LOCUS       sequence-475217-        8852 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express LgBiT-BRD4 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 8852)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8852)
  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            "Unpublished"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-BRD4 Fusion Vector
FEATURES             Location/Qualifiers
     source          1..8852
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..721
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     672..692
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          857..989
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     972..991
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1033..1052
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1033..1051
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3807..3824
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5795..5813)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    5850..5971
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5861..5880)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5915..5934
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        6216..6573
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6216..6407
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6239..6259)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6424..6559
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        6588..6635
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             6667..7461
                     /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(6721..6740)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7331..7350
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7525..7573
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(7812..8400)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7892..7911)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    8516..8799
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      661 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      721 tggtttagtg aaccgtcaga tcactagaag ctttattgcg gtagtttatc acagttaaat
      781 tgctaacgca gtcagtgctt ctgacacaac agtctcgaac ttaagctgca gaagttggtc
      841 gtgaggcact gggcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa
      901 actgggcttg tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac
      961 tgacatccac tttgcctttc tctccacagg tgtccactcc cagttcaatt acagctctta
     1021 aggctagagt attaatacga ctcactatag ggctagcgct caccatggtc ttcacactcg
     1081 aagatttcgt tggggactgg gaacagacag ccgcctacaa cctggaccaa gtccttgaac
     1141 agggaggtgt gtccagtttg ctgcagaatc tcgccgtgtc cgtaactccg atccaaagga
     1201 ttgtccggag cggtgaaaat gccctgaaga tcgacatcca tgtcatcatc ccgtatgaag
     1261 gtctgagcgc cgaccaaatg gcccagatcg aagaggtgtt taaggtggtg taccctgtgg
     1321 atgatcatca ctttaaggtg atcctgccct atggcacact ggtaatcgac ggggttacgc
     1381 cgaacatgct gaactatttc ggacggccgt atgaaggcat cgccgtgttc gacggcaaaa
     1441 agatcactgt aacagggacc ctgtggaacg gcaacaaaat tatcgacgag cgcctgatca
     1501 cccccgacgg ctccatgctg ttccgagtaa ccatcaacag tgggagctcc ggtggtggcg
     1561 ggagcggagg tggaggctcg agcggtgcga tcgccatgtc tgcggagagc ggccctggga
     1621 cgagattgag aaatctgcca gtaatggggg atggactaga aacttcccaa atgtctacaa
     1681 cacaggccca ggcccaaccc cagccagcca acgcagccag caccaacccc ccgcccccag
     1741 agacctccaa ccctaacaag cccaagaggc agaccaacca actgcaatac ctgctcagag
     1801 tggtgctcaa gacactatgg aaacaccagt ttgcatggcc tttccagcag cctgtggatg
     1861 ccgtcaagct gaacctccct gattactata agatcattaa aacgcctatg gatatgggaa
     1921 caataaagaa gcgcttggaa aacaactatt actggaatgc tcaggaatgt atccaggact
     1981 tcaacactat gtttacaaat tgttacatct acaacaagcc tggagatgac atagtcttaa
     2041 tggcagaagc tctggaaaag ctcttcttgc aaaaaataaa tgagctaccc acagaagaaa
     2101 ccgagatcat gatagtccag gcaaaaggaa gaggacgtgg gaggaaagaa acagggacag
