LOCUS       sequence-476798-        7330 bp    DNA     circular SYN 20-JUN-2025
DEFINITION  Express HaloTag(R)-FBXO42 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 7330)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 7330)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 20, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: HaloTag-FBXO42 Vector
FEATURES             Location/Qualifiers
     source          1..7330
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    145..226
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        305..662
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        305..496
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(328..348)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      513..648
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      687..696
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             693..1091
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    1115..1163
                     /note="synthetic polyadenylation signal"
     rep_origin      1289..1877
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1778..1797
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2035..2829)
                     /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(2146..2165)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2756..2775
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(2861..2908)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     protein_bind    2918..2951
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    3066..3187
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     3157..3176
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(5466..5486)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             complement(5499..6389)
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
     regulatory      6386..6395
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(6404..6423)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6405..6423)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6465..6484)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     intron          6467..6599
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        complement(6735..6938)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(6764..6784)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        6939..7242
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 acgaagtcag ctgataacgt cgacgtattg cggccgcgga tcataatcag ccataccaca
       61 tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat
      121 aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa
      181 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgccgac gtcttaatta
      241 acgcgtcttg cgcagcacca tggcctgaaa taacctctga aagaggaact tggttaggta
      301 ccttctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc
      361 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
      421 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
      481 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
      541 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
      601 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
      661 aaaagctcga ttcttctgac actagcgcca ccatggccaa gcctttgtct caagaagaat
      721 ccaccctcat tgaaagagca acggctacaa tcaacagcat ccccatctct gaagactaca
      781 gcgtcgccag cgcagctctc tctagcgacg gccgcatctt cactggtgtc aatgtatatc
      841 attttactgg gggaccttgt gcagaactcg tggtgctggg cactgctgct gctgcggcag
      901 ctggcaacct gacttgtatc gtcgcgatcg gaaatgagaa caggggcatc ttgagcccct
      961 gcggacggtg ccgacaggtg cttctcgatc tgcatcctgg gatcaaagcc atagtgaagg
     1021 acagtgatgg acagccgacg gcagttggga ttcgtgaatt gctgccctct ggttatgtgt
     1081 gggagggcta aactctagcc atcacgatgg ccgcaataaa atatctttat tttcattaca
