LOCUS       sequence-479841-        5766 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Express IFNGR1-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 5766)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5766)
  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            "Unpublished"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: IFNGR1-SmBiT Vector
FEATURES             Location/Qualifiers
     source          1..5766
                     /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(2709..2727)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2764..2885
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2775..2794)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2829..2848
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3130..3487
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3130..3321
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3153..3173)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3338..3473
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3502..3549
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3581..4375
                     /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(3635..3654)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4245..4264
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4439..4487
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(4726..5314)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4806..4825)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5430..5713
                     /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 cgccatggct ctcctctttc
     1081 tcctacccct tgtcatgcag ggtgtgagca gggctgagat gggcaccgcg gatctggggc
     1141 cgtcctcagt gcctacacca actaatgtta caattgaatc ctataacatg aaccctatcg
     1201 tatattggga gtaccagatc atgccacagg tccctgtttt taccgtagag gtaaagaact
     1261 atggtgttaa gaattcagaa tggattgatg cctgcatcaa tatttctcat cattattgta
     1321 atatttctga tcatgttggt gatccatcaa attctctttg ggtcagagtt aaagccaggg
     1381 ttggacaaaa agaatctgcc tatgcaaagt cagaagaatt tgctgtatgc cgagatggaa
     1441 aaattggacc acctaaactg gatatcagaa aggaggagaa gcaaatcatg attgacatat
     1501 ttcacccttc agtttttgta aatggagacg agcaggaagt cgattatgat cccgaaacta
     1561 cctgttacat tagggtgtac aatgtgtatg tgagaatgaa cggaagtgag atccagtata
     1621 aaatactcac gcagaaggaa gatgattgtg acgagattca gtgccagtta gcgattccag
     1681 tatcctcact gaattctcag tactgtgttt cagcagaagg agtcttacat gtgtggggtg
     1741 ttacaactga aaagtcaaaa gaagtttgta ttaccatttt caatagcagt ataaaaggtt
     1801 ctctttggat tccagttgtt gctgctttac tactctttct agtgcttagc ctggtattca
     1861 tctgttttta tattaagaaa attaatccat tgaaggaaaa aagcataata ttacccaagt
     1921 ccttgatctc tgtggtaaga agtgctactt tagagacaaa acctgaatca aaatatgtat
     1981 cactcatcac gtcataccag ccattttcct tagaaaagga ggtggtctgt gaagagccgt
     2041 tgtctccagc aacagttcca ggcatgcata ccgaagacaa tccaggaaaa gtggaacata
     2101 cagaagaact ttctagtata acagaagtgg tgactactga agaaaatatt cctgacgtgg
     2161 tcccgggcag ccatctgact ccaatagaga gagagagttc ttcaccttta agtagtaacc
     2221 agtctgaacc tggcagcatc gctttaaact cgtatcactc cagaaattgt tctgagagtg
     2281 atcactccag aaatggtttt gatactgatt ccagctgtct ggaatcacat agctccttat
     2341 ctgactcaga atttccccca aataataaag gtgaaataaa aacagaagga caagagctca
     2401 taaccgtaat aaaagccccc acctcctttg gttatgataa accacatgtg ctagtggatc
     2461 tacttgtgga tgatagcggt aaagagtcct tgattggtta tagaccaaca gaagattcca
     2521 aagaattttc agtttctcaa ggcagttcag gtggtggcgg gagcggaggt ggaggctcga
     2581 gcggtgtgac cggctaccgg ctgttcgagg agattctgta aggccgcgac tctagagtcg
     2641 acctgcaggc atgcaagctg atccggctgc taacaaagcc cgaaaggaag ctgagttggc
     2701 tgctgccacc gctgagcaat aactagcata accccttggg gcggccgctt cgagcagaca
     2761 tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct
     2821 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
     2881 aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag atgtgggagg
     2941 tttttttaag caagtaaaac ctctacaaat gtggtaaaat cgaattttaa caaaatatta
     3001 acgcttacaa tttcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     3061 gcatacgcgg atctgcgcag caccatggcc tgaaataacc tctgaaagag gaacttggtt
     3121 aggtaccttc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa
     3181 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3241 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     3301 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     3361 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     3421 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     3481 ttgcaaaaag cttaattaac tgttgacaat taatcatcgg catagtatat cggcatagta
     3541 taatacgaca aggtgaggaa ctaaacccag gaggcagatc atgattgaac aagatggatt
     3601 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     3661 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     3721 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct
     3781 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     3841 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     3901 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     3961 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     4021 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     4081 agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac
     4141 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     4201 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     4261 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     4321 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     4381 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg atggccgcaa
     4441 taaaatatct ttattttcat tacatctgtg tgttggtttt ttgtgtgaat cgatagcgat
     4501 aaggatcctc tttgcgcttg cgttttccct tgtccagata gcccagtagc tgacattcat
     4561 ccggggtcag caccgtttct gcggactggc tttctacccg gtatcagctc actcaaaggc
     4621 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     4681 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     4741 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     4801 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     4861 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     4921 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     4981 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     5041 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     5101 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     5161 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     5221 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     5281 gcagcagatt acgcgcagaa aaaaaggatt tcaagaagat cctttgatct tttctacggg
     5341 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     5401 aaggatcttc acctagatcc ttttatagtc cggaaataca ggaacgcacg ctggatggcc
     5461 cttcgctggg atggtgaaac catgaaaaat ggcagcttca gtggattaag tgggggtaat
     5521 gtggcctgta ccctctggtt gcataggtat tcatacggtt aaaatttatc aggcgcgatt
     5581 gcggcagttt ttcgggtggt ttgttgccat ttttacctgt ctgctgccgt gatcgcgctg
     5641 aacgcgtttt agcggtgcgt acaattaagg gattatggta aatccactta ctgtctgccc
     5701 tcgtagccat cgagataaac cgcagtactc cggccacgat gcgtccggcg tagaggatcg
     5761 agatct
//
