LOCUS       sequence-471920-        9374 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express LgBiT-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 9374)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9374)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: LgBiT-BCL9 Fusion Vector
FEATURES             Location/Qualifiers
     source          1..9374
                     /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"
     primer_bind     275..293
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     promoter        complement(5254..5272)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(5255..5272)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     enhancer        5392..5695
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     5850..5870
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        5903..5950
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5982..6776
                     /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(6036..6055)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6646..6665
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      6993..7538
                     /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     7439..7458
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7655..7672
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    7822..7870
                     /note="synthetic polyadenylation signal"
     CDS             complement(7894..8931)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             7907..7915
                     /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(8962..9319)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8976..9111
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        9128..9319
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     9276..9296
                     /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 ccccaagggg ttatgctagt tattgctcag cggtggcagc
      301 agccaactca gcttcctttc gggctttgtt agcagccgga tcagcttgca tgcctgcagg
      361 tcgactctag aggatccccg ggtaccgagc ccgaattcgt ttaaacaaac atcatgtttc
      421 ctgggttgcc aggtccttgg ctaaatccac ccatgcccac gtcagcagcc atgccccggg
      481 gcctcatctg tggggggatc atgatgttct gttggggtcc catcatgccc tgcatggaca
      541 tcatcatgcc tggagagccc acaggcccag ggtgggtgta gagcccggca ggcccccgat
      601 ccttgccagg aatcatgccc acggcagcag ctggattgct catcagggtg ggctggccgg
      661 gcattgtaga ttgtgctggt gacatcatcc gatggtgagg tcccatcatg ccttgttgga
      721 gaaaggctgg tggtctcatg gggttgtggc ctggcatgga tggagctgta ccaagaggga
      781 tgtccggagt tcccactggc cctggacctc ccatgccagg taatgctagt cccattctgg
      841 gggcttgttc gcccatcatc ccctgcatgt gtgaaaaccc aggtccagga ccctggggct
      901 gtttacggcc tggaacttcc cctcgaggga aatattgcag cgtctggctg ggcttctcag
      961 atggaataat gcgggataga tcaaactcag gtattccggt ggctccaggt cggatgacct
     1021 cctgcagatc tgggtctgta aacaccgaag gcatgctgtt ccccaggaca gtgaaggagt
     1081 caggaccccc ggggcctcca ggcttgcaaa gtgctgcatc tgctgaactt tggggcaggt
     1141 tgctggggcg gccaaggggg ccttctcctg ggaaacccat ccctcctggg aagctgccct
     1201 gccccccact ggggccattg tgagggaatg gaacctgctg tgggggagac tgtactggag
     1261 ggaagccctg ggggaagccc atccgtcctt gaggtaccat cggtggctcc tgggacccat
     1321 gccccatgat aggattgtgt gacatcaagc caggccccat tgggacccca tgaggaggta
