LOCUS       sequence-474687-        7214 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express LgBiT-BCL6 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 7214)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7214)
  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            "Unpublished"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-BCL6 Vector
FEATURES             Location/Qualifiers
     source          1..7214
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        50..68
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     50..67
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2869..2887)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3064..3145
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3101..3120)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3217..3574
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3217..3408
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3240..3260)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3425..3560
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3605..4642
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             4621..4629
                     /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)"
     polyA_signal    4666..4714
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(4864..4881)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4998..5543)
                     /direction=LEFT
                     /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     complement(5078..5097)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5760..6554)
                     /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(5871..5890)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     6481..6500
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(6586..6633)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     primer_bind     complement(6666..6686)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        6841..7144
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 agggaggcct atataagcag agctggttta gtgaaccgtc agatcgccca tttaggtgac
       61 actatagaat acaagctact tgttcttttt gcagctagcg ctcaccatgg tcttcacact
      121 cgaagatttc gttggggact gggaacagac agccgcctac aacctggacc aagtccttga
      181 acagggaggt gtgtccagtt tgctgcagaa tctcgccgtg tccgtaactc cgatccaaag
      241 gattgtccgg agcggtgaaa atgccctgaa gatcgacatc catgtcatca tcccgtatga
      301 aggtctgagc gccgaccaaa tggcccagat cgaagaggtg tttaaggtgg tgtaccctgt
      361 ggatgatcat cactttaagg tgatcctgcc ctatggcaca ctggtaatcg acggggttac
      421 gccgaacatg ctgaactatt tcggacggcc gtatgaaggc atcgccgtgt tcgacggcaa
      481 aaagatcact gtaacaggga ccctgtggaa cggcaacaaa attatcgacg agcgcctgat
      541 cacccccgac ggctccatgc tgttccgagt aaccatcaac agtgggagct ccggtggtgg
      601 cgggagcgga ggtggaggct cgagcggtgc gatcgccatg gcctcgccgg ctgacagctg
      661 tatccagttc acccgccatg ccagtgatgt tcttctcaac cttaatcgtc tccggagtcg
      721 agacatcttg actgatgttg tcattgttgt gagccgtgag cagtttagag cccataaaac
      781 ggtcctcatg gcctgcagtg gcctgttcta tagcatcttt acagaccagt tgaaatgcaa
      841 ccttagtgtg atcaatctag atcctgagat caaccctgag ggattctgca tcctcctgga
      901 cttcatgtac acatctcggc tcaatttgcg ggagggcaac atcatggctg tgatggccac
      961 ggctatgtac ctgcagatgg agcatgttgt ggacacttgc cggaagttta ttaaggccag
     1021 tgaagcagag atggtttctg ccatcaagcc tcctcgtgaa gagttcctca acagccggat
     1081 gctgatgccc caagacatca tggcctatcg gggtcgtgag gtggtggaga acaacctgcc
     1141 actgaggagc gcccctgggt gtgagagcag agcctttgcc cccagcctgt acagtggcct
     1201 gtccacaccg ccagcctctt attccatgta cagccacctc cctgtcagca gcctcctctt
     1261 ctccgatgag gagtttcggg atgtccggat gcctgtggcc aaccccttcc ccaaggagcg
