LOCUS       sequence-479840-        5310 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Express IFNGR2-SmBiT 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 5310)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5310)
  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            "Unpublished"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: IFNGR2-SmBiT Vector
FEATURES             Location/Qualifiers
     source          1..5310
                     /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(2253..2271)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2308..2429
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2319..2338)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2373..2392
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2674..3031
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2674..2865
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2697..2717)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2882..3017
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3046..3093
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3125..3919
                     /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(3179..3198)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3789..3808
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3983..4031
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(4270..4858)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4350..4369)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    4974..5257
                     /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 ggctagcgat cgccatgcga ccgacgctgc
     1081 tgtggtcgct gctgctgctg ctcggagtct tcgccgccgc cgccgcggcc ccgccagacc
     1141 ctctttccca gctgcccgct cctcagcacc cgaagattcg cctgtacaac gcagagcagg
     1201 tcctgagttg ggagccagtg gccctgagca atagcacgag gcctgttgtc taccaagtgc
     1261 agtttaaata caccgacagt aaatggttca cggccgacat catgtccata ggggtgaatt
     1321 gtacacagat cacagcaaca gagtgtgact tcactgccgc cagtccctca gcaggcttcc
     1381 caatggattt caatgtcact ctacgccttc gagctgagct gggagcactc cattctgcct
     1441 gggtgacaat gccttggttt caacactatc ggaatgtgac tgtcgggcct ccagaaaaca
     1501 ttgaggtgac cccaggagaa ggctccctca tcatcaggtt ctcctctccc tttgacatcg
     1561 ctgatacctc cacggccttt ttttgttatt atgtccatta ctgggaaaaa ggaggaatcc
     1621 aacaggtcaa aggccctttc agaagcaact ccatttcatt ggataactta aaaccctcca
     1681 gagtgtactg tttacaagtc caggcacaac tgctttggaa caaaagtaac atctttagag
     1741 tcgggcattt aagcaacata tcttgctacg aaacaatggc agatgcctcc actgagcttc
     1801 agcaagtcat cctgatctcc gtgggaacat tttcgttgct gtcggtgctg gcaggagcct
     1861 gtttcttcct ggtcctgaaa tatagaggcc tgattaaata ctggtttcac actccaccaa
     1921 gcatcccatt acagatagaa gagtatttaa aagacccaac tcagcccatc ttagaggcct
     1981 tggacaagga cagctcacca aaggatgacg tctgggactc tgtgtccatt atctcgtttc
     2041 cggaaaagga gcaagaagat gttctccaaa cgcttgtttc tcaaggcagt tcaggtggtg
     2101 gcgggagcgg aggtggaggc tcgagcggtg tgaccggcta ccggctgttc gaggagattc
     2161 tgtaaggccg cgactctaga gtcgacctgc aggcatgcaa gctgatccgg ctgctaacaa
     2221 agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactag cataacccct
     2281 tggggcggcc gcttcgagca gacatgataa gatacattga tgagtttgga caaaccacaa
     2341 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
     2401 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
     2461 aggttcaggg ggagatgtgg gaggtttttt taagcaagta aaacctctac aaatgtggta
     2521 aaatcgaatt ttaacaaaat attaacgctt acaatttcct gatgcggtat tttctcctta
     2581 cgcatctgtg cggtatttca caccgcatac gcggatctgc gcagcaccat ggcctgaaat
     2641 aacctctgaa agaggaactt ggttaggtac cttctgaggc ggaaagaacc agctgtggaa
     2701 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     2761 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     2821 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     2881 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     2941 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     3001 aggctttttt ggaggcctag gcttttgcaa aaagcttaat taactgttga caattaatca
     3061 tcggcatagt atatcggcat agtataatac gacaaggtga ggaactaaac ccaggaggca
     3121 gatcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     3181 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     3241 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     3301 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     3361 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     3421 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     3481 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     3541 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     3601 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc
     3661 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     3721 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     3781 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     3841 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     3901 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     3961 aacctgccat cacgatggcc gcaataaaat atctttattt tcattacatc tgtgtgttgg
     4021 ttttttgtgt gaatcgatag cgataaggat cctctttgcg cttgcgtttt cccttgtcca
     4081 gatagcccag tagctgacat tcatccgggg tcagcaccgt ttctgcggac tggctttcta
     4141 cccggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     4201 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     4261 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     4321 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     4381 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     4441 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     4501 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     4561 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     4621 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     4681 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     4741 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     4801 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatttcaaga
     4861 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     4921 gattttggtc atgagattat caaaaaggat cttcacctag atccttttat agtccggaaa
     4981 tacaggaacg cacgctggat ggcccttcgc tgggatggtg aaaccatgaa aaatggcagc
     5041 ttcagtggat taagtggggg taatgtggcc tgtaccctct ggttgcatag gtattcatac
     5101 ggttaaaatt tatcaggcgc gattgcggca gtttttcggg tggtttgttg ccatttttac
     5161 ctgtctgctg ccgtgatcgc gctgaacgcg ttttagcggt gcgtacaatt aagggattat
     5221 ggtaaatcca cttactgtct gccctcgtag ccatcgagat aaaccgcagt actccggcca
     5281 cgatgcgtcc ggcgtagagg atcgagatct
//
