LOCUS       sequence-479075-        5294 bp    DNA     circular SYN 21-JUL-2025
DEFINITION  Express HiBiT-MRGPRX2 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 5294)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5294)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 21, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: HiBiT-MRGPRX2 Fusion Vector
FEATURES             Location/Qualifiers
     source          1..5294
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..721
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     672..692
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          857..989
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     972..991
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1033..1052
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1033..1051
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2237..2255)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2292..2413
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2303..2322)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2357..2376
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2658..3015
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2658..2849
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2681..2701)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2866..3001
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3030..3077
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3109..3903
                     /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(3163..3182)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3773..3792
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3967..4015
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(4254..4842)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4334..4353)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    4958..5241
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      661 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      721 tggtttagtg aaccgtcaga tcactagaag ctttattgcg gtagtttatc acagttaaat
      781 tgctaacgca gtcagtgctt ctgacacaac agtctcgaac ttaagctgca gaagttggtc
      841 gtgaggcact gggcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa
      901 actgggcttg tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac
      961 tgacatccac tttgcctttc tctccacagg tgtccactcc cagttcaatt acagctctta
     1021 aggctagagt attaatacga ctcactatag ggctagcgct caccatggtg agcggctggc
     1081 ggctgttcaa gaagattagc gggagctccg gtggctcgag cggtgcgatc gccatggatc
     1141 caaccacccc ggcctgggga acagaaagta caacagtgaa tggaaatgac caagcccttc
     1201 ttctgctttg tggcaaggag accctgatcc cggtcttcct gatccttttc attgccctgg
     1261 tcgggctggt aggaaacggg tttgtgctct ggctcctggg cttccgcatg cgcaggaacg
     1321 ccttctctgt ctacgtcctc agcctggccg gggccgactt cctcttcctc tgcttccaga
     1381 ttataaattg cctggtgtac ctcagtaact tcttctgttc catctccatc aatttcccta
     1441 gcttcttcac cactgtgatg acctgtgcct accttgcagg cctgagcatg ctgagcaccg
     1501 tcagcaccga gcgctgcctg tccgtcctgt ggcccatctg gtatcgctgc cgccgcccca
     1561 gacacctgtc agcggtcgtg tgtgtcctgc tctgggccct gtccctactg ctgagcatct
     1621 tggaagggaa gttctgtggc ttcttattta gtgatggtga ctctggttgg tgtcagacat
     1681 ttgatttcat cactgcagcg tggctgattt ttttattcat ggttctctgt gggtccagtc
     1741 tggccctgct ggtcaggatc ctctgtggct ccaggggtct gccactgacc aggctgtacc
     1801 tgaccatcct gctcacagtg ctggtgttcc tcctctgcgg cctgcccttt ggcattcagt
     1861 ggttcctaat attatggatc tggaaggatt ctgatgtctt attttgtcat attcatccag
     1921 tttcagttgt cctgtcatct cttaacagca gtgccaaccc catcatttac ttcttcgtgg
     1981 gctcttttag gaagcagtgg cggctgcagc agccgatcct caagctggct ctccagaggg
     2041 ctctgcagga cattgctgag gtggatcaca gtgaaggatg cttccgtcag ggcaccccgg
     2101 agatgtcgag aagcagtctg gtggtttaaa cgaattcggg ctcggtaccc ggggatcctc
     2161 tagagtcgac ctgcaggcat gcaagctgat ccggctgcta acaaagcccg aaaggaagct
     2221 gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc ggccgcttcg
     2281 agcagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     2341 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     2401 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggagat
     2461 gtgggaggtt tttttaagca agtaaaacct ctacaaatgt ggtaaaatcg aattttaaca
     2521 aaatattaac gcttacaatt tcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     2581 ttcacaccgc atacgcggat ctgcgcagca ccatggcctg aaataacctc tgaaagagga
     2641 acttggttag gtaccttctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg
     2701 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     2761 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     2821 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     2881 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     2941 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     3001 ctaggctttt gcaaaaagct taattaactg ttgacaatta atcatcggca tagtatgtcg
     3061 gcatagtata atacgacaag gtgaggaact aaacccagga ggcagatcat gattgaacaa
     3121 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     3181 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     3241 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     3301 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     3361 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     3421 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     3481 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     3541 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     3601 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     3661 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     3721 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     3781 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     3841 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     3901 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgat
     3961 ggccgcaata aaatatcttt attttcatta catctgtgtg ttggtttttt gtgtgaatcg
     4021 atagcgataa ggatcctctt tgcgcttgcg ttttcccttg tccagatagc ccagtagctg
     4081 acattcatcc ggggtcagca ccgtttctgc ggactggctt tctacccggt atcagctcac
     4141 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     4201 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     4261 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     4321 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     4381 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     4441 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     4501 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     4561 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     4621 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     4681 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     4741 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     4801 gtttgcaagc agcagattac gcgcagaaaa aaaggatttc aagaagatcc tttgatcttt
     4861 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     4921 ttatcaaaaa ggatcttcac ctagatcctt ttatagtccg gaaatacagg aacgcacgct
     4981 ggatggccct tcgctgggat ggtgaaacca tgaaaaatgg cagcttcagt ggattaagtg
     5041 ggggtaatgt ggcctgtacc ctctggttgc ataggtattc atacggttaa aatttatcag
     5101 gcgcgattgc ggcagttttt cgggtggttt gttgccattt ttacctgtct gctgccgtga
     5161 tcgcgctgaa cgcgttttag cggtgcgtac aattaaggga ttatggtaaa tccacttact
     5221 gtctgccctc gtagccatcg agataaaccg cagtactccg gccacgatgc gtccggcgta
     5281 gaggatcgag atct
//
