LOCUS       sequence-471921-        9242 bp    DNA     circular SYN 14-JUL-2025
DEFINITION  Express BCL9-LgBiT 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 9242)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 9242)
  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-LgBiT Fusion Vector
FEATURES             Location/Qualifiers
     source          1..9242
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    17..98
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     42..61
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(5122..5140)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(5123..5140)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     enhancer        5260..5563
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     5718..5738
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        5771..5818
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5850..6644
                     /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(5904..5923)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6514..6533
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      6861..7406
                     /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     7307..7326
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7523..7540
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    7690..7738
                     /note="synthetic polyadenylation signal"
     CDS             complement(7762..8799)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             7775..7783
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     promoter        complement(8830..9187)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8844..8979
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        8996..9187
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     9144..9164
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 aagacgcgtg acgtcggcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
       61 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
      121 ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca
      181 aatgtggtat ggctgattat gatccgcggc cgcaatacgt cgacgttatc agctgacttc
      241 gtacgagagc ctaggtgttt ctagagtcgc ggccttagct gttgatggtt actcggaaca
      301 gcatggagcc gtcgggggtg atcaggcgct cgtcgataat tttgttgccg ttccacaggg
      361 tccctgttac agtgatcttt ttgccgtcga acacggcgat gccttcatac ggccgtccga
      421 aatagttcag catgttcggc gtaaccccgt cgattaccag tgtgccatag ggcaggatca
      481 ccttaaagtg atgatcatcc acagggtaca ccaccttaaa cacctcttcg atctgggcca
      541 tttggtcggc gctcagacct tcatacggga tgatgacatg gatgtcgatc ttcagggcat
      601 tttcaccgct ccggacaatc ctttggatcg gagttacgga cacggcgaga ttctgcagca
      661 aactggacac acctccctgt tcaaggactt ggtccaggtt gtaggcggct gtctgttccc
      721 agtccccaac gaaatcttcg agtgtgaaga caccgctcga gcctccacct ccgctcccgc
      781 caccacctga actgccttga gaaacaaaca tcatgtttcc tgggttgcca ggtccttggc
      841 taaatccacc catgcccacg tcagcagcca tgccccgggg cctcatctgt ggggggatca
      901 tgatgttctg ttggggtccc atcatgccct gcatggacat catcatgcct ggagagccca
      961 caggcccagg gtgggtgtag agcccggcag gcccccgatc cttgccagga atcatgccca
     1021 cggcagcagc tggattgctc atcagggtgg gctggccggg cattgtagat tgtgctggtg
     1081 acatcatccg atggtgaggt cccatcatgc cttgttggag aaaggctggt ggtctcatgg
