LOCUS       sequence-474689-        7708 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express HaloTag(R)-TTC7B 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 7708)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7708)
  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            "Unpublished"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: HaloTag-TTC7B Fusion Vector
FEATURES             Location/Qualifiers
     source          1..7708
                     /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    4750..4871
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4761..4780)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(4986..5019)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        5029..5076
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5108..5902
                     /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(5162..5181)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5772..5791
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(6060..6648)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6140..6159)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    6774..6822
                     /note="synthetic polyadenylation signal"
     CDS             complement(6846..7244)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     promoter        complement(7275..7632)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7289..7424
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        7441..7632
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     7589..7609
                     /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 ggcgaccaag aaggcaggct cgcggctgga gacggagatc gagcgctgcc
     2161 gctccgagtg ccagtgggag cggatccctg agctcgtcaa gcagctgtcg gccaagctca
     2221 tcgccaacga tgacatggca gagcttctcc tcggggagtc gaagctggag cagtacctga
     2281 aggaacaccc cctgaggcag ggggccagtc cccgaggccc caagccccag ctgactgagg
     2341 tccgcaagca tctgaccgcc gccctggacc gagggaacct taagtcagaa ttcctacaag
     2401 aatccaatct gatcatggcc aagttgaatt atgtggaagg tgattataaa gaagctctga
     2461 acatttacgc ccgggtgggc ctggacgatc tgccactgac agctgtcccg ccctacaggc
     2521 tgcgggtgat cgcagaagcc tacgctacca aaggactttg tttggagaag ctgcctattt
     2581 cttcttctac cagtaatctc catgtggacc gggaacagga tgtcatcacc tgttatgaga
     2641 aagcagggga catcgcactc ctgtatctcc aagagataga aagggtaata ctttctaata
     2701 ttcaaaacag aagccctaag cctggccctg ctccccacga tcaagaacta ggttttttcc
     2761 tagaaacagg acttcagaga gcccatgtcc tctatttcaa aaatgggaac ttgacaagag
     2821 gagtcggaag atttagagag cttctcagag cagttgaaac aagaacgact caaaacctgc
     2881 gaatgacaat agccaggcag ctggcagaga tcttgttgcg gggtatgtgt gagcagagct
     2941 actggaaccc tctggaggat ccaccgtgcc agtcacctct ggacgatcct ctccgcaaag
     3001 gagcaaacac aaaaacctac actctcactc ggagagcccg tgtctactca ggagagaaca
     3061 ttttttgtcc tcaagaaaat acggaagaag ccctgttgtt attgctgatt agtgaatcaa
     3121 tggccaaccg ggacgctgtg ctgagcagga tacctgaaca caagagtgac cgcctcatca
     3181 gtctgcagag tgcatctgtg gtctatgact tactcaccat tgctcttgga agaagaggcc
     3241 agtatgagat gctgtcagag tgcctagaaa gagccatgaa gtttgccttt gaggaattcc
     3301 acctgtggta ccagtttgct ctgtccctga tggctgctgg aaaatctgcc cgtgccgtga
     3361 aggtgctgaa agagtgtatc cgcctgaagc cagacgatgc caccatccct ctcctcgctg
     3421 ccaagctctg catgggctcc ctgcactggt tggaagaggc tgaaaagttt gccaaaactg
     3481 tcgttgatgt gggagagaaa acgtcagagt tcaaggccaa aggctactta gctctggggc
     3541 tcacgtacag tctgcaggcc actgacgctt ctttgcgagg gatgcaggag gtcctacaga
     3601 gaaaggcgct tcttgcattt cagagggccc acagcctgtc acccacagat caccaagcag
     3661 ctttctacct ggctctgcag cttgccatct ccagacagat cccagaggct ctggggtatg
     3721 tccgccaagc tcttcagctt caaggtgacg atgccaactc cctgcacctc cttgccctcc
     3781 tgctgtcagc acagaagcat taccatgacg ctctgaacat catcgacatg gccctgagtg
     3841 aatacccaga aaatttcata ctactgtttt ccaaagtgaa gttgcagtca ctctgccgag
     3901 gcccggacga ggcactgctg acttgtaagc acatgctgca gatatggaaa tcctgctaca
     3961 acctcaccaa ccccagtgat tctggacgtg ggagcagcct cttagataga accattgctg
     4021 acagacgaca gcttaataca attactttgc cagacttcag cgatcccgag acaggctccg
     4081 tccatgccac atcggtagca gcctcaagag tggagcaggc actgtcggaa gtggcttcgt
