LOCUS       sequence-472439-        8813 bp    DNA     circular SYN 13-MAY-2025
DEFINITION  Express SmBiT-BCL9 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 8813)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 8813)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: SmBiT-BCL9 Fusion Vector
FEATURES             Location/Qualifiers
     source          1..8813
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    24..105
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     enhancer        338..641
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        642..845
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     796..816
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          981..1113
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1096..1115
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1157..1176
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1157..1175
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    5702..5823
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5713..5732)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(5938..5971)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        5981..6028
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             6060..6854
                     /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="MIEQDGLQAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6114..6133)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6724..6743
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      7071..7616
                     /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     7517..7536
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7733..7750
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    7900..7948
                     /note="synthetic polyadenylation signal"
     CDS             complement(7972..8370)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      8367..8376
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(8401..8758)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8415..8550
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        8567..8758
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     8715..8735
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 aagacgcgtt aattaagacg tcggcagtga aaaaaatgct ttatttgtga aatttgtgat
       61 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
      121 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac
      181 ctctacaaat gtggtatggc tgattatgat ccgcggccgc aatacgtcga cgttatcagc
      241 tgacttcgta cgagagccta ggattatggc gcgccactag ttattaatag taatcaatta
      301 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      361 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      421 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      481 ctgcccactt ggcagtacat caagtgtatc atatgccaag tccgccccct attgacgtca
      541 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttacgg gactttccta
      601 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      661 acaccaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      721 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaata
      781 accccgcccc gttgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      841 gagctggttt agtgaaccgt cagatcacta gaagctttat tgcggtagtt tatcacagtt
      901 aaattgctaa cgcagtcagt gcttctgaca caacagtctc gaacttaagc tgcagaagtt
      961 ggtcgtgagg cactgggcag gtaagtatca aggttacaag acaggtttaa ggagaccaat
     1021 agaaactggg cttgtcgaga cagagaagac tcttgcgttt ctgataggca cctattggtc
     1081 ttactgacat ccactttgcc tttctctcca caggtgtcca ctcccagttc aattacagct
     1141 cttaaggcta gagtattaat acgactcact atagggctag cgctcaccat ggtgaccggc
     1201 taccggctgt tcgaggagat tctcgggagc tccggtggtg gcgggagcgg aggtggaggc
     1261 tcgagcggtg cgatcgccat gcattccagt aaccctaaag tgaggagctc tccatcagga
     1321 aacacacaga gtagccctaa gtcaaagcag gaggtgatgg tccgtccccc tacagtgatg
     1381 tccccatctg gaaaccccca gctggattcc aaattctcca atcagggtaa acaggggggc
     1441 tcagccagcc aatcccagcc atccccctgt gactccaaga gtgggggcca tacccctaaa
     1501 gcactccctg gcccaggtgg gagcatgggg ctgaagaatg gggctggaaa tggtgccaag
     1561 ggcaagggga aaagggagcg aagtatttcc gccgactcct ttgatcagag agatcctggg
     1621 actccaaacg atgactctga cattaaagaa tgtaattctg ctgaccacat aaagtcccag
     1681 gattcccagc acacaccaca ctcgatgacc ccatcaaatg ctacagcccc caggtcttct