     2161 caaaacctgg cgtttccacg gtaccaaaca caactcaagc atcgactcct ccgcagaccc
     2221 agacccctca gccgaatcct cctcctgtgc aggccacgcc tcaccccttc cctgccgtca
     2281 ccccggacct catcgtccag acccctgtca tgacagtggt gcctccccag ccactgcaga
     2341 cgcccccgcc agtgcccccc cagccacaac ccccacccgc tccagctccc cagcccgtac
     2401 agagccaccc acccatcatc gcggccaccc cacagcctgt gaagacaaag aagggagtga
     2461 agaggaaagc agacaccacc acccccacca ccattgaccc cattcacgag ccaccctcgc
     2521 tgcccccgga gcccaagacc accaagctgg gccagcggcg ggagagcagc cggcctgtga
     2581 aacctccaaa gaaggacgtg cccgactctc agcagcaccc agcaccagag aagagcagca
     2641 aggtctcgga gcagctcaag tgctgcagcg gcatcctcaa ggagatgttt gccaagaagc
     2701 acgccgccta cgcctggccc ttctacaagc ctgtggacgt ggaggcactg ggcctacacg
     2761 actactgtga catcatcaag caccccatgg acatgagcac aatcaagtct aaactggagg
     2821 cccgtgagta ccgtgatgct caggagtttg gtgctgacgt ccgattgatg ttctccaact
     2881 gctataagta caaccctcct gaccatgagg tggtggccat ggcccgcaag ctccaggatg
     2941 tgttcgaaat gcgctttgcc aagatgccgg acgagcctga ggagccagtg gtggccgtgt
     3001 cctccccggc agtgccccct cccaccaagg ttgtggcccc gccctcatcc agcgacagca
     3061 gcagcgatag ctcctcggac agtgacagtt cgactgatga ctctgaggag gagcgagccc
     3121 agcggctggc tgagctccag gagcagctca aagccgtgca cgagcagctt gcagccctct
     3181 ctcagcccca gcagaacaaa ccaaagaaaa aggagaaaga caagaaggaa aagaaaaaag
     3241 aaaagcacaa aaggaaagag gaagtggaag agaataaaaa aagcaaagcc aaggaacctc
     3301 ctcctaaaaa gacgaagaaa aataatagca gcaacagcaa tgtgagcaag aaggagccag
     3361 cgcccatgaa gagcaagccc cctcccacgt atgagtcgga ggaagaggac aagtgcaagc
     3421 ctatgtccta tgaggagaag cggcagctca gcttggacat caacaagctc cccggcgaga
     3481 agctgggccg cgtggtgcac atcatccagt cacgggagcc ctccctgaag aattccaacc
     3541 ccgacgagat tgaaatcgac tttgagaccc tgaagccgtc cacactgcgt gagctggagc
     3601 gctatgtcac ctcctgtttg cggaagaaaa ggaaacctca agctgagaaa gttgatgtga
     3661 ttgccggctc ctccaagatg aagggcttct cgtcctcaga gtcggagagc tccagtgagt
     3721 ccagctcctc tgacagcgaa gactccgaaa cagagatggc tccgaagtca aaaaagaagg
     3781 ggcaccccgg gagggagcag aagaagcacc atcatcacca ccatcagcag atgcagcagg
     3841 ccccggctcc tgtgccccag cagccgcccc cgcctcccca gcagccccca ccgcctccac
     3901 ctccgcagca gcaacagcag ccgccacccc cgcctccccc accctccatg ccgcagcagg
     3961 cagccccggc gatgaagtcc tcgcccccac ccttcattgc cacccaggtg cccgtcctgg
     4021 agccccagct cccaggcagc gtctttgacc ccatcggcca cttcacccag cccatcctgc
     4081 acctgccgca gcctgagctg ccccctcacc tgccccagcc gcctgagcac agcactccac
     4141 cccatctcaa ccagcacgca gtggtctctc ctccagcttt gcacaacgca ctaccccagc
     4201 agccatcacg gcccagcaac cgagccgctg ccctgcctcc caagcccgcc cggcccccag
     4261 ccgtgtcacc agccttgacc caaacacccc tgctcccaca gccccccatg gcccaacccc
     4321 cccaagtgct gctggaggat gaagagccac ctgccccacc cctcacctcc atgcagatgc
     4381 agctgtacct gcagcagctg cagaaggtgc agccccctac gccgctactc ccttccgtga
     4441 aggtgcagtc ccagccccca ccccccctgc cgcccccacc ccacccctct gtgcagcagc
     4501 agctgcagca gcagccgcca ccacccccac caccccagcc ccagcctcca ccccagcagc
     4561 agcatcagcc ccctccacgg cccgtgcact tgcagcccat gcagttttcc acccacatcc
     4621 aacagccccc gccaccccag ggccagcagc ccccccatcc gcccccaggc cagcagccac
     4681 ccccgccgca gcctgccaag cctcagcaag tcatccagca ccaccattca ccccggcacc
     4741 acaagtcgga cccctactca accggtcacc tccgcgaagc cccctccccg cttatgatac
     4801 attcccccca gatgtcacag ttccagagcc tgacccacca gtctccaccc cagcaaaacg
     4861 tccagcctaa gaaacaggag ctgcgtgctg cctccgtggt ccagccccag cccctcgtgg
     4921 tggtgaagga ggagaagatc cactcaccca tcatccgcag cgagcccttc agcccctcgc
     4981 tgcggccgga gccccccaag cacccggaga gcatcaaggc ccccgtccac ctgccccagc
     5041 ggccggaaat gaagcctgtg gatgtcggga ggcctgtgat ccggccccca gagcagaacg
     5101 caccgccacc aggggcccct gacaaggaca aacagaaaca ggagccgaag actccagttg
     5161 cgcccaaaaa ggacctgaaa atcaagaaca tgggctcctg ggccagccta gtgcagaagc
     5221 atccgaccac cccctcctcc acagccaagt catccagcga cagcttcgag cagttccgcc
     5281 gcgccgctcg ggagaaagag gagcgtgaga aggccctgaa ggctcaggcc gagcacgctg
