LOCUS       sequence-479887-        7803 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Express IL4Ra-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 7803)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7803)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 7, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: IL4Ra-HaloTag BiBRET Vector
FEATURES             Location/Qualifiers
     source          1..7803
                     /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             3664..3684
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             3700..4584
                     /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    4671..4792
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4682..4701)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(4907..4940)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        4950..4997
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5029..5823
                     /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(5083..5102)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5693..5712
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      6040..6585
                     /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     6486..6505
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6702..6719
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    6869..6917
                     /note="synthetic polyadenylation signal"
     CDS             complement(6941..7339)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      7336..7345
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(7370..7727)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7384..7519
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        7536..7727
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     7684..7704
                     /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 gggtggcttt gctctgggct cctgttccct
     1201 gtgagctgcc tggtcctgct gcaggtggca agctctggga acatgaaggt cttgcaggag
     1261 cccacctgcg tctccgacta catgagcatc tctacttgcg agtggaagat gaatggtccc
     1321 accaattgca gcaccgagct ccgcctgttg taccagctgg tttttctgct ctccgaagcc
     1381 cacacgtgta tccctgagaa caacggaggc gcggggtgcg tgtgccacct gctcatggat
     1441 gacgtggtca gtgcggataa ctatacactg gacctgtggg ctgggcagca gctgctgtgg
     1501 aagggctcct tcaagcccag cgagcatgtg aaacccaggg ccccaggaaa cctgacagtt
     1561 cacaccaatg tctccgacac tctgctgctg acctggagca acccgtatcc ccctgacaat
     1621 tacctgtata atcatctcac ctatgcagtc aacatttgga gtgaaaacga cccggcagat
     1681 ttcagaatct ataacgtgac ctacctagaa ccctccctcc gcatcgcagc cagcaccctg
     1741 aagtctggga tttcctacag ggcacgggtg agggcctggg ctcagtgcta taacaccacc
     1801 tggagtgagt ggagccccag caccaagtgg cacaactcct acagggagcc cttcgagcag
     1861 cacctcctgc tgggcgtcag cgtttcctgc attgtcatcc tggccgtctg cctgttgtgc
     1921 tatgtcagca tcaccaagat taagaaagaa tggtgggatc agattcccaa cccagcccgc
     1981 agccgcctcg tggctataat aatccaggat gctcaggggt cacagtggga gaagcggtcc
     2041 cgaggccagg aaccagccaa gtgcccacac tggaagaatt gtcttaccaa gctcttgccc
     2101 tgttttctgg agcacaacat gaaaagggat gaagatcctc acaaggctgc caaagagatg
     2161 cctttccagg gctctggaaa atcagcatgg tgcccagtgg agatcagcaa gacagtcctc
     2221 tggccagaga gcatcagcgt ggtgcgatgt gtggagttgt ttgaggcccc ggtggagtgt
     2281 gaggaggagg aggaggtaga ggaagaaaaa gggagcttct gtgcatcgcc tgagagcagc