     1321 ggcactccca tgtgatagtg ccaggccagt ccctggtgag tacagccggc cgactttgga
     1381 ggtgtccccc aatgtgtgcc acagcaatat ctattcaccc aaggaaacaa tcccagaaga
     1441 ggcacgaagt gatatgcact acagtgtggc tgagggcctc aaacctgctg ccccctcagc
     1501 ccgaaatgcc ccctacttcc cttgtgacaa ggccagcaaa gaagaagaga gaccctcctc
     1561 ggaagatgag attgccctgc atttcgagcc ccccaatgca cccctgaacc ggaagggtct
     1621 ggttagtcca cagagccccc agaaatctga ctgccagccc aactcgccca cagagtcctg
     1681 cagcagtaag aatgcctgca tcctccaggc ttctggctcc cctccagcca agagccccac
     1741 tgaccccaaa gcctgcaact ggaagaaata caagttcatc gtgctcaaca gcctcaacca
     1801 gaatgccaaa ccagaggggc ctgagcaggc tgagctgggc cgcctttccc cacgagccta
     1861 cacggcccca cctgcctgcc agccacccat ggagcctgag aaccttgacc tccagtcccc
     1921 aaccaagctg agtgccagcg gggaggactc caccatccca caagccagcc ggctcaataa
     1981 catcgttaac aggtccatga cgggctctcc ccgcagcagc agcgagagcc actcaccact
     2041 ctacatgcac cccccgaagt gcacgtcctg cggctctcag tccccacagc atgcagagat
     2101 gtgcctccac accgctggcc ccacgttccc tgaggagatg ggagagaccc agtctgagta
     2161 ctcagattct agctgtgaga acggggcctt cttctgcaat gagtgtgact gccgcttctc
     2221 tgaggaggcc tcactcaaga ggcacacgct gcagacccac agtgacaaac cctacaagtg
     2281 tgaccgctgc caggcctcct tccgctacaa gggcaacctc gccagccaca agaccgtcca
     2341 taccggtgag aaaccctatc gttgcaacat ctgtggggcc cagttcaacc ggccagccaa
     2401 cctgaaaacc cacactcgaa ttcactctgg agagaagccc tacaaatgcg aaacctgcgg
     2461 agccagattt gtacaggtgg cccacctccg tgcccatgtg cttatccaca ctggtgagaa
     2521 gccctatccc tgtgaaatct gtggcacccg tttccggcac cttcagactc tgaagagcca
     2581 cctgcgaatc cacacaggag agaaacctta ccattgtgag aagtgtaacc tgcatttccg
     2641 tcacaaaagc cagctgcgac ttcacttgcg ccagaagcat ggcgccatca ccaacaccaa
     2701 ggtgcaatac cgcgtgtcag ccactgacct gcctccggag ctccccaaag cctgcgttta
     2761 aacgaattcg ggctcggtac ccggggatcc tctagagtcg acctgcaggc atgcaagctg
     2821 atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat
     2881 aactagcata accccttggg gaaacaccta ggctctcgta cgaagtcagc tgataacgtc
     2941 gacgtattgc ggccgcggat cataatcagc cataccacat ttgtagaggt tttacttgct
     3001 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt
     3061 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     3121 acaaataaag catttttttc actgccgacg tcacgcgtct tgcgcagcac catggcctga
     3181 aataacctct gaaagaggaa cttggttagg taccttctga ggcggaaaga accagctgtg
     3241 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3301 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     3361 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     3421 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     3481 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     3541 aggaggcttt tttggaggcc taggcttttg caaaaagctc gattcttctg acactagcgc
     3601 caccatgaag aagcccgaac tcaccgctac cagcgttgaa aaatttctca tcgagaagtt
     3661 cgacagtgtg agcgacctga tgcagttgtc ggagggcgaa gagagccgag ccttcagctt
     3721 cgatgtcggc ggacgcggct atgtactgcg ggtgaatagc tgcgctgatg gcttctacaa
     3781 agaccgctac gtgtaccgcc acttcgccag cgctgcacta cccatccccg aagtgttgga
     3841 catcggcgag ttcagcgaga gcctgacata ctgcatcagt agacgcgccc aaggcgttac
     3901 tctccaagac ctccccgaaa cagagctgcc tgctgtgtta cagcctgtcg ccgaagctat
     3961 ggatgctatt gccgccgccg acctcagtca aaccagcggc ttcggcccat tcgggcccca
     4021 aggcatcggc cagtacacaa cctggcggga tttcatttgc gccattgctg atccccatgt
     4081 ctaccactgg cagaccgtga tggacgacac cgtgtccgcc agcgtagctc aagccctgga
     4141 cgaactgatg ctgtgggccg aagactgtcc cgaggtgcgc cacctcgtcc atgccgactt
     4201 cggcagcaac aacgtcctga ccgacaacgg ccgcatcacc gccgtaatcg actggtccga
     4261 agctatgttc ggggacagtc agtacgaggt ggccaacatc ttcttctggc ggccctggct
