LOCUS       sequence-478477-        6090 bp    DNA     circular SYN 05-AUG-2025
DEFINITION  Express BIRC3-HiBiT 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 6090)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6090)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 5, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: BIRC3-HiBiT Fusion Vector
FEATURES             Location/Qualifiers
     source          1..6090
                     /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(3033..3051)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3088..3209
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3099..3118)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3153..3172
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3454..3811
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3454..3645
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3477..3497)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3662..3797
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3826..3873
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3905..4699
                     /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(3959..3978)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4569..4588
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4763..4811
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(5050..5638)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5130..5149)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5754..6037
                     /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 cgccatgaac atagtagaaa
     1081 acagcatatt cttatcaaat ttgatgaaaa gcgccaacac gtttgaactg aaatacgact
     1141 tgtcatgtga actgtaccga atgtctacgt attccacttt tcctgctggg gttcctgtct
     1201 cagaaaggag tcttgctcgt gctggtttct attacactgg tgtgaatgac aaggtcaaat
     1261 gcttctgttg tggcctgatg ctggataact ggaaaagagg agacagtcct actgaaaagc
     1321 ataaaaagtt gtatcctagc tgcagattcg ttcagagtct aaattccgtt aacaacttgg
     1381 aagctacctc tcagcctact tttccttctt cagtaacaaa ttccacacac tcattacttc
     1441 cgggtacaga aaacagtgga tatttccgtg gctcttattc aaactctcca tcaaatcctg
     1501 taaactccag agcaaatcaa gatttttctg ccttgatgag aagttcctac cactgtgcaa
     1561 tgaataacga aaatgccaga ttacttactt ttcagacatg gccattgact tttctgtcgc
     1621 caacagatct ggcaaaagca ggcttttact acataggacc tggagacaga gtggcttgct
     1681 ttgcctgtgg tggaaaattg agcaattggg aaccgaagga taatgctatg tcagaacacc
     1741 tgagacattt tcccaaatgc ccatttatag aaaatcagct tcaagacact tcaagataca
     1801 cagtttctaa tctgagcatg cagacacatg cagcccgctt taaaacattc tttaactggc
     1861 cctctagtgt tctagttaat cctgagcagc ttgcaagtgc gggtttttat tatgtgggta
     1921 acagtgatga tgtcaaatgc ttttgctgtg atggtggact caggtgttgg gaatctggag
     1981 atgatccatg ggttcaacat gccaagtggt ttccaaggtg tgagtacttg ataagaatta
     2041 aaggacagga gttcatccgt caagttcaag ccagttaccc tcatctactt gaacagctgc
     2101 tatccacatc agacagccca ggagatgaaa atgcagagtc atcaattatc cattttgaac
     2161 ctggagaaga ccattcagaa gatgcaatca tgatgaatac tcctgtgatt aatgctgccg
     2221 tggaaatggg ctttagtaga agcctggtaa aacagacagt tcagagaaaa atcctagcaa
     2281 ctggagagaa ttatagacta gtcaatgatc ttgtgttaga cttactcaat gcagaagatg
     2341 aaataaggga agaggagaga gaaagagcaa ctgaggaaaa agaatcaaat gatttattat
     2401 taatccggaa gaatagaatg gcactttttc aacatttgac ttgtgtaatt ccaatcctgg
     2461 atagtctact aactgccgga attattaatg aacaagaaca tgatgttatt aaacagaaga
     2521 cacagacgtc tttacaagca agagaactga ttgatacgat tttagtaaaa ggaaatattg
     2581 cagccactgt attcagaaac tctctgcaag aagctgaagc tgtgttatat gagcatttat
     2641 ttgtgcaaca ggacataaaa tatattccca cagaagatgt ttcagatcta ccagtggaag
     2701 aacaattgcg gagactacaa gaagaaagaa catgtaaagt gtgtatggac aaagaagtgt
     2761 ccatagtgtt tattccttgt ggtcatctag tagtatgcaa agattgtgct ccttctttaa
     2821 gaaagtgtcc tatttgtagg agtacaatca agggtacagt tcgtacattt ctttcagttt
     2881 ctcaaggcag ttcaggtggc tcgagcggtg tgagcggctg gcggctgttc aagaagatta
     2941 gctaaggccg cgactctaga gtcgacctgc aggcatgcaa gctgatccgg ctgctaacaa
     3001 agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactag cataacccct
     3061 tggggcggcc gcttcgagca gacatgataa gatacattga tgagtttgga caaaccacaa
     3121 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
     3181 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
     3241 aggttcaggg ggagatgtgg gaggtttttt taagcaagta aaacctctac aaatgtggta
     3301 aaatcgaatt ttaacaaaat attaacgctt acaatttcct gatgcggtat tttctcctta
     3361 cgcatctgtg cggtatttca caccgcatac gcggatctgc gcagcaccat ggcctgaaat
     3421 aacctctgaa agaggaactt ggttaggtac cttctgaggc ggaaagaacc agctgtggaa
     3481 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3541 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     3601 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     3661 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     3721 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     3781 aggctttttt ggaggcctag gcttttgcaa aaagcttaat taactgttga caattaatca
     3841 tcggcatagt atatcggcat agtataatac gacaaggtga ggaactaaac ccaggaggca
     3901 gatcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     3961 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     4021 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     4081 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     4141 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     4201 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     4261 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     4321 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     4381 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc
     4441 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     4501 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     4561 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     4621 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     4681 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     4741 aacctgccat cacgatggcc gcaataaaat atctttattt tcattacatc tgtgtgttgg
     4801 ttttttgtgt gaatcgatag cgataaggat cctctttgcg cttgcgtttt cccttgtcca
     4861 gatagcccag tagctgacat tcatccgggg tcagcaccgt ttctgcggac tggctttcta
     4921 cccggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     4981 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     5041 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     5101 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     5161 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     5221 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     5281 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     5341 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     5401 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     5461 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     5521 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     5581 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatttcaaga
     5641 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     5701 gattttggtc atgagattat caaaaaggat cttcacctag atccttttat agtccggaaa
     5761 tacaggaacg cacgctggat ggcccttcgc tgggatggtg aaaccatgaa aaatggcagc
     5821 ttcagtggat taagtggggg taatgtggcc tgtaccctct ggttgcatag gtattcatac
     5881 ggttaaaatt tatcaggcgc gattgcggca gtttttcggg tggtttgttg ccatttttac
     5941 ctgtctgctg ccgtgatcgc gctgaacgcg ttttagcggt gcgtacaatt aagggattat
     6001 ggtaaatcca cttactgtct gccctcgtag ccatcgagat aaaccgcagt actccggcca
     6061 cgatgcgtcc ggcgtagagg atcgagatct
//
