LOCUS       sequence-477408-        7305 bp    DNA     circular SYN 30-JUN-2025
DEFINITION  Express FBXO42-HaloTag(R) 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 7305)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 7305)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 30, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: FBXO42-HaloTag Vector
FEATURES             Location/Qualifiers
     source          1..7305
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(747..1541)
                     /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(858..877)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1468..1487
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(1573..1620)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     protein_bind    1630..1663
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    1778..1899
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     1869..1888
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(1986..2870)
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="EIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYV
                     WRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHD
                     WGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQ
                     NVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEE
                     YMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIG
                     SEIARWLSTLEISG"
     CDS             complement(2886..2906)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     primer_bind     complement(5091..5110)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5092..5110)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5152..5171)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     intron          5154..5286
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        complement(5422..5625)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(5451..5471)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        5626..5929
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    6162..6243
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        6322..6679
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6322..6513
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6345..6365)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6530..6665
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      6704..6713
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6710..7108
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    7132..7180
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 ttgaaatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccggttg
      721 ggcgtcgctt ggtcggtcat ttcgaatcag aagaactcgt caagaaggcg atagaaggcg
      781 atgcgctgcg aatcgggagc ggcgataccg taaagcacga ggaagcggtc agcccattcg
      841 ccgccaagct cttcagcaat atcacgggta gccaacgcta tgtcctgata gcggtccgcc
      901 acacccagcc ggccacagtc gatgaatcca gaaaagcggc cattttccac catgatattc
      961 ggcaagcagg catcgccatg ggtcacgacg agatcctcgc cgtcgggcat gcgcgccttg
     1021 agcctggcga acagttcggc tggcgcgagc ccctgatgct cttcgtccag atcatcctga
     1081 tcgacaagac cggcttccat ccgagtacgt gctcgctcga tgcgatgttt cgcttggtgg
     1141 tcgaatgggc aggtagccgg atcaagcgta tgcagccgcc gcattgcatc agccatgatg
     1201 gatactttct cggcaggagc aaggtgagat gacaggagat cctgccccgg cacttcgccc
     1261 aatagcagcc agtcccttcc cgcttcagtg acaacgtcga gcacagctgc gcaaggaacg
     1321 cccgtcgtgg ccagccacga tagccgcgct gcctcgtcct gcagttcatt cagggcaccg
     1381 gacaggtcgg tcttgacaaa aagaaccggg cgcccctgcg ctgacagccg gaacacggcg