     1741 accccctccc atggccaaac tactgccaca gagcccacac ctgctcagaa gactccagcc
     1801 aaagtggtgt acgtgttttc tactgagatg gccaataaag ctgcagaagc tgttttgaag
     1861 ggccaggttg aaactatcgt ctctttccac atccagaaca tttctaacaa caagacagag
     1921 agaagcacag cgcctctgaa cacacagata tctgcccttc ggaatgatcc gaaacctctc
     1981 ccacaacagc ccccagctcc ggccaaccag gaccagaatt cttcccagaa taccagactg
     2041 cagccaactc cacccattcc ggcaccagca cccaagcctg ccgcaccccc acgtcccctg
     2101 gaccgggaga gtcctggggt agaaaacaaa ctgattcctt ctgtaggaag tcctgccagc
     2161 tccactccac tgcccccaga tggtactggg cccaactcaa ctcccaacaa tagggcagtg
     2221 acccctgtct cccaggggag caatagctct tcagcagatc ccaaagcccc tccgcctcca
     2281 ccagtgtcca gtggcgagcc ccccacactg ggagagaatc ccgatggcct atctcaggag
     2341 cagctggagc accgggagcg ctccttacaa actctcagag atatccagcg catgcttttt
     2401 cctgatgaga aagaattcac aggagcacaa agtgggggac cgcagcagaa tcctggggta
     2461 ttagatgggc ctcagaaaaa accagaaggg ccaatacagg ccatgatggc ccaatcccaa
     2521 agcctaggta agggacctgg gccccggaca gacgtgggag ctccatttgg ccctcaagga
     2581 catagagatg tacccttttc tccagatgaa atggttccac cttctatgaa ctcccagtct
     2641 gggaccatag gacccgacca ccttgaccat atgactcccg agcagatagc gtggctgaaa
     2701 ctgcagcagg agttttatga agagaagagg aggaagcagg aacaagtggt tgtccagcag
     2761 tgttccctcc aggacatgat ggtccatcag cacgggcctc ggggagtggt ccgaggaccc
     2821 ccccctccat accagatgac ccctagtgaa ggctgggcac ctgggggtac agagccattt
     2881 tctgatggta tcaacatgcc acattctctg cccccgaggg gcatggctcc ccaccccaac
     2941 atgccaggga gccagatgcg cctccctgga tttgcaggca tgataaactc tgaaatggaa
     3001 gggccgaatg tccccaaccc tgcatctaga ccaggtcttt ctggagtcag ttggccagat
     3061 gatgtgccaa aaatcccaga tggtcgaaat tttcctcctg gccagggcat tttcagcggt
     3121 cctggccgag gggaacgctt cccaaacccc caaggattgt ctgaagagat gtttcagcag
     3181 cagctggcag agaaacagct gggtctcccc ccagggatgg ccatggaagg catcaggccc
     3241 agcatggaga tgaacaggat gattccaggc tcccagcgcc acatggagcc tgggaataac
     3301 cccattttcc ctcgaatacc agttgagggc cctctgagtc cttctagggg tgactttcca
     3361 aaaggaattc ccccacagat gggccctggt cgggaacttg agtttgggat ggttcctagt
     3421 gggatgaagg gagatgtcaa tctaaatgtc aacatgggat ccaactctca gatgatacct
     3481 cagaagatga gagaggctgg ggcgggccct gaggagatgc tgaaattacg cccaggtggc
     3541 tcagacatgc tgcctgctca gcagaagatg gtgccactgc catttggtga gcacccccag
     3601 caggagtatg gcatgggccc cagaccattc cttcccatgt ctcagggtcc aggcagcaac
     3661 agtggcttgc ggaatctcag agaaccaatt gggcccgacc agaggactaa cagccggctc
     3721 agtcatatgc caccactacc tctcaaccct tccagtaacc ccaccagcct caacacagct
     3781 cctccagttc agcgcggcct ggggcggaag cccttggata tatctgtggc aggcagccag
     3841 gtgcattccc caggcattaa ccctctgaag tctcccacga tgcaccaagt ccagtcacca
     3901 atgctgggct cgccctcggg gaacctcaag tccccccaga ctccatcgca gctggcaggc
     3961 atgctggcgg gcccagctgc tgctgcttcc attaagtccc cccctgtttt ggggtctgct
     4021 gctgcttcac ctgtccacct caagtctcca tcacttcctg ccccgtcacc tggatggacc
     4081 tcttctccaa aacctcccct tcagagtcct gggatccctc caaaccataa agcacccctc
     4141 accatggcct ccccagccat gctgggaaat gtagagtcag gtggcccccc acctcctaca
     4201 gccagccagc ctgcctctgt gaatatccct ggaagtcttc cctctagtac accttatacc
     4261 atgcctccag agccaaccct ttcccagaac ccactctcta ttatgatgtc tcgaatgtcc
     4321 aagtttgcaa tgcccagttc caccccgtta taccatgatg ctatcaagac tgtggccagc
     4381 tcagatgacg actcccctcc agctcgttct cccaacttgc catcaatgaa taatatgcca
     4441 ggaatgggca ttaatacaca gaatcctcga atttcaggtc caaaccccgt ggttccgatg
     4501 ccaaccctca gcccaatggg aatgacccag ccactttctc actccaatca gatgccctct
     4561 ccaaatgccg tgggacccaa catacctcct catggggtcc caatggggcc tggcttgatg
     4621 tcacacaatc ctatcatggg gcatgggtcc caggagccac cgatggtacc tcaaggacgg
     4681 atgggcttcc cccagggctt ccctccagta cagtctcccc cacagcaggt tccattccct
     4741 cacaatggcc ccagtggggg gcagggcagc ttcccaggag ggatgggttt cccaggagaa
     4801 ggcccccttg gccgccccag caacctgccc caaagttcag cagatgcagc actttgcaag
     4861 cctggaggcc ccgggggtcc tgactccttc actgtcctgg ggaacagcat gccttcggtg
     4921 tttacagacc cagatctgca ggaggtcatc cgacctggag ccaccggaat acctgagttt
     4981 gatctatccc gcattattcc atctgagaag cccagccaga cgctgcaata tttccctcga
     5041 ggggaagttc caggccgtaa acagccccag ggtcctggac ctgggttttc acacatgcag
     5101 gggatgatgg gcgaacaagc ccccagaatg ggactagcat tacctggcat gggaggtcca
     5161 gggccagtgg gaactccgga catccctctt ggtacagctc catccatgcc aggccacaac