     2341 agggatgact tccaggaggg aagggagggc attgtggccc ggctaacaga gagcctgttc
     2401 ctggacctgc tcggagagga gaatgggggc ttttgccagc aggacatggg ggagtcatgc
     2461 cttcttccac cttcgggaag tacgagtgct cacatgccct gggatgagtt cccaagtgca
     2521 gggcccaagg aggcacctcc ctggggcaag gagcagcctc tccacctgga gccaagtcct
     2581 cctgccagcc cgacccagag tccagacaac ctgacttgca cagagacgcc cctcgtcatc
     2641 gcaggcaacc ctgcttaccg cagcttcagc aactccctga gccagtcacc gtgtcccaga
     2701 gagctgggtc cagacccact gctggccaga cacctggagg aagtagaacc cgagatgccc
     2761 tgtgtccccc agctctctga gccaaccact gtgccccaac ctgagccaga aacctgggag
     2821 cagatcctcc gccgaaatgt cctccagcat ggggcagctg cagcccccgt ctcggccccc
     2881 accagtggct atcaggagtt tgtacatgcg gtggagcagg gtggcaccca ggccagtgcg
     2941 gtggtgggct tgggtccccc aggagaggct ggttacaagg ccttctcaag cctgcttgcc
     3001 agcagtgctg tgtccccaga gaaatgtggg tttggggcta gcagtgggga agaggggtat
     3061 aagcctttcc aagacctcat tcctggctgc cctggggacc ctgccccagt ccctgtcccc
     3121 ttgttcacct ttggactgga cagggagcca cctcgcagtc cgcagagctc acatctccca
     3181 agcagctccc cagagcacct gggtctggag ccgggggaaa aggtagagga catgccaaag
     3241 cccccacttc cccaggagca ggccacagac ccccttgtgg acagcctggg cagtggcatt
     3301 gtctactcag cccttacctg ccacctgtgc ggccacctga aacagtgtca tggccaggag
     3361 gatggtggcc agacccctgt catggccagt ccttgctgtg gctgctgctg tggagacagg
     3421 tcctcgcccc ctacaacccc cctgagggcc ccagacccct ctccaggtgg ggttccactg
     3481 gaggccagtc tgtgtccggc ctccctggca ccctcgggca tctcagagaa gagtaaatcc
     3541 tcatcatcct tccatcctgc ccctggcaat gctcagagct caagccagac ccccaaaatc
     3601 gtgaactttg tctccgtggg acccacatac atgagggtct ctgtttctct cgagccaacc
     3661 actgaggatc tgtactttca gagcgataac gatggatccg aaatcggtac tggctttcca
     3721 ttcgaccccc attatgtgga agtcctgggc gagcgcatgc actacgtcga tgttggtccg
     3781 cgcgatggca cccctgtgct gttcctgcac ggtaacccga cctcctccta cgtgtggcgc
     3841 aacatcatcc cgcatgttgc accgacccat cgctgcattg ctccagacct gatcggtatg
     3901 ggcaaatccg acaaaccaga cctgggttat ttcttcgacg accacgtccg cttcatggat
     3961 gccttcatcg aagccctggg tctggaagag gtcgtcctgg tcattcacga ctggggctcc
     4021 gctctgggtt tccactgggc caagcgcaat ccagagcgcg tcaaaggtat tgcatttatg
     4081 gagttcatcc gccctatccc gacctgggac gaatggccag aatttgcccg cgagaccttc
     4141 caggccttcc gcaccaccga cgtcggccgc aagctgatca tcgatcagaa cgtttttatc
     4201 gagggtacgc tgccgatggg tgtcgtccgc ccgctgactg aagtcgagat ggaccattac
     4261 cgcgagccgt tcctgaatcc tgttgaccgc gagccactgt ggcgcttccc aaacgagctg
     4321 ccaatcgccg gtgagccagc gaacatcgtc gcgctggtcg aagaatacat ggactggctg
     4381 caccagtccc ctgtcccgaa gctgctgttc tggggcaccc caggcgttct gatcccaccg
     4441 gccgaagccg ctcgcctggc caaaagcctg cctaactgca aggctgtgga catcggcccg
     4501 ggtctgaatc tgctgcaaga agacaacccg gacctgatcg gcagcgagat cgcgcgctgg
     4561 ctgtctactc tggagatttc cggttaatag aattctagag tcgacctgca ggcatgcaag
     4621 ctgatttggc tgctgccacc gctgagcaag gccgcttcga gcagacatga taagatacat
     4681 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     4741 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
     4801 caattgcatt cattttatgt ttcaggttca gggggagatg tgggaggttt ttttaagcaa
     4861 gtaaaacctc tacaaatgtg gtaaaatctt aattaagaat tctaatataa cttcgtatag
     4921 catacattat acgaagttat ggatccactg ttgacaatta atcatcggca tagtatatcg
     4981 gcatagtata atacgacaag gtgaggaact aaacccagga ggcagatcat gattgaacaa
