LOCUS       sequence-471922-        8754 bp    DNA     circular SYN 14-JUL-2025
DEFINITION  Express BCL9-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 8754)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 8754)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: BCL9-SmBiT Fusion Vector
FEATURES             Location/Qualifiers
     source          1..8754
                     /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    5643..5764
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5654..5673)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(5879..5912)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        5922..5969
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             6001..6795
                     /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(6055..6074)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6665..6684
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      7012..7557
                     /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     7458..7477
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7674..7691
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    7841..7889
                     /note="synthetic polyadenylation signal"
     CDS             complement(7913..8311)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      8308..8317
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(8342..8699)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8356..8491
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        8508..8699
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     8656..8676
                     /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 cgatcgccat gcattccagt
     1201 aaccctaaag tgaggagctc tccatcagga aacacacaga gtagccctaa gtcaaagcag
     1261 gaggtgatgg tccgtccccc tacagtgatg tccccatctg gaaaccccca gctggattcc
     1321 aaattctcca atcagggtaa acaggggggc tcagccagcc aatcccagcc atccccctgt
     1381 gactccaaga gtgggggcca tacccctaaa gcactccctg gcccaggtgg gagcatgggg
     1441 ctgaagaatg gggctggaaa tggtgccaag ggcaagggga aaagggagcg aagtatttcc
     1501 gccgactcct ttgatcagag agatcctggg actccaaacg atgactctga cattaaagaa
     1561 tgtaattctg ctgaccacat aaagtcccag gattcccagc acacaccaca ctcgatgacc
     1621 ccatcaaatg ctacagcccc caggtcttct accccctccc atggccaaac tactgccaca
     1681 gagcccacac ctgctcagaa gactccagcc aaagtggtgt acgtgttttc tactgagatg
     1741 gccaataaag ctgcagaagc tgttttgaag ggccaggttg aaactatcgt ctctttccac
     1801 atccagaaca tttctaacaa caagacagag agaagcacag cgcctctgaa cacacagata
     1861 tctgcccttc ggaatgatcc gaaacctctc ccacaacagc ccccagctcc ggccaaccag
     1921 gaccagaatt cttcccagaa taccagactg cagccaactc cacccattcc ggcaccagca
     1981 cccaagcctg ccgcaccccc acgtcccctg gaccgggaga gtcctggggt agaaaacaaa
     2041 ctgattcctt ctgtaggaag tcctgccagc tccactccac tgcccccaga tggtactggg
     2101 cccaactcaa ctcccaacaa tagggcagtg acccctgtct cccaggggag caatagctct
     2161 tcagcagatc ccaaagcccc tccgcctcca ccagtgtcca gtggcgagcc ccccacactg
     2221 ggagagaatc ccgatggcct atctcaggag cagctggagc accgggagcg ctccttacaa
     2281 actctcagag atatccagcg catgcttttt cctgatgaga aagaattcac aggagcacaa
     2341 agtgggggac cgcagcagaa tcctggggta ttagatgggc ctcagaaaaa accagaaggg
     2401 ccaatacagg ccatgatggc ccaatcccaa agcctaggta agggacctgg gccccggaca
     2461 gacgtgggag ctccatttgg ccctcaagga catagagatg tacccttttc tccagatgaa
     2521 atggttccac cttctatgaa ctcccagtct gggaccatag gacccgacca ccttgaccat
     2581 atgactcccg agcagatagc gtggctgaaa ctgcagcagg agttttatga agagaagagg
     2641 aggaagcagg aacaagtggt tgtccagcag tgttccctcc aggacatgat ggtccatcag
     2701 cacgggcctc ggggagtggt ccgaggaccc ccccctccat accagatgac ccctagtgaa