     1141 ggttgtggcc tggcatggat ggagctgtac caagagggat gtccggagtt cccactggcc
     1201 ctggacctcc catgccaggt aatgctagtc ccattctggg ggcttgttcg cccatcatcc
     1261 cctgcatgtg tgaaaaccca ggtccaggac cctggggctg tttacggcct ggaacttccc
     1321 ctcgagggaa atattgcagc gtctggctgg gcttctcaga tggaataatg cgggatagat
     1381 caaactcagg tattccggtg gctccaggtc ggatgacctc ctgcagatct gggtctgtaa
     1441 acaccgaagg catgctgttc cccaggacag tgaaggagtc aggacccccg gggcctccag
     1501 gcttgcaaag tgctgcatct gctgaacttt ggggcaggtt gctggggcgg ccaagggggc
     1561 cttctcctgg gaaacccatc cctcctggga agctgccctg ccccccactg gggccattgt
     1621 gagggaatgg aacctgctgt gggggagact gtactggagg gaagccctgg gggaagccca
     1681 tccgtccttg aggtaccatc ggtggctcct gggacccatg ccccatgata ggattgtgtg
     1741 acatcaagcc aggccccatt gggaccccat gaggaggtat gttgggtccc acggcatttg
     1801 gagagggcat ctgattggag tgagaaagtg gctgggtcat tcccattggg ctgagggttg
     1861 gcatcggaac cacggggttt ggacctgaaa ttcgaggatt ctgtgtatta atgcccattc
     1921 ctggcatatt attcattgat ggcaagttgg gagaacgagc tggaggggag tcgtcatctg
     1981 agctggccac agtcttgata gcatcatggt ataacggggt ggaactgggc attgcaaact
     2041 tggacattcg agacatcata atagagagtg ggttctggga aagggttggc tctggaggca
     2101 tggtataagg tgtactagag ggaagacttc cagggatatt cacagaggca ggctggctgg
     2161 ctgtaggagg tggggggcca cctgactcta catttcccag catggctggg gaggccatgg
     2221 tgaggggtgc tttatggttt ggagggatcc caggactctg aaggggaggt tttggagaag
     2281 aggtccatcc aggtgacggg gcaggaagtg atggagactt gaggtggaca ggtgaagcag
     2341 cagcagaccc caaaacaggg ggggacttaa tggaagcagc agcagctggg cccgccagca
     2401 tgcctgccag ctgcgatgga gtctgggggg acttgaggtt ccccgagggc gagcccagca
     2461 ttggtgactg gacttggtgc atcgtgggag acttcagagg gttaatgcct ggggaatgca
     2521 cctggctgcc tgccacagat atatccaagg gcttccgccc caggccgcgc tgaactggag
     2581 gagctgtgtt gaggctggtg gggttactgg aagggttgag aggtagtggt ggcatatgac
     2641 tgagccggct gttagtcctc tggtcgggcc caattggttc tctgagattc cgcaagccac
     2701 tgttgctgcc tggaccctga gacatgggaa ggaatggtct ggggcccatg ccatactcct
     2761 gctgggggtg ctcaccaaat ggcagtggca ccatcttctg ctgagcaggc agcatgtctg
     2821 agccacctgg gcgtaatttc agcatctcct cagggcccgc cccagcctct ctcatcttct
     2881 gaggtatcat ctgagagttg gatcccatgt tgacatttag attgacatct cccttcatcc
     2941 cactaggaac catcccaaac tcaagttccc gaccagggcc catctgtggg ggaattcctt
     3001 ttggaaagtc acccctagaa ggactcagag ggccctcaac tggtattcga gggaaaatgg
     3061 ggttattccc aggctccatg tggcgctggg agcctggaat catcctgttc atctccatgc
     3121 tgggcctgat gccttccatg gccatccctg gggggagacc cagctgtttc tctgccagct
     3181 gctgctgaaa catctcttca gacaatcctt gggggtttgg gaagcgttcc cctcggccag
     3241 gaccgctgaa aatgccctgg ccaggaggaa aatttcgacc atctgggatt tttggcacat
     3301 catctggcca actgactcca gaaagacctg gtctagatgc agggttgggg acattcggcc
     3361 cttccatttc agagtttatc atgcctgcaa atccagggag gcgcatctgg ctccctggca
     3421 tgttggggtg gggagccatg cccctcgggg gcagagaatg tggcatgttg ataccatcag
     3481 aaaatggctc tgtaccccca ggtgcccagc cttcactagg ggtcatctgg tatggagggg
     3541 ggggtcctcg gaccactccc cgaggcccgt gctgatggac catcatgtcc tggagggaac
     3601 actgctggac aaccacttgt tcctgcttcc tcctcttctc ttcataaaac tcctgctgca
     3661 gtttcagcca cgctatctgc tcgggagtca tatggtcaag gtggtcgggt cctatggtcc
     3721 cagactggga gttcatagaa ggtggaacca tttcatctgg agaaaagggt acatctctat
     3781 gtccttgagg gccaaatgga gctcccacgt ctgtccgggg cccaggtccc ttacctaggc