     5041 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     5101 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     5161 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     5221 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     5281 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     5341 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     5401 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     5461 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     5521 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     5581 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     5641 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     5701 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     5761 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     5821 tgattcgaaa tgaccgacca agcgacgccc aaccggtatc agctcactca aaggcggtaa
     5881 tacggttatc cacagaatca ggggataacg caggaaagaa catgtcaaac atgagaatta
     5941 caacttatat cgtatggggc tgacttcagg tgctacattt gaagagataa attgcactga
     6001 aatctagaaa tattttatct gattaataag atgatcttct tgagatcgtt ttggtctgcg
     6061 cgtaatctct tgctctgaaa acgaaaaaac cgccttgcag ggcggttttt cgaaggttct
     6121 ctgagctacc aactctttga accgaggtaa ctggcttgga ggagcgcagt caccaaaact
     6181 tgtcctttca gtttagcctt aaccggcgca tgacttcaag actaactcct ctaaatcaat
     6241 taccagtggc tgctgccagt ggtgcttttg catgtctttc cgggttggac tcaagacgat
     6301 agttaccgga taaggcgcag cggtcggact gaacgggggg ttcgtgcata cagtccagct
     6361 tggagcgaac tgcctacccg gaactgagtg tcaggcgtgg aatgagacaa acgcggccat
     6421 aacagcggaa tgacaccggt aaaccgaaag gcaggaacag gagagcgcac gagggagccg
     6481 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacca ctgatttgag
     6541 cgtcagattt cgtgatgctt gtcagggggg cggagcctat ggaaaaacgg ctttgccgcg
     6601 gccctctcac ttccctgtta agtatcttcc tggcatcttc caggaaatct ccgccccgtt
     6661 cgtaagccat ttccgctcgc cgcagtcgaa cgaccgagcg tagcgagtca gtgagcgagg
     6721 aagcggaata tatcctgtat cacatattct gctgacgcac cggtgcagcc ttttttctcc
     6781 tgccacatga agcacttcac tgacaccctc atcagtgcca acatagtaag ccagtataca
     6841 ctccgctagc gctgatgtcc ggaatcgtca cacaaaaaac caacacacag atgtaatgaa
     6901 aataaagata ttttattgcg gccatcgtga tggctagagt ttagccctcc cacacataac
     6961 cagagggcag caattcacga atcccaactg ccgtcggctg tccatcactg tccttcacta
     7021 tggctttgat cccaggatgc agatcgagaa gcacctgtcg gcaccgtccg caggggctca
     7081 agatgcccct gttctcattt ccgatcgcga cgatacaagt caggttgcca gctgccgcag
     7141 cagcagcagt gcccagcacc acgagttctg cacaaggtcc cccagtaaaa tgatatacat
     7201 tgacaccagt gaagatgcgg ccgtcgctag agagagctgc gctggcgacg ctgtagtctt
     7261 cagagatggg gatgctgttg attgtagccg ttgctctttc aatgagggtg gattcttctt
     7321 gagacaaagg cttggccatg gtggcgctag tgtcagaaga atcgagcttt ttgcaaaagc
     7381 ctaggcctcc aaaaaagcct cctcactact tctggaatag ctcagaggcc gaggcggcct
     7441 cggcctctgc ataaataaaa aaaattagtc agccatgggg cggagaatgg gcggaactgg
     7501 gcggagttag gggcgggatg ggcggagtta ggggcgggac tatggttgct gactaattga
     7561 gatgcatgct ttgcatactt ctgcctgctg gggagcctgg ggactttcca cacctggttg
     7621 ctgactaatt gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc
     7681 cacaccctaa ctgacacaca ttccacagct ggttctttcc gcctcagaag gtacctaacc
     7741 aagttcctct ttcagaggtt atttcaggcc atggtgctgc gcaagacgcg ttaattaaga
     7801 cgt
//