     2761 ggctgggcac ctgggggtac agagccattt tctgatggta tcaacatgcc acattctctg
     2821 cccccgaggg gcatggctcc ccaccccaac atgccaggga gccagatgcg cctccctgga
     2881 tttgcaggca tgataaactc tgaaatggaa gggccgaatg tccccaaccc tgcatctaga
     2941 ccaggtcttt ctggagtcag ttggccagat gatgtgccaa aaatcccaga tggtcgaaat
     3001 tttcctcctg gccagggcat tttcagcggt cctggccgag gggaacgctt cccaaacccc
     3061 caaggattgt ctgaagagat gtttcagcag cagctggcag agaaacagct gggtctcccc
     3121 ccagggatgg ccatggaagg catcaggccc agcatggaga tgaacaggat gattccaggc
     3181 tcccagcgcc acatggagcc tgggaataac cccattttcc ctcgaatacc agttgagggc
     3241 cctctgagtc cttctagggg tgactttcca aaaggaattc ccccacagat gggccctggt
     3301 cgggaacttg agtttgggat ggttcctagt gggatgaagg gagatgtcaa tctaaatgtc
     3361 aacatgggat ccaactctca gatgatacct cagaagatga gagaggctgg ggcgggccct
     3421 gaggagatgc tgaaattacg cccaggtggc tcagacatgc tgcctgctca gcagaagatg
     3481 gtgccactgc catttggtga gcacccccag caggagtatg gcatgggccc cagaccattc
     3541 cttcccatgt ctcagggtcc aggcagcaac agtggcttgc ggaatctcag agaaccaatt
     3601 gggcccgacc agaggactaa cagccggctc agtcatatgc caccactacc tctcaaccct
     3661 tccagtaacc ccaccagcct caacacagct cctccagttc agcgcggcct ggggcggaag
     3721 cccttggata tatctgtggc aggcagccag gtgcattccc caggcattaa ccctctgaag
     3781 tctcccacga tgcaccaagt ccagtcacca atgctgggct cgccctcggg gaacctcaag
     3841 tccccccaga ctccatcgca gctggcaggc atgctggcgg gcccagctgc tgctgcttcc
     3901 attaagtccc cccctgtttt ggggtctgct gctgcttcac ctgtccacct caagtctcca
     3961 tcacttcctg ccccgtcacc tggatggacc tcttctccaa aacctcccct tcagagtcct
     4021 gggatccctc caaaccataa agcacccctc accatggcct ccccagccat gctgggaaat
     4081 gtagagtcag gtggcccccc acctcctaca gccagccagc ctgcctctgt gaatatccct
     4141 ggaagtcttc cctctagtac accttatacc atgcctccag agccaaccct ttcccagaac
     4201 ccactctcta ttatgatgtc tcgaatgtcc aagtttgcaa tgcccagttc caccccgtta
     4261 taccatgatg ctatcaagac tgtggccagc tcagatgacg actcccctcc agctcgttct
     4321 cccaacttgc catcaatgaa taatatgcca ggaatgggca ttaatacaca gaatcctcga
     4381 atttcaggtc caaaccccgt ggttccgatg ccaaccctca gcccaatggg aatgacccag
     4441 ccactttctc actccaatca gatgccctct ccaaatgccg tgggacccaa catacctcct
     4501 catggggtcc caatggggcc tggcttgatg tcacacaatc ctatcatggg gcatgggtcc
     4561 caggagccac cgatggtacc tcaaggacgg atgggcttcc cccagggctt ccctccagta
     4621 cagtctcccc cacagcaggt tccattccct cacaatggcc ccagtggggg gcagggcagc
     4681 ttcccaggag ggatgggttt cccaggagaa ggcccccttg gccgccccag caacctgccc
     4741 caaagttcag cagatgcagc actttgcaag cctggaggcc ccgggggtcc tgactccttc
     4801 actgtcctgg ggaacagcat gccttcggtg tttacagacc cagatctgca ggaggtcatc
     4861 cgacctggag ccaccggaat acctgagttt gatctatccc gcattattcc atctgagaag
     4921 cccagccaga cgctgcaata tttccctcga ggggaagttc caggccgtaa acagccccag
     4981 ggtcctggac ctgggttttc acacatgcag gggatgatgg gcgaacaagc ccccagaatg
     5041 ggactagcat tacctggcat gggaggtcca gggccagtgg gaactccgga catccctctt
     5101 ggtacagctc catccatgcc aggccacaac cccatgagac caccagcctt tctccaacaa
     5161 ggcatgatgg gacctcacca tcggatgatg tcaccagcac aatctacaat gcccggccag
     5221 cccaccctga tgagcaatcc agctgctgcc gtgggcatga ttcctggcaa ggatcggggg
     5281 cctgccgggc tctacaccca ccctgggcct gtgggctctc caggcatgat gatgtccatg
     5341 cagggcatga tgggacccca acagaacatc atgatccccc cacagatgag gccccggggc
     5401 atggctgctg acgtgggcat gggtggattt agccaaggac ctggcaaccc aggaaacatg
     5461 atgtttgttt ctcaaggcag ttcaggtggt ggcgggagcg gaggtggagg ctcgagcggt
     5521 gtgaccggct accggctgtt cgaggagatt ctgtaaggcc gcgactctag agtcgacctg
     5581 caggcatgca agctgatttg gctgctgcca ccgctgagca aggccgcttc gagcagacat