     5221 cccatgagac caccagcctt tctccaacaa ggcatgatgg gacctcacca tcggatgatg
     5281 tcaccagcac aatctacaat gcccggccag cccaccctga tgagcaatcc agctgctgcc
     5341 gtgggcatga ttcctggcaa ggatcggggg cctgccgggc tctacaccca ccctgggcct
     5401 gtgggctctc caggcatgat gatgtccatg cagggcatga tgggacccca acagaacatc
     5461 atgatccccc cacagatgag gccccggggc atggctgctg acgtgggcat gggtggattt
     5521 agccaaggac ctggcaaccc aggaaacatg atgtttgttt aaacgaattc gggctcggta
     5581 cccggggatc ctctagagtc gacctgcagg catgcaagct gatccggctg ctaacaaagc
     5641 ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa ggccgcttcg agcagacatg
     5701 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
     5761 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
     5821 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggagat gtgggaggtt
     5881 tttttaagca agtaaaacct ctacaaatgt ggtaaaatct taattaagaa ttctaatata
     5941 acttcgtata gcatacatta tacgaagtta tggatccact gttgacaatt aatcatcggc
     6001 atagtatatc ggcatagtat aatacgacaa ggtgaggaac taaacccagg aggcagatca
     6061 tgattgaaca agatggattg caggcaggtt ctccggccgc ttgggtggag aggctattcg
     6121 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     6181 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     6241 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     6301 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     6361 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     6421 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     6481 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     6541 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
     6601 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     6661 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     6721 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     6781 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     6841 acgagttctt ctgattcgaa atgaccgacc aagcgacgcc caaccggtat cagctcactc
     6901 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtcaaa
     6961 catgagaatt acaacttata tcgtatgggg ctgacttcag gtgctacatt tgaagagata
     7021 aattgcactg aaatctagaa atattttatc tgattaataa gatgatcttc ttgagatcgt
     7081 tttggtctgc gcgtaatctc ttgctctgaa aacgaaaaaa ccgccttgca gggcggtttt
     7141 tcgaaggttc tctgagctac caactctttg aaccgaggta actggcttgg aggagcgcag
     7201 tcaccaaaac ttgtcctttc agtttagcct taaccggcgc atgacttcaa gactaactcc
     7261 tctaaatcaa ttaccagtgg ctgctgccag tggtgctttt gcatgtcttt ccgggttgga
     7321 ctcaagacga tagttaccgg ataaggcgca gcggtcggac tgaacggggg gttcgtgcat
     7381 acagtccagc ttggagcgaa ctgcctaccc ggaactgagt gtcaggcgtg gaatgagaca
     7441 aacgcggcca taacagcgga atgacaccgg taaaccgaaa ggcaggaaca ggagagcgca
     7501 cgagggagcc gccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     7561 actgatttga gcgtcagatt tcgtgatgct tgtcaggggg gcggagccta tggaaaaacg
     7621 gctttgccgc ggccctctca cttccctgtt aagtatcttc ctggcatctt ccaggaaatc
     7681 tccgccccgt tcgtaagcca tttccgctcg ccgcagtcga acgaccgagc gtagcgagtc
     7741 agtgagcgag gaagcggaat atatcctgta tcacatattc tgctgacgca ccggtgcagc
     7801 cttttttctc ctgccacatg aagcacttca ctgacaccct catcagtgcc aacatagtaa
     7861 gccagtatac actccgctag cgctgatgtc cggaatcgtc acacaaaaaa ccaacacaca
     7921 gatgtaatga aaataaagat attttattgc ggccatcgtg atggctagag tttagccctc
     7981 ccacacataa ccagagggca gcaattcacg aatcccaact gccgtcggct gtccatcact
     8041 gtccttcact atggctttga tcccaggatg cagatcgaga agcacctgtc ggcaccgtcc
     8101 gcaggggctc aagatgcccc tgttctcatt tccgatcgcg acgatacaag tcaggttgcc
     8161 agctgccgca gcagcagcag tgcccagcac cacgagttct gcacaaggtc ccccagtaaa
     8221 atgatataca ttgacaccag tgaagatgcg gccgtcgcta gagagagctg cgctggcgac
     8281 gctgtagtct tcagagatgg ggatgctgtt gattgtagcc gttgctcttt caatgagggt
     8341 ggattcttct tgagacaaag gcttggccat ggtggcgcta gtgtcagaag aatcgagctt
     8401 tttgcaaaag cctaggcctc caaaaaagcc tcctcactac ttctggaata gctcagaggc
     8461 cgaggcggcc tcggcctctg cataaataaa aaaaattagt cagccatggg gcggagaatg
     8521 ggcggaactg ggcggagtta ggggcgggat gggcggagtt aggggcggga ctatggttgc
     8581 tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc
     8641 acacctggtt gctgactaat tgagatgcat gctttgcata cttctgcctg ctggggagcc
     8701 tggggacttt ccacacccta actgacacac attccacagc tggttctttc cgcctcagaa
     8761 ggtacctaac caagttcctc tttcagaggt tatttcaggc catggtgctg cgc
//