     4321 ggcttgcatg gagcagcaga ctcgctactt cgagcgccgg catcccgagc tggccggcag
     4381 ccctcgtctg cgagcctaca tgctgcgcat cggcctggat cagctctacc agagcctcgt
     4441 ggacggcaac ttcgacgatg ctgcctgggc tcaaggccgc tgcgatgcca tcgtccgcag
     4501 cggggccggc accgtcggtc gcacacaaat cgctcgccgg agcgcagccg tatggaccga
     4561 cggctgcgtc gaggtgctgg ccgacagcgg caaccgccgg cccagtacac gaccgcgcgc
     4621 taaggaggta ggtcgagttt agactctagc catcacgatg gccgcaataa aatatcttta
     4681 ttttcattac atctgtgtgt tggttttttg tgtgacgatt ccggacatca gcgctagcgg
     4741 agtgtatact ggcttactat gttggcactg atgagggtgt cagtgaagtg cttcatgtgg
     4801 caggagaaaa aaggctgcac cggtgcgtca gcagaatatg tgatacagga tatattccgc
     4861 ttcctcgctc actgactcgc tacgctcggt cgttcgactg cggcgagcgg aaatggctta
     4921 cgaacggggc ggagatttcc tggaagatgc caggaagata cttaacaggg aagtgagagg
     4981 gccgcggcaa agccgttttt ccataggctc cgcccccctg acaagcatca cgaaatctga
     5041 cgctcaaatc agtggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     5101 ggcggctccc tcgtgcgctc tcctgttcct gcctttcggt ttaccggtgt cattccgctg
     5161 ttatggccgc gtttgtctca ttccacgcct gacactcagt tccgggtagg cagttcgctc
     5221 caagctggac tgtatgcacg aaccccccgt tcagtccgac cgctgcgcct tatccggtaa
     5281 ctatcgtctt gagtccaacc cggaaagaca tgcaaaagca ccactggcag cagccactgg
     5341 taattgattt agaggagtta gtcttgaagt catgcgccgg ttaaggctaa actgaaagga
     5401 caagttttgg tgactgcgct cctccaagcc agttacctcg gttcaaagag ttggtagctc
     5461 agagaacctt cgaaaaaccg ccctgcaagg cggttttttc gttttcagag caagagatta
     5521 cgcgcagacc aaaacgatct caagaagatc atcttattaa tcagataaaa tatttctaga
     5581 tttcagtgca atttatctct tcaaatgtag cacctgaagt cagccccata cgatataagt
     5641 tgtaattctc atgtttgaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     5701 tattaccgcc tttgagtgag ctgataccgg ttgggcgtcg cttggtcggt catttcgaat
     5761 cagaagaact cgtcaagaag gcgatagaag gcgatgcgct gcgaatcggg agcggcgata
     5821 ccgtaaagca cgaggaagcg gtcagcccat tcgccgccaa gctcttcagc aatatcacgg
     5881 gtagccaacg ctatgtcctg atagcggtcc gccacaccca gccggccaca gtcgatgaat
     5941 ccagaaaagc ggccattttc caccatgata ttcggcaagc aggcatcgcc atgggtcacg
     6001 acgagatcct cgccgtcggg catgcgcgcc ttgagcctgg cgaacagttc ggctggcgcg
     6061 agcccctgat gctcttcgtc cagatcatcc tgatcgacaa gaccggcttc catccgagta
     6121 cgtgctcgct cgatgcgatg tttcgcttgg tggtcgaatg ggcaggtagc cggatcaagc
     6181 gtatgcagcc gccgcattgc atcagccatg atggatactt tctcggcagg agcaaggtga
     6241 gatgacagga gatcctgccc cggcacttcg cccaatagca gccagtccct tcccgcttca
     6301 gtgacaacgt cgagcacagc tgcgcaagga acgcccgtcg tggccagcca cgatagccgc
     6361 gctgcctcgt cctgcagttc attcagggca ccggacaggt cggtcttgac aaaaagaacc
     6421 gggcgcccct gcgctgacag ccggaacacg gcggcatcag agcagccgat tgtctgttgt
     6481 gcccagtcat agccgaatag cctctccacc caagcggccg gagaacctgc gtgcaatcca
     6541 tcttgttcaa tcatgatctg cctcctgggt ttagttcctc accttgtcgt attatactat
     6601 gccgatatac tatgccgatg attaattgtc aacagtatta gaattcgatt tgacctccca
     6661 ccgtacacgc ctaccgccca tttgcgtcaa cggggcgggg ttattacgac attttggaaa
     6721 gtcccgttga ttttggtgcc aaaacaaact cccattgacg tcaatggggt ggagacttgg
     6781 aaatccccgt gagtcaaacc gctatccacg cccattggtg tactgccaaa accgcatcac
     6841 catggtaata gcgatgacta atacgtagat gtactgccaa gtaggaaagt cccgtaaggt
     6901 catgtactgg gcataatgcc aggcgggcca tttaccgtca ttgacgtcaa tagggggcgg
     6961 acttggcata tgatacactt gatgtactgc caagtgggca gtttaccgta aatactccac
     7021 ccattgacgt caatggaaag tccctattgg cgttactatg ggaacatacg tcattattga
     7081 cgtcaatggg cgggggtcgt tgggcggtca gccaggcggg ccatttaccg taagttatgt
     7141 aacgcggaac tccatatatg ggctatgaac taatgacccc gtaattgatt actattaata
     7201 actagtggcg cgcc
//