     1141 tctgtgtgtt ggttttttgt gtgacgattc cggactataa aaggatctag gtgaagatcc
     1201 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     1261 accccgtaga aaagatcaaa ggatcttctt gaaatccttt ttttctgcgc gtaatctgct
     1321 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     1381 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     1441 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     1501 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     1561 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     1621 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     1681 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     1741 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     1801 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     1861 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     1921 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     1981 ccgcctttga gtgagctgat accggttggg cgtcgcttgg tcggtcattt cgaatcagaa
     2041 gaactcgtca agaaggcgat agaaggcgat gcgctgcgaa tcgggagcgg cgataccgta
     2101 aagcacgagg aagcggtcag cccattcgcc gccaagctct tcagcaatat cacgggtagc
     2161 caacgctatg tcctgatagc ggtccgccac acccagccgg ccacagtcga tgaatccaga
     2221 aaagcggcca ttttccacca tgatattcgg caagcaggca tcgccatggg tcacgacgag
     2281 atcctcgccg tcgggcatgc gcgccttgag cctggcgaac agttcggctg gcgcgagccc
     2341 ctgatgctct tcgtccagat catcctgatc gacaagaccg gcttccatcc gagtacgtgc
     2401 tcgctcgatg cgatgtttcg cttggtggtc gaatgggcag gtagccggat caagcgtatg
     2461 cagccgccgc attgcatcag ccatgatgga tactttctcg gcaggagcaa ggtgagatga
     2521 caggagatcc tgccccggca cttcgcccaa tagcagccag tcccttcccg cttcagtgac
     2581 aacgtcgagc acagctgcgc aaggaacgcc cgtcgtggcc agccacgata gccgcgctgc
     2641 ctcgtcctgc agttcattca gggcaccgga caggtcggtc ttgacaaaaa gaaccgggcg
     2701 cccctgcgct gacagccgga acacggcggc atcagagcag ccgattgtct gttgtgccca
     2761 gtcatagccg aatagcctct ccacccaagc ggccggagaa cctgcgtgca atccatcttg
     2821 ttcaatcatg atctgcctcc tgggtttagt tcctcacctt gtcgtattat actatgccga
     2881 tatactatgc cgatgattaa ttgtcaacag tggatccata acttcgtata atgtatgcta
     2941 tacgaagtta tattagaatt cttaattaag attttaccac atttgtagag gttttacttg
     3001 cttaaaaaaa cctcccacat ctccccctga acctgaaaca taaaatgaat gcaattgttg
     3061 ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     3121 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     3181 tatcttatca tgtctgctcg aagcggcctt gctcagcggt ggcagcagcc aaatcagctt
     3241 gcatgcctgc aggtcgactc tagaggatcc ccgggtaccg agcccgaatt cgtttaaact
     3301 ctctttgctc gtacaaagta caaggcgttt gtttttggat agtacttcac attctgtttc
     3361 ttgtccatga gtcctccaaa tatgatgagt tcacccctgc cttgtaccac ggtatgcagg
     3421 ctggtttcag gaggtccaac cacagaactg ctattaaata ctttccattt gacccgcccc
     3481 ttctccttgg tgtctttaat gtccagcacg tacatctgca tgggcttgca gttcatactc
     3541 tggtataggg gtttgccaac atttagggac tgtggagggt ggtggcccag gcggcgagca
     3601 atgggaggta aggaatgtcc atctccttgg gcaggccctg ggcgagggat gggcactgtt
     3661 tctccactgc tcaaactctg gctcccagga gagcctggag aagacccaag aggaggactt
     3721 agtgctgcag aggccgaggg gcctttggag gacatcgctt tgatggcttc cagactccga
     3781 cgcagggcac ctggggagac ggcccctgca agggcactgg ccacgtgagg tggggtatgc
     3841 acaccatttg tctgttcagg agggtgtctc atacttcccc caactgtcct attgtccatg
     3901 ccatccatgg gattactact ggaagcgggt ttcagatccc aattcagatc tatggatcct
     3961 aatctcagat ctttctgatc tggtagtgat cctcgtcggg gggccaaaga aagtcctatt
     4021 ttcaggtcgt atccttcagg agcagatgga gtacttggag atatggcctg tacaggactg
     4081 tccaaagaag agccacccac agctgccgtt cctggagaca aactcccacc attgaggata
     4141 ggagagccgt ctcctctggc tggggaaagg ctcccttccc gggaacctga aggagtctgc
     4201 ctttgagccc tgggtctcag tgttccccag cggccgttaa cacaaggagc ttcatccatg
     4261 cttcttactg gagactgaga gcggtactct cgggtttcag gaacgagagc tggaggagtg