     1381 tgttgggtcc cacggcattt ggagagggca tctgattgga gtgagaaagt ggctgggtca
     1441 ttcccattgg gctgagggtt ggcatcggaa ccacggggtt tggacctgaa attcgaggat
     1501 tctgtgtatt aatgcccatt cctggcatat tattcattga tggcaagttg ggagaacgag
     1561 ctggagggga gtcgtcatct gagctggcca cagtcttgat agcatcatgg tataacgggg
     1621 tggaactggg cattgcaaac ttggacattc gagacatcat aatagagagt gggttctggg
     1681 aaagggttgg ctctggaggc atggtataag gtgtactaga gggaagactt ccagggatat
     1741 tcacagaggc aggctggctg gctgtaggag gtggggggcc acctgactct acatttccca
     1801 gcatggctgg ggaggccatg gtgaggggtg ctttatggtt tggagggatc ccaggactct
     1861 gaaggggagg ttttggagaa gaggtccatc caggtgacgg ggcaggaagt gatggagact
     1921 tgaggtggac aggtgaagca gcagcagacc ccaaaacagg gggggactta atggaagcag
     1981 cagcagctgg gcccgccagc atgcctgcca gctgcgatgg agtctggggg gacttgaggt
     2041 tccccgaggg cgagcccagc attggtgact ggacttggtg catcgtggga gacttcagag
     2101 ggttaatgcc tggggaatgc acctggctgc ctgccacaga tatatccaag ggcttccgcc
     2161 ccaggccgcg ctgaactgga ggagctgtgt tgaggctggt ggggttactg gaagggttga
     2221 gaggtagtgg tggcatatga ctgagccggc tgttagtcct ctggtcgggc ccaattggtt
     2281 ctctgagatt ccgcaagcca ctgttgctgc ctggaccctg agacatggga aggaatggtc
     2341 tggggcccat gccatactcc tgctgggggt gctcaccaaa tggcagtggc accatcttct
     2401 gctgagcagg cagcatgtct gagccacctg ggcgtaattt cagcatctcc tcagggcccg
     2461 ccccagcctc tctcatcttc tgaggtatca tctgagagtt ggatcccatg ttgacattta
     2521 gattgacatc tcccttcatc ccactaggaa ccatcccaaa ctcaagttcc cgaccagggc
     2581 ccatctgtgg gggaattcct tttggaaagt cacccctaga aggactcaga gggccctcaa
     2641 ctggtattcg agggaaaatg gggttattcc caggctccat gtggcgctgg gagcctggaa
     2701 tcatcctgtt catctccatg ctgggcctga tgccttccat ggccatccct ggggggagac
     2761 ccagctgttt ctctgccagc tgctgctgaa acatctcttc agacaatcct tgggggtttg
     2821 ggaagcgttc ccctcggcca ggaccgctga aaatgccctg gccaggagga aaatttcgac
     2881 catctgggat ttttggcaca tcatctggcc aactgactcc agaaagacct ggtctagatg
     2941 cagggttggg gacattcggc ccttccattt cagagtttat catgcctgca aatccaggga
     3001 ggcgcatctg gctccctggc atgttggggt ggggagccat gcccctcggg ggcagagaat
     3061 gtggcatgtt gataccatca gaaaatggct ctgtaccccc aggtgcccag ccttcactag
     3121 gggtcatctg gtatggaggg gggggtcctc ggaccactcc ccgaggcccg tgctgatgga
     3181 ccatcatgtc ctggagggaa cactgctgga caaccacttg ttcctgcttc ctcctcttct
     3241 cttcataaaa ctcctgctgc agtttcagcc acgctatctg ctcgggagtc atatggtcaa
     3301 ggtggtcggg tcctatggtc ccagactggg agttcataga aggtggaacc atttcatctg
     3361 gagaaaaggg tacatctcta tgtccttgag ggccaaatgg agctcccacg tctgtccggg
     3421 gcccaggtcc cttacctagg ctttgggatt gggccatcat ggcctgtatt ggcccttctg
     3481 gttttttctg aggcccatct aataccccag gattctgctg cggtccccca ctttgtgctc
     3541 ctgtgaattc tttctcatca ggaaaaagca tgcgctggat atctctgaga gtttgtaagg
     3601 agcgctcccg gtgctccagc tgctcctgag ataggccatc gggattctct cccagtgtgg
     3661 ggggctcgcc actggacact ggtggaggcg gaggggcttt gggatctgct gaagagctat
     3721 tgctcccctg ggagacaggg gtcactgccc tattgttggg agttgagttg ggcccagtac
     3781 catctggggg cagtggagtg gagctggcag gacttcctac agaaggaatc agtttgtttt
     3841 ctaccccagg actctcccgg tccaggggac gtgggggtgc ggcaggcttg ggtgctggtg
     3901 ccggaatggg tggagttggc tgcagtctgg tattctggga agaattctgg tcctggttgg
