctgga
     3121 ctcttcatga caggcaaggg gaagtcaggc tgaagtgttt gaaagctctg cagagtctat
     3181 ataccaatag agaattattc cccaaattgg aactattcac taaccgattc aaggatcgca
     3241 ttgtatcaat gacacttgat aaagaatatg atgttgctgt ggaagctatt cgattggtta
     3301 ctctgatact tcatggaagt gaagaagctc tttccaatga agactgtgaa aatgtttacc
     3361 acttggtgta ctcggcacat cgccctgttg ctgtggcagc tggagagttc cttcacaaaa
     3421 agctatttag cagacatgac ccacaagcag aagaagcatt agcaaagagg aggggaagaa
     3481 acagcccgaa tggaaacctc attaggatgc tggttctttt ctttcttgaa agtgagttac
     3541 atgaacatgc agcctacttg gtggacagtt tatgggagag ctctcaagaa ctgttgaaag
     3601 actgggaatg tatgacagag ttgctattag aagaacctgt tcaaggagag gaagcaatgt
     3661 ctgatcgtca agagagtgct cttatagagc taatggtttg tacaattcgt caagctgctg
     3721 aggcacatcc tccagtggga aggggtaccg gcaagagagt gctaactgcc aaagaaagga
     3781 aaactcaaat tgatgataga aacaaattga ctgaacattt tattattaca cttcctatgt
     3841 tactgtcaaa gtattctgca gatgcagaga aggtagcaaa cttgctacaa atcccacagt
     3901 attttgattt agaaatctac agcacaggta gaatggaaaa gcatctggat gctttattaa
     3961 aacagattaa gtttgttgtg gagaaacacg tagaatcaga tgttctagaa gcctgcagta
     4021 aaacctatag tatcttatgc agtgaagaat ataccatcca gaacagagtt gacatagctc
     4081 gaagccagct gattgatgag tttgtagatc gattcaatca ttctgtggaa gacctattgc
     4141 aagagggaga agaagctgat gatgatgaca tttacaatgt tctttctaca ttaaagcggt
     4201 taacttcttt tcacaatgca catgatctca caaaatggga tctctttggt aattgctaca
     4261 gattattgaa gactggaatt gaacatggag ccatgccaga acagatagtc gtgcaagcac
     4321 tgcagtgttc ccattattcg attctttggc agttggtgaa aattactgat ggctctcctt
     4381 ccaaagagga tttgttggta ttgaggaaaa cggtgaaatc ctttttggct gtttgccagc
     4441 agtgcctgtc taatgttaat actccagtga aagaacaggc tttcatgtta ctctgtgatc
     4501 ttctgatgat tttcagccac caattaatga caggtggcag agagggcctt cagcctttgg
     4561 tgttcaatcc agatactgga ctccaatctg aactcctcag ttttgtgatg gatcacgttt
     4621 ttattgacca agacgaggag aaccagagca tggagggtga tgaagaagat gaagctaata
     4681 aaattgaggc cttacataaa agaaggaatc tacttgctgc tttcagcaaa cttatcattt
     4741 atgacattgt tgacatgcat gcagctgcag acatcttcaa acactacatg aagtattaca
     4801 atgactatgg tgatattatt aaggaaacac tgagtaaaac caggcagatt gataaaattc
     4861 agtgtgccaa gactctcatt ctcagtttgc aacagttatt taatgaactt gttcaagagc
     4921 aaggtcccaa cctagatagg acatctgccc atgtcagtgg cattaaagaa ctggcacgtc
     4981 gctttgccct tacatttgga ttggaccaga ttaagacacg agaagcagtt gccacacttc
     5041 acaaggatgg catagagttt gcatttaaat accaaaatca gaaaggacaa gagtatccac
     5101 ctcctaatct ggcttttctt gaagtactaa gtgaattttc ttctaaactt cttcgacagg
     5161 acaaaaagac agttcattca tacctagaga aattccttac cgagcagatg atggaaagga
     5221 gggaggatgt atggcttcca ctcatctcct atagaaattc attagtcact gggggtgaag
     5281 atgatagaat gtctgtgaac agtggaagta gcagcagcaa aacctcatca gtaaggaata
     5341 agaaaggacg acctccactt cataaaaaac gagtagaaga tgagagtctg gataacacat
     5401 ggctaaacag gactgacacc atgattcaga ctcctggccc cctgccagca ccacaactca
     5461 catccactgt actgcgggag aacagtcggc ccatgggaga ccagattcaa gaacctgagt
     5521 ctgaacatgg ttctgaacca gactttttac acaatcctca gatgcagatc tcttggttag
     5581 gccagccgaa gttagaagac ttaaatcgga aggacagaac aggaatgaac tacatgaaag
     5641 tgagaactgg agtgaggcat gctgttcggg gtctaatgga ggaagatgct gagcccatct
     5701 ttgaagatgt gatgatgtca tcccgaagcc agttagaaga tatgaatgaa gaatttgagg
     5761 acaccatggt tattgatctg cctccatcaa gaaatcggcg agagagagct gagctaaggc
     5821 cagacttctt tgactctgca gctatcatag aagatgattc aggatttgga atgcctatgt
     5881 tcgtttaaac gaattcgggc tcggtacccg gggatcctct agagtcgacc tgcaggcatg
     5941 caagctgatt tggctgctgc caccgctgag caaggccgct tcgagcagac atgataagat
     6001 acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg
     6061 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca
     6121 acaacaattg cattcatttt atgtttcagg ttcaggggga gatgtgggag gtttttttaa
     6181 gcaagtaaaa cctctacaaa tgtggtaaaa tcttaattaa gaattctaat ataacttcgt
     6241 atagcataca ttatacgaag ttatggatcc actgttgaca attaatcatc ggcatagtat
     6301 atcggcatag tataatacga caaggtgagg aactaaaccc aggaggcaga tcatgattga
     6361 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     6421 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     6481 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga
     6541 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     6601 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     6661 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     6721 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     6781 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     6841 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga
     6901 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     6961 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     7021 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     7081 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     7141 cttctgattc gaaatgaccg accaagcgac gcccaaccgg tatcagctca ctcaaaggcg
     7201 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtc aaacatgaga
     7261 attacaactt atatcgtatg gggctgactt caggtgctac atttgaagag ataaattgca
     7321 ctgaaatcta gaaatatttt atctgattaa taagatgatc ttcttgagat cgttttggtc
     7381 tgcgcgtaat ctcttgctct gaaaacgaaa aaaccgcctt gcagggcggt ttttcgaagg
     7441 ttctctgagc taccaactct ttgaaccgag gtaactggct tggaggagcg cagtcaccaa
     7501 aacttgtcct ttcagtttag ccttaaccgg cgcatgactt caagactaac tcctctaaat
     7561 caattaccag tggctgctgc cagtggtgct tttgcatgtc tttccgggtt ggactcaaga
     7621 cgatagttac cggataaggc gcagcggtcg gactgaacgg ggggttcgtg catacagtcc
     7681 agcttggagc gaactgccta cccggaactg agtgtcaggc gtggaatgag acaaacgcgg
     7741 ccataacagc ggaatgacac cggtaaaccg aaaggcagga acaggagagc gcacgaggga
     7801 gccgccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc accactgatt
     7861 tgagcgtcag atttcgtgat gcttgtcagg ggggcggagc ctatggaaaa acggctttgc
     7921 cgcggccctc tcacttccct gttaagtatc ttcctggcat cttccaggaa atctccgccc
     7981 cgttcgtaag ccatttccgc tcgccgcagt cgaacgaccg agcgtagcga gtcagtgagc
     8041 gaggaagcgg aatatatcct gtatcacata ttctgctgac gcaccggtgc agcctttttt
     8101 ctcctgccac atgaagcact tcactgacac cctcatcagt gccaacatag taagccagta
     8161 tacactccgc tagcgctgat gtccggaatc gtcacacaaa aaaccaacac acagatgtaa
     8221 tgaaaataaa gatattttat tgcggccatc gtgatggcta gagtttagcc ctcccacaca
     8281 taaccagagg gcagcaattc acgaatccca actgccgtcg gctgtccatc actgtccttc
     8341 actatggctt tgatcccagg atgcagatcg agaagcacct gtcggcaccg tccgcagggg
     8401 ctcaagatgc ccctgttctc atttccgatc gcgacgatac aagtcaggtt gccagctgcc
     8461 gcagcagcag cagtgcccag caccacgagt tctgcacaag gtcccccagt aaaatgatat
     8521 acattgacac cagtgaagat gcggccgtcg ctagagagag ctgcgctggc gacgctgtag
     8581 tcttcagaga tggggatgct gttgattgta gccgttgctc tttcaatgag ggtggattct
     8641 tcttgagaca aaggcttggc catggtggcg ctagtgtcag aagaatcgag ctttttgcaa
     8701 aagcctaggc ctccaaaaaa gcctcctcac tacttctgga atagctcaga ggccgaggcg
     8761 gcctcggcct ctgcataaat aaaaaaaatt agtcagccat ggggcggaga atgggcggaa
     8821 ctgggcggag ttaggggcgg gatgggcgga gttaggggcg ggactatggt tgctgactaa
     8881 ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt tccacacctg
     8941 gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga gcctggggac
     9001 tttccacacc ctaactgaca cacattccac agctggttct ttccgcctca gaaggtacct
     9061 aaccaagttc ctctttcaga ggttatttca ggccatggtg ctgcgcaaga cgcgttaatt
     9121 aagacgt
//