     3841 tttgggattg ggccatcatg gcctgtattg gcccttctgg ttttttctga ggcccatcta
     3901 ataccccagg attctgctgc ggtcccccac tttgtgctcc tgtgaattct ttctcatcag
     3961 gaaaaagcat gcgctggata tctctgagag tttgtaagga gcgctcccgg tgctccagct
     4021 gctcctgaga taggccatcg ggattctctc ccagtgtggg gggctcgcca ctggacactg
     4081 gtggaggcgg aggggctttg ggatctgctg aagagctatt gctcccctgg gagacagggg
     4141 tcactgccct attgttggga gttgagttgg gcccagtacc atctgggggc agtggagtgg
     4201 agctggcagg acttcctaca gaaggaatca gtttgttttc taccccagga ctctcccggt
     4261 ccaggggacg tgggggtgcg gcaggcttgg gtgctggtgc cggaatgggt ggagttggct
     4321 gcagtctggt attctgggaa gaattctggt cctggttggc cggagctggg ggctgttgtg
     4381 ggagaggttt cggatcattc cgaagggcag atatctgtgt gttcagaggc gctgtgcttc
     4441 tctctgtctt gttgttagaa atgttctgga tgtggaaaga gacgatagtt tcaacctggc
     4501 ccttcaaaac agcttctgca gctttattgg ccatctcagt agaaaacacg tacaccactt
     4561 tggctggagt cttctgagca ggtgtgggct ctgtggcagt agtttggcca tgggaggggg
     4621 tagaagacct gggggctgta gcatttgatg gggtcatcga gtgtggtgtg tgctgggaat
     4681 cctgggactt tatgtggtca gcagaattac attctttaat gtcagagtca tcgtttggag
     4741 tcccaggatc tctctgatca aaggagtcgg cggaaatact tcgctccctt ttccccttgc
     4801 ccttggcacc atttccagcc ccattcttca gccccatgct cccacctggg ccagggagtg
     4861 ctttaggggt atggccccca ctcttggagt cacaggggga tggctgggat tggctggctg
     4921 agcccccctg tttaccctga ttggagaatt tggaatccag ctgggggttt ccagatgggg
     4981 acatcactgt agggggacgg accatcacct cctgctttga cttagggcta ctctgtgtgt
     5041 ttcctgatgg agagctcctc actttagggt tactggaatg catggcgatc gctagctgca
     5101 aaaagaacaa gtagcttgta ttctatagtg tcacctaaat gggcgatctg acggttcact
     5161 aaaccagctc tgcttatata ggcctccctg gcgcgccact agttattaat agtaatcaat
     5221 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
     5281 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
     5341 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
     5401 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtccgcccc ctattgacgt
     5461 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttac gggactttcc
     5521 tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca
     5581 gtacaccaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
     5641 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
     5701 taaccccgcc ccgttgacgc aaatgggcgg taggcgtgta cggtgggagg tcaaatcgaa
     5761 ttctaatact gttgacaatt aatcatcggc atagtatatc ggcatagtat aatacgacaa
     5821 ggtgaggaac taaacccagg aggcagatca tgattgaaca agatggattg cacgcaggtt
     5881 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     5941 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     6001 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
     6061 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     6121 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     6181 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     6241 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     6301 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     6361 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     6421 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     6481 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     6541 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     6601 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgattcgaa atgaccgacc