     5641 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     5701 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     5761 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggaga tgtgggaggt
     5821 ttttttaagc aagtaaaacc tctacaaatg tggtaaaatc ttaattaaga attctaatat
     5881 aacttcgtat agcatacatt atacgaagtt atggatccac tgttgacaat taatcatcgg
     5941 catagtatat cggcatagta taatacgaca aggtgaggaa ctaaacccag gaggcagatc
     6001 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     6061 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     6121 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     6181 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     6241 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     6301 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     6361 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     6421 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     6481 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     6541 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     6601 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     6661 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     6721 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     6781 gacgagttct tctgattcga aatgaccgac caagcgacgc ccaaccggta tcagctcact
     6841 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtcaa
     6901 acatgagaat tacaacttat atcgtatggg gctgacttca ggtgctacat ttgaagagat
     6961 aaattgcact gaaatctaga aatattttat ctgattaata agatgatctt cttgagatcg
     7021 ttttggtctg cgcgtaatct cttgctctga aaacgaaaaa accgccttgc agggcggttt
     7081 ttcgaaggtt ctctgagcta ccaactcttt gaaccgaggt aactggcttg gaggagcgca
     7141 gtcaccaaaa cttgtccttt cagtttagcc ttaaccggcg catgacttca agactaactc
     7201 ctctaaatca attaccagtg gctgctgcca gtggtgcttt tgcatgtctt tccgggttgg
     7261 actcaagacg atagttaccg gataaggcgc agcggtcgga ctgaacgggg ggttcgtgca
     7321 tacagtccag cttggagcga actgcctacc cggaactgag tgtcaggcgt ggaatgagac
     7381 aaacgcggcc ataacagcgg aatgacaccg gtaaaccgaa aggcaggaac aggagagcgc
     7441 acgagggagc cgccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     7501 cactgatttg agcgtcagat ttcgtgatgc ttgtcagggg ggcggagcct atggaaaaac
     7561 ggctttgccg cggccctctc acttccctgt taagtatctt cctggcatct tccaggaaat
     7621 ctccgccccg ttcgtaagcc atttccgctc gccgcagtcg aacgaccgag cgtagcgagt
     7681 cagtgagcga ggaagcggaa tatatcctgt atcacatatt ctgctgacgc accggtgcag
     7741 ccttttttct cctgccacat gaagcacttc actgacaccc tcatcagtgc caacatagta
     7801 agccagtata cactccgcta gcgctgatgt ccggaatcgt cacacaaaaa accaacacac
     7861 agatgtaatg aaaataaaga tattttattg cggccatcgt gatggctaga gtttagccct
     7921 cccacacata accagagggc agcaattcac gaatcccaac tgccgtcggc tgtccatcac
     7981 tgtccttcac tatggctttg atcccaggat gcagatcgag aagcacctgt cggcaccgtc
     8041 cgcaggggct caagatgccc ctgttctcat ttccgatcgc gacgatacaa gtcaggttgc
     8101 cagctgccgc agcagcagca gtgcccagca ccacgagttc tgcacaaggt cccccagtaa
     8161 aatgatatac attgacacca gtgaagatgc ggccgtcgct agagagagct gcgctggcga
     8221 cgctgtagtc ttcagagatg gggatgctgt tgattgtagc cgttgctctt tcaatgaggg
     8281 tggattcttc ttgagacaaa ggcttggcca tggtggcgct agtgtcagaa gaatcgagct
     8341 ttttgcaaaa gcctaggcct ccaaaaaagc ctcctcacta cttctggaat agctcagagg
     8401 ccgaggcggc ctcggcctct gcataaataa aaaaaattag tcagccatgg ggcggagaat
     8461 gggcggaact gggcggagtt aggggcggga tgggcggagt taggggcggg actatggttg
     8521 ctgactaatt gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc
     8581 cacacctggt tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc
     8641 ctggggactt tccacaccct aactgacaca cattccacag ctggttcttt ccgcctcaga
     8701 aggtacctaa ccaagttcct ctttcagagg ttatttcagg ccatggtgct gcgc
//