     1441 gcatcagagc agccgattgt ctgttgtgcc cagtcatagc cgaatagcct ctccacccaa
     1501 gcggccggag aacctgcgtg caatccatct tgttcaatca tgatctgcct cctgggttta
     1561 gttcctcacc ttgtcgtatt atactatgcc gatatactat gccgatgatt aattgtcaac
     1621 agtggatcca taacttcgta taatgtatgc tatacgaagt tatattagaa ttcttaatta
     1681 agattttacc acatttgtag aggttttact tgcttaaaaa aacctcccac atctccccct
     1741 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa
     1801 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     1861 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgct cgaagcggcc
     1921 ttgctcagcg gtggcagcag ccaaatcagc ttgcatgcct gcaggtcgac tctagaattc
     1981 tattaaccgg aaatctccag agtagacagc cagcgcgcga tctcgctgcc gatcaggtcc
     2041 gggttgtctt cttgcagcag attcagaccc gggccgatgt ccacagcctt gcagttaggc
     2101 aggcttttgg ccaggcgagc ggcttcggcc ggtgggatca gaacgcctgg ggtgccccag
     2161 aacagcagct tcgggacagg ggactggtgc agccagtcca tgtattcttc gaccagcgcg
     2221 acgatgttcg ctggctcacc ggcgattggc agctcgtttg ggaagcgcca cagtggctcg
     2281 cggtcaacag gattcaggaa cggctcgcgg taatggtcca tctcgacttc agtcagcggg
     2341 cggacgacac ccatcggcag cgtaccctcg ataaaaacgt tctgatcgat gatcagcttg
     2401 cggccgacgt cggtggtgcg gaaggcctgg aaggtctcgc gggcaaattc tggccattcg
     2461 tcccaggtcg ggatagggcg gatgaactcc ataaatgcaa tacctttgac gcgctctgga
     2521 ttgcgcttgg cccagtggaa acccagagcg gagccccagt cgtgaatgac caggacgacc
     2581 tcttccagac ccagggcttc gatgaaggca tccatgaagc ggacgtggtc gtcgaagaaa
     2641 taacccaggt ctggtttgtc ggatttgccc ataccgatca ggtctggagc aatgcagcga
     2701 tgggtcggtg caacatgcgg gatgatgttg cgccacacgt aggaggaggt cgggttaccg
     2761 tgcaggaaca gcacaggggt gccatcgcgc ggaccaacat cgacgtagtg catgcgctcg
     2821 cccaggactt ccacataatg ggggtcgaat ggaaagccag taccgatttc ggatccatcg
     2881 ttatcgctct gaaagtacag atcctcagtg gttggctcga gagaaactct ctttgctcgt
     2941 acaaagtaca aggcgtttgt ttttggatag tacttcacat tctgtttctt gtccatgagt
     3001 cctccaaata tgatgagttc acccctgcct tgtaccacgg tatgcaggct ggtttcagga
     3061 ggtccaacca cagaactgct attaaatact ttccatttga cccgcccctt ctccttggtg
     3121 tctttaatgt ccagcacgta catctgcatg ggcttgcagt tcatactctg gtataggggt
     3181 ttgccaacat ttagggactg tggagggtgg tggcccaggc ggcgagcaat gggaggtaag
     3241 gaatgtccat ctccttgggc aggccctggg cgagggatgg gcactgtttc tccactgctc
     3301 aaactctggc tcccaggaga gcctggagaa gacccaagag gaggacttag tgctgcagag
     3361 gccgaggggc ctttggagga catcgctttg atggcttcca gactccgacg cagggcacct
     3421 ggggagacgg cccctgcaag ggcactggcc acgtgaggtg gggtatgcac accatttgtc
     3481 tgttcaggag ggtgtctcat acttccccca actgtcctat tgtccatgcc atccatggga
     3541 ttactactgg aagcgggttt cagatcccaa ttcagatcta tggatcctaa tctcagatct
     3601 ttctgatctg gtagtgatcc tcgtcggggg gccaaagaaa gtcctatttt caggtcgtat
     3661 ccttcaggag cagatggagt acttggagat atggcctgta caggactgtc caaagaagag
     3721 ccacccacag ctgccgttcc tggagacaaa ctcccaccat tgaggatagg agagccgtct
     3781 cctctggctg gggaaaggct cccttcccgg gaacctgaag gagtctgcct ttgagccctg
     3841 ggtctcagtg ttccccagcg gccgttaaca caaggagctt catccatgct tcttactgga
     3901 gactgagagc ggtactctcg ggtttcagga acgagagctg gaggagtggc actgataggt
     3961 gatgggcgag agttcaaact ggggctgagt ggggctctcc cactaggagc ctggctgaag
     4021 accaccacac actgtcccac ccggcaagct ggatggcacc acagttctgg ggccccatgc
     4081 tcttcatttt ctaccttgag tggctgccag gcccaaggac cagaatgcat gtgcaacaac
     4141 caagcatcct tgaatagagc attgggaccg ccacaccctc cgaggattaa gatagttgca
     4201 tcatctatga caatctgaga ttggccacct cgaggatgag gactggggcc agagatgttc
     4261 ggcttggacc acgcccactg ctcaaggtca aggacccaga catcattgct catttgccgg
     4321 gatcctaaag agccaccaaa gacaatcatt ttatcatcta tcacacagga ggagtggcca