     4321 gcactgatag gtgatgggcg agagttcaaa ctggggctga gtggggctct cccactagga
     4381 gcctggctga agaccaccac acactgtccc acccggcaag ctggatggca ccacagttct
     4441 ggggccccat gctcttcatt ttctaccttg agtggctgcc aggcccaagg accagaatgc
     4501 atgtgcaaca accaagcatc cttgaataga gcattgggac cgccacaccc tccgaggatt
     4561 aagatagttg catcatctat gacaatctga gattggccac ctcgaggatg aggactgggg
     4621 ccagagatgt tcggcttgga ccacgcccac tgctcaaggt caaggaccca gacatcattg
     4681 ctcatttgcc gggatcctaa agagccacca aagacaatca ttttatcatc tatcacacag
     4741 gaggagtggc cagccatggg aggtggccca tgggttgtca caatgcagtt ccaccaattt
     4801 ttagagggtg agtaagtgtg tatttcatca aagaatctct ctggctggtg taggggataa
     4861 gggcttggcc gcgtccagcc accaaacagc actagcaagt ccttgtacac gaccagagtt
     4921 gctccagctt tgggggaagg ataggaccct gaagccaaag gtcggatcca ctctttgctg
     4981 tttaggtcaa gtctccagag gtcattgaaa gcagcattgc agctgctctg ggtacagcct
     5041 ccaaacacat acatagactg attagcatca taatagcatg cactgtgcga gaagcgctga
     5101 gtgattgggg ttccaggata aggataggta cggctctccc actgaatgtt tccttcctgg
     5161 acagccttca tgaaaccatg ataacactga tgggctacac ctttgataag tcgataccac
     5221 tgtttgcaga caagggccgc agttttgtgt tcctgatacg gtgagagaaa ggacaggata
     5281 tactccaaaa cctcttctgg cagctccgac atggacctat tatgtctagt ctcctcagcc
     5341 tccaatactg ggtggggctc ctcatcttga tccattgtcc cttccagcac agtttcttcc
     5401 tggtccacag ccatgaaact gtcatcttca ctgtccgagg agctggccat ggcgatcgcg
     5461 ttatcgctct gaaagtacag atcctcagtg gttggctcgc cggaaatctc gagcgtcgac
     5521 agccagcgcg cgatctcgct gccgatcagg tccgggttgt cttcttgcag cagattcaga
     5581 cccgggccga tgtccacagc cttgcagtta ggcaggcttt tggccaggcg agcggcttcg
     5641 gccggtggga tcagaacgcc tggggtgccc cagaacagca gcttcgggac aggggactgg
     5701 tgcagccagt ccatgtattc ttcgaccagc gcgacgatgt tcgctggctc accggcgatt
     5761 ggcagctcgt ttgggaagcg ccacagtggc tcgcggtcaa caggattcag gaacggctcg
     5821 cggtaatggt ccatctcgac ttcagtcagc gggcggacga cacccatcgg cagcgtaccc
     5881 tcgataaaaa cgttctgatc gatgatcagc ttgcggccga cgtcggtggt gcggaaggcc
     5941 tggaaggtct cgcgggcaaa ttctggccat tcgtcccagg tcgggatagg gcggatgaac
     6001 tccataaatg caataccttt gacgcgctct ggattgcgct tggcccagtg gaaacccaga
     6061 gcggagcccc agtcgtgaat gaccaggacg acctcttcca gacccagggc ttcgatgaag
     6121 gcatccatga agcggacgtg gtcgtcgaag aaataaccca ggtctggttt gtcggatttg
     6181 cccataccga tcaggtctgg agcaatgcag cgatgggtcg gtgcaacatg cgggatgatg
     6241 ttgcgccaca cgtaggagga ggtcgggtta ccgtgcagga acagcacagg ggtgccatcg
     6301 cgcggaccaa catcgacgta gtgcatgcgc tcgcccagga cttccacata atgggggtcg
     6361 aatggaaagc cagtaccgat ttctgccatg gtggctttgc tagccctata gtgagtcgta
     6421 ttaatactct agccttaaga gctgtaattg aactgggagt ggacacctgt ggagagaaag
     6481 gcaaagtgga tgtcagtaag accaataggt gcctatcaga aacgcaagag tcttctctgt
     6541 ctcgacaagc ccagtttcta ttggtctcct taaacctgtc ttgtaacctt gatacttacc
     6601 tgcccagtgc ctcacgacca acttctgcag cttaagttcg agactgttgt gtcagaagca
     6661 ctgactgcgt tagcaattta actgtgataa actaccgcaa taaagcttct agtgatctga
     6721 cggttcacta aaccagctct gcttatatag acctcccacc gtacacgcct accgcccatt
     6781 tgcgtcaacg gggcggggtt attacgacat tttggaaagt cccgttgatt ttggtgccaa
     6841 aacaaactcc cattgacgtc aatggggtgg agacttggaa atccccgtga gtcaaaccgc
     6901 tatccacgcc cattggtgta ctgccaaaac cgcatcacca tggtaatagc gatgactaat
     6961 acgtagatgt actgccaagt aggaaagtcc cgtaaggtca tgtactgggc ataatgccag
     7021 gcgggccatt taccgtcatt gacgtcaata gggggcggac ttggcatatg atacacttga
     7081 tgtactgcca agtgggcagt ttaccgtaaa tactccaccc attgacgtca atggaaagtc
     7141 cctattggcg ttactatggg aacatacgtc attattgacg tcaatgggcg ggggtcgttg
     7201 ggcggtcagc caggcgggcc atttaccgta agttatgtaa cgcggaactc catatatggg
     7261 ctatgaacta atgaccccgt aattgattac tattaataac tagtggcgcg ccataatcct
     7321 aggctctcgt
//