     3961 ccggagctgg gggctgttgt gggagaggtt tcggatcatt ccgaagggca gatatctgtg
     4021 tgttcagagg cgctgtgctt ctctctgtct tgttgttaga aatgttctgg atgtggaaag
     4081 agacgatagt ttcaacctgg cccttcaaaa cagcttctgc agctttattg gccatctcag
     4141 tagaaaacac gtacaccact ttggctggag tcttctgagc aggtgtgggc tctgtggcag
     4201 tagtttggcc atgggagggg gtagaagacc tgggggctgt agcatttgat ggggtcatcg
     4261 agtgtggtgt gtgctgggaa tcctgggact ttatgtggtc agcagaatta cattctttaa
     4321 tgtcagagtc atcgtttgga gtcccaggat ctctctgatc aaaggagtcg gcggaaatac
     4381 ttcgctccct tttccccttg cccttggcac catttccagc cccattcttc agccccatgc
     4441 tcccacctgg gccagggagt gctttagggg tatggccccc actcttggag tcacaggggg
     4501 atggctggga ttggctggct gagcccccct gtttaccctg attggagaat ttggaatcca
     4561 gctgggggtt tccagatggg gacatcactg tagggggacg gaccatcacc tcctgctttg
     4621 acttagggct actctgtgtg tttcctgatg gagagctcct cactttaggg ttactggaat
     4681 gcatggcgat cgcaccgctc gagcctccac ctccgctccc gccaccaccg gagctcccac
     4741 tgttgatggt tactcggaac agcatggagc cgtcgggggt gatcaggcgc tcgtcgataa
     4801 ttttgttgcc gttccacagg gtccctgtta cagtgatctt tttgccgtcg aacacggcga
     4861 tgccttcata cggccgtccg aaatagttca gcatgttcgg cgtaaccccg tcgattacca
     4921 gtgtgccata gggcaggatc accttaaagt gatgatcatc cacagggtac accaccttaa
     4981 acacctcttc gatctgggcc atttggtcgg cgctcagacc ttcatacggg atgatgacat
     5041 ggatgtcgat cttcagggca ttttcaccgc tccggacaat cctttggatc ggagttacgg
     5101 acacggcgag attctgcagc aaactggaca cacctccctg ttcaaggact tggtccaggt
     5161 tgtaggcggc tgtctgttcc cagtccccaa cgaaatcttc gagtgtgaag accatggtga
     5221 gcgctagctg caaaaagaac aagtagcttg tattctatag tgtcacctaa atgggcgatc
     5281 tgacggttca ctaaaccagc tctgcttata taggcctccc tggcgcgcca ctagttatta
     5341 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
     5401 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
     5461 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
     5521 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtccgcc
     5581 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
     5641 acgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
     5701 gcggttttgg cagtacacca atgggcgtgg atagcggttt gactcacggg gatttccaag
     5761 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
     5821 aaaatgtcgt aataaccccg ccccgttgac gcaaatgggc ggtaggcgtg tacggtggga
     5881 ggtcaaatcg aattctaata ctgttgacaa ttaatcatcg gcatagtata tcggcatagt
     5941 ataatacgac aaggtgagga actaaaccca ggaggcagat catgattgaa caagatggat
     6001 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     6061 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     6121 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc
     6181 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     6241 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     6301 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     6361 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     6421 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     6481 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga
     6541 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     6601 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     6661 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     6721 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgattcg
     6781 aaatgaccga ccaagcgacg cccaaccggt atcagctcac tcaaaggcgg taatacggtt
     6841 atccacagaa tcaggggata acgcaggaaa gaacatgtca aacatgagaa ttacaactta
     6901 tatcgtatgg ggctgacttc aggtgctaca tttgaagaga taaattgcac tgaaatctag
     6961 aaatatttta tctgattaat aagatgatct tcttgagatc gttttggtct gcgcgtaatc
     7021 tcttgctctg aaaacgaaaa aaccgccttg cagggcggtt tttcgaaggt tctctgagct
     7081 accaactctt tgaaccgagg taactggctt ggaggagcgc agtcaccaaa acttgtcctt
     7141 tcagtttagc cttaaccggc gcatgacttc aagactaact cctctaaatc aattaccagt
     7201 ggctgctgcc agtggtgctt ttgcatgtct ttccgggttg gactcaagac gatagttacc
     7261 ggataaggcg cagcggtcgg actgaacggg gggttcgtgc atacagtcca gcttggagcg
     7321 aactgcctac ccggaactga gtgtcaggcg tggaatgaga caaacgcggc cataacagcg
     7381 gaatgacacc ggtaaaccga aaggcaggaa caggagagcg cacgagggag ccgccagggg
     7441 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca ccactgattt gagcgtcaga
     7501 tttcgtgatg cttgtcaggg gggcggagcc tatggaaaaa cggctttgcc gcggccctct
     7561 cacttccctg ttaagtatct tcctggcatc ttccaggaaa tctccgcccc gttcgtaagc
     7621 catttccgct cgccgcagtc gaacgaccga gcgtagcgag tcagtgagcg aggaagcgga
     7681 atatatcctg tatcacatat tctgctgacg caccggtgca gccttttttc tcctgccaca
     7741 tgaagcactt cactgacacc ctcatcagtg ccaacatagt aagccagtat acactccgct
     7801 agcgctgatg tccggaatcg tcacacaaaa aaccaacaca cagatgtaat gaaaataaag
     7861 atattttatt gcggccatcg tgatggctag agtctaaact cgacctacct ccttagcgcg
     7921 cggtcgtgta ctgggccggc ggttgccgct gtcggccagc acctcgacgc agccgtcggt
     7981 ccatacggct gcgctccggc gagcgatttg tgtgcgaccg acggtgccgg ccccgctgcg
     8041 gacgatggca tcgcagcggc cttgagccca ggcagcatcg tcgaagttgc cgtccacgag
     8101 gctctggtag agctgatcca ggccgatgcg cagcatgtag gctcgcagac gagggctgcc
     8161 ggccagctcg ggatgccggc gctcgaagta gcgagtctgc tgctccatgc aagccagcca
     8221 gggccgccag aagaagatgt tggccacctc gtactgactg tccccgaaca tagcttcgga
     8281 ccagtcgatt acggcggtga tgcggccgtt gtcggtcagg acgttgttgc tgccgaagtc
     8341 ggcatggacg aggtggcgca cctcgggaca gtcttcggcc cacagcatca gttcgtccag
     8401 ggcttgagct acgctggcgg acacggtgtc gtccatcacg gtctgccagt ggtagacatg
     8461 gggatcagca atggcgcaaa tgaaatcccg ccaggttgtg tactggccga tgccttgggg
     8521 cccgaatggg ccgaagccgc tggtttgact gaggtcggcg gcggcaatag catccatagc
     8581 ttcggcgaca ggctgtaaca cagcaggcag ctctgtttcg gggaggtctt ggagagtaac
     8641 gccttgggcg cgtctactga tgcagtatgt caggctctcg ctgaactcgc cgatgtccaa
     8701 cacttcgggg atgggtagtg cagcgctggc gaagtggcgg tacacgtagc ggtctttgta
     8761 gaagccatca gcgcagctat tcacccgcag tacatagccg cgtccgccga catcgaagct
     8821 gaaggctcgg ctctcttcgc cctccgacaa ctgcatcagg tcgctcacac tgtcgaactt
     8881 ctcgatgaga aatttttcaa cgctggtagc ggtgagttcg ggcttcttca tggtggcgct
     8941 agtgtcagaa gaatcgagct ttttgcaaaa gcctaggcct ccaaaaaagc ctcctcacta
     9001 cttctggaat agctcagagg ccgaggcggc ctcggcctct gcataaataa aaaaaattag
     9061 tcagccatgg ggcggagaat gggcggaact gggcggagtt aggggcggga tgggcggagt
     9121 taggggcggg actatggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc
     9181 tggggagcct ggggactttc cacacctggt tgctgactaa ttgagatgca tgctttgcat
     9241 acttctgcct gctggggagc ctggggactt tccacaccct aactgacaca cattccacag
     9301 ctggttcttt ccgcctcaga aggtacctaa ccaagttcct ctttcagagg ttatttcagg
     9361 ccatggtgct gcgc
//