     4141 ctctgcagag cagtgcccct aagcagggcc cgctgcaccc ctggatgacg ctggcacaga
     4201 tctggctcca tgcagctgaa gtctatatcg gcatcgggaa gcctgcagaa gccacagcct
     4261 gtacccaaga agctgccaac ctcttcccaa tgtcccacaa tgtcctctac atgcgcggcc
     4321 agattgctga gctccgggga agcatggacg aggcgcggcg gtggtatgaa gaggccttag
     4381 ccatcagccc cacccacgtg aagagcatgc agcgactggc cctgatcctt caccagctag
     4441 gccgctacag tctggcggag aagatcctcc gggacgcggt gcaggtgaac tcgacagccc
     4501 acgaggtctg gaacgggctg ggcgaggtcc tccaagctca gggcaacgat gcggcggcta
     4561 cggagtgctt cctgacagcc ttggagctgg aggccagcag ccccgccgtg cccttcacca
     4621 tcatcccccg cgtgctcgtt taaacgaatt cgggctcggt acccggggat cctctagagt
     4681 cgacctgcag gcatgcaagc tgatttggct gctgccaccg ctgagcaagg ccgcttcgag
     4741 cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa
     4801 aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca
     4861 ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag ggggagatgt
     4921 gggaggtttt tttaagcaag taaaacctct acaaatgtgg taaaatctta attaagaatt
     4981 ctaatataac ttcgtatagc atacattata cgaagttatg gatccactgt tgacaattaa
     5041 tcatcggcat agtatatcgg catagtataa tacgacaagg tgaggaacta aacccaggag
     5101 gcagatcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     5161 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     5221 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     5281 tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     5341 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     5401 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     5461 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     5521 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     5581 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat
     5641 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     5701 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     5761 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     5821 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     5881 ccttcttgac gagttcttct gattcgaaat gaccgaccaa gcgacgccca accggtatca
     5941 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     6001 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     6061 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     6121 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     6181 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     6241 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     6301 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     6361 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     6421 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     6481 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc
     6541 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     6601 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatttcaaga agatcctttg
     6661 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     6721 atgagattat caaaaaggat cttcacctag atccttttat agtccggaat cgtcacacaa
     6781 aaaaccaaca cacagatgta atgaaaataa agatatttta ttgcggccat cgtgatggct
     6841 agagtttagc cctcccacac ataaccagag ggcagcaatt cacgaatccc aactgccgtc
     6901 ggctgtccat cactgtcctt cactatggct ttgatcccag gatgcagatc gagaagcacc
     6961 tgtcggcacc gtccgcaggg gctcaagatg cccctgttct catttccgat cgcgacgata
     7021 caagtcaggt tgccagctgc cgcagcagca gcagtgccca gcaccacgag ttctgcacaa
     7081 ggtcccccag taaaatgata tacattgaca ccagtgaaga tgcggccgtc gctagagaga
     7141 gctgcgctgg cgacgctgta gtcttcagag atggggatgc tgttgattgt agccgttgct
     7201 ctttcaatga gggtggattc ttcttgagac aaaggcttgg ccatggtagc gctagtgtca
     7261 gaagaatcga gctttttgca aaagcctagg cctccaaaaa agcctcctca ctacttctgg
     7321 aatagctcag aggccgaggc ggcctcggcc tctgcataaa taaaaaaaat tagtcagcca
     7381 tggggcggag aatgggcgga actgggcgga gttaggggcg ggatgggcgg agttaggggc
     7441 gggactatgg ttgctgacta attgagatgc atgctttgca tacttctgcc tgctggggag
     7501 cctggggact ttccacacct ggttgctgac taattgagat gcatgctttg catacttctg
     7561 cctgctgggg agcctgggga ctttccacac cctaactgac acacattcca cagctggttc
     7621 tttccgcctc agaaggtacc taaccaagtt cctctttcag aggttatttc aggccatggt
     7681 gctgcgcaag acgcgttaat taagacgt
//