     6661 aagcgacgcc caaccggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     6721 aggggataac gcaggaaaga acatgtcaaa catgagaatt acaacttata tcgtatgggg
     6781 ctgacttcag gtgctacatt tgaagagata aattgcactg aaatctagaa atattttatc
     6841 tgattaataa gatgatcttc ttgagatcgt tttggtctgc gcgtaatctc ttgctctgaa
     6901 aacgaaaaaa ccgccttgca gggcggtttt tcgaaggttc tctgagctac caactctttg
     6961 aaccgaggta actggcttgg aggagcgcag tcaccaaaac ttgtcctttc agtttagcct
     7021 taaccggcgc atgacttcaa gactaactcc tctaaatcaa ttaccagtgg ctgctgccag
     7081 tggtgctttt gcatgtcttt ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     7141 gcggtcggac tgaacggggg gttcgtgcat acagtccagc ttggagcgaa ctgcctaccc
     7201 ggaactgagt gtcaggcgtg gaatgagaca aacgcggcca taacagcgga atgacaccgg
     7261 taaaccgaaa ggcaggaaca ggagagcgca cgagggagcc gccaggggga aacgcctggt
     7321 atctttatag tcctgtcggg tttcgccacc actgatttga gcgtcagatt tcgtgatgct
     7381 tgtcaggggg gcggagccta tggaaaaacg gctttgccgc ggccctctca cttccctgtt
     7441 aagtatcttc ctggcatctt ccaggaaatc tccgccccgt tcgtaagcca tttccgctcg
     7501 ccgcagtcga acgaccgagc gtagcgagtc agtgagcgag gaagcggaat atatcctgta
     7561 tcacatattc tgctgacgca ccggtgcagc cttttttctc ctgccacatg aagcacttca
     7621 ctgacaccct catcagtgcc aacatagtaa gccagtatac actccgctag cgctgatgtc
     7681 cggaatcgtc acacaaaaaa ccaacacaca gatgtaatga aaataaagat attttattgc
     7741 ggccatcgtg atggctagag tctaaactcg acctacctcc ttagcgcgcg gtcgtgtact
     7801 gggccggcgg ttgccgctgt cggccagcac ctcgacgcag ccgtcggtcc atacggctgc
     7861 gctccggcga gcgatttgtg tgcgaccgac ggtgccggcc ccgctgcgga cgatggcatc
     7921 gcagcggcct tgagcccagg cagcatcgtc gaagttgccg tccacgaggc tctggtagag
     7981 ctgatccagg ccgatgcgca gcatgtaggc tcgcagacga gggctgccgg ccagctcggg
     8041 atgccggcgc tcgaagtagc gagtctgctg ctccatgcaa gccagccagg gccgccagaa
     8101 gaagatgttg gccacctcgt actgactgtc cccgaacata gcttcggacc agtcgattac
     8161 ggcggtgatg cggccgttgt cggtcaggac gttgttgctg ccgaagtcgg catggacgag
     8221 gtggcgcacc tcgggacagt cttcggccca cagcatcagt tcgtccaggg cttgagctac
     8281 gctggcggac acggtgtcgt ccatcacggt ctgccagtgg tagacatggg gatcagcaat
     8341 ggcgcaaatg aaatcccgcc aggttgtgta ctggccgatg ccttggggcc cgaatgggcc
     8401 gaagccgctg gtttgactga ggtcggcggc ggcaatagca tccatagctt cggcgacagg
     8461 ctgtaacaca gcaggcagct ctgtttcggg gaggtcttgg agagtaacgc cttgggcgcg
     8521 tctactgatg cagtatgtca ggctctcgct gaactcgccg atgtccaaca cttcggggat
     8581 gggtagtgca gcgctggcga agtggcggta cacgtagcgg tctttgtaga agccatcagc
     8641 gcagctattc acccgcagta catagccgcg tccgccgaca tcgaagctga aggctcggct
     8701 ctcttcgccc tccgacaact gcatcaggtc gctcacactg tcgaacttct cgatgagaaa
     8761 tttttcaacg ctggtagcgg tgagttcggg cttcttcatg gtggcgctag tgtcagaaga
     8821 atcgagcttt ttgcaaaagc ctaggcctcc aaaaaagcct cctcactact tctggaatag
     8881 ctcagaggcc gaggcggcct cggcctctgc ataaataaaa aaaattagtc agccatgggg
     8941 cggagaatgg gcggaactgg gcggagttag gggcgggatg ggcggagtta ggggcgggac
     9001 tatggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg
     9061 ggactttcca cacctggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc
     9121 tggggagcct ggggactttc cacaccctaa ctgacacaca ttccacagct ggttctttcc
     9181 gcctcagaag gtacctaacc aagttcctct ttcagaggtt atttcaggcc atggtgctgc
     9241 gc
//