     4381 gccatgggag gtggcccatg ggttgtcaca atgcagttcc accaattttt agagggtgag
     4441 taagtgtgta tttcatcaaa gaatctctct ggctggtgta ggggataagg gcttggccgc
     4501 gtccagccac caaacagcac tagcaagtcc ttgtacacga ccagagttgc tccagctttg
     4561 ggggaaggat aggaccctga agccaaaggt cggatccact ctttgctgtt taggtcaagt
     4621 ctccagaggt cattgaaagc agcattgcag ctgctctggg tacagcctcc aaacacatac
     4681 atagactgat tagcatcata atagcatgca ctgtgcgaga agcgctgagt gattggggtt
     4741 ccaggataag gataggtacg gctctcccac tgaatgtttc cttcctggac agccttcatg
     4801 aaaccatgat aacactgatg ggctacacct ttgataagtc gataccactg tttgcagaca
     4861 agggccgcag ttttgtgttc ctgatacggt gagagaaagg acaggatata ctccaaaacc
     4921 tcttctggca gctccgacat ggacctatta tgtctagtct cctcagcctc caatactggg
     4981 tggggctcct catcttgatc cattgtccct tccagcacag tttcttcctg gtccacagcc
     5041 atgaaactgt catcttcact gtccgaggag ctggccatgg cgatcgctag ccctatagtg
     5101 agtcgtatta atactctagc cttaagagct gtaattgaac tgggagtgga cacctgtgga
     5161 gagaaaggca aagtggatgt cagtaagacc aataggtgcc tatcagaaac gcaagagtct
     5221 tctctgtctc gacaagccca gtttctattg gtctccttaa acctgtcttg taaccttgat
     5281 acttacctgc ccagtgcctc acgaccaact tctgcagctt aagttcgaga ctgttgtgtc
     5341 agaagcactg actgcgttag caatttaact gtgataaact accgcaataa agcttctagt
     5401 gatctgacgg ttcactaaac cagctctgct tatatagacc tcccaccgta cacgcctacc
     5461 gcccatttgc gtcaacgggg cggggttatt acgacatttt ggaaagtccc gttgattttg
     5521 gtgccaaaac aaactcccat tgacgtcaat ggggtggaga cttggaaatc cccgtgagtc
     5581 aaaccgctat ccacgcccat tggtgtactg ccaaaaccgc atcaccatgg taatagcgat
     5641 gactaatacg tagatgtact gccaagtagg aaagtcccgt aaggtcatgt actgggcata
     5701 atgccaggcg ggccatttac cgtcattgac gtcaataggg ggcggacttg gcatatgata
     5761 cacttgatgt actgccaagt gggcagttta ccgtaaatac tccacccatt gacgtcaatg
     5821 gaaagtccct attggcgtta ctatgggaac atacgtcatt attgacgtca atgggcgggg
     5881 gtcgttgggc ggtcagccag gcgggccatt taccgtaagt tatgtaacgc ggaactccat
     5941 atatgggcta tgaactaatg accccgtaat tgattactat taataactag tggcgcgcca
     6001 taatcctagg ctctcgtacg aagtcagctg ataacgtcga cgtattgcgg ccgcggatca
     6061 taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc
     6121 ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt
     6181 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     6241 tgccgacgtc ttaattaacg cgtcttgcgc agcaccatgg cctgaaataa cctctgaaag
     6301 aggaacttgg ttaggtacct tctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt
     6361 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     6421 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     6481 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     6541 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     6601 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     6661 aggcctaggc ttttgcaaaa agctcgattc ttctgacact agcgccacca tggccaagcc
     6721 tttgtctcaa gaagaatcca ccctcattga aagagcaacg gctacaatca acagcatccc
     6781 catctctgaa gactacagcg tcgccagcgc agctctctct agcgacggcc gcatcttcac
     6841 tggtgtcaat gtatatcatt ttactggggg accttgtgca gaactcgtgg tgctgggcac
     6901 tgctgctgct gcggcagctg gcaacctgac ttgtatcgtc gcgatcggaa atgagaacag
     6961 gggcatcttg agcccctgcg gacggtgccg acaggtgctt ctcgatctgc atcctgggat
     7021 caaagccata gtgaaggaca gtgatggaca gccgacggca gttgggattc gtgaattgct
     7081 gccctctggt tatgtgtggg agggctaaac tctagccatc acgatggccg caataaaata
     7141 tctttatttt cattacatct gtgtgttggt tttttgtgtg acgattccgg actataaaag
     7201 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     7261 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttc
//