     5341 agaaggagaa ggagcggctg cggcaggagc gcatgaggag ccgagaggac gaggatgcgc
     5401 tggagcaggc ccggcgggcc catgaggagg cacgtcggcg ccaggagcag cagcagcagc
     5461 agcgccagga gcaacagcag cagcagcaac agcaagcagc tgcggtggct gccgccgcca
     5521 ccccacaggc ccagagctcc cagccccagt ccatgctgga ccagcagagg gagttggccc
     5581 ggaagcggga gcaggagcga agacgccggg aagccatggc agctaccatt gacatgaatt
     5641 tccagagtga tctattgtca atatttgaag aaaatctttt cgtttaaacg aattcgggct
     5701 cggtacccgg ggatcctcta gagtcgacct gcaggcatgc aagctgatcc ggctgctaac
     5761 aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc
     5821 cttggggcgg ccgcttcgag cagacatgat aagatacatt gatgagtttg gacaaaccac
     5881 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     5941 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
     6001 tcaggttcag ggggagatgt gggaggtttt tttaagcaag taaaacctct acaaatgtgg
     6061 taaaatcgaa ttttaacaaa atattaacgc ttacaatttc ctgatgcggt attttctcct
     6121 tacgcatctg tgcggtattt cacaccgcat acgcggatct gcgcagcacc atggcctgaa
     6181 ataacctctg aaagaggaac ttggttaggt accttctgag gcggaaagaa ccagctgtgg
     6241 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     6301 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     6361 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     6421 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     6481 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     6541 ggaggctttt ttggaggcct aggcttttgc aaaaagctta attaactgtt gacaattaat
     6601 catcggcata gtatatcggc atagtataat acgacaaggt gaggaactaa acccaggagg
     6661 cagatcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     6721 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     6781 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     6841 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     6901 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     6961 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     7021 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     7081 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     7141 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     7201 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     7261 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     7321 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     7381 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     7441 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     7501 ccaacctgcc atcacgatgg ccgcaataaa atatctttat tttcattaca tctgtgtgtt
     7561 ggttttttgt gtgaatcgat agcgataagg atcctctttg cgcttgcgtt ttcccttgtc
     7621 cagatagccc agtagctgac attcatccgg ggtcagcacc gtttctgcgg actggctttc
     7681 tacccggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     7741 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     7801 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     7861 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     7921 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     7981 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     8041 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     8101 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     8161 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     8221 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     8281 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     8341 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatttcaa
     8401 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     8461 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt atagtccgga
     8521 aatacaggaa cgcacgctgg atggcccttc gctgggatgg tgaaaccatg aaaaatggca
     8581 gcttcagtgg attaagtggg ggtaatgtgg cctgtaccct ctggttgcat aggtattcat
     8641 acggttaaaa tttatcaggc gcgattgcgg cagtttttcg ggtggtttgt tgccattttt
     8701 acctgtctgc tgccgtgatc gcgctgaacg cgttttagcg gtgcgtacaa ttaagggatt
     8761 atggtaaatc cacttactgt ctgccctcgt agccatcgag ataaaccgca gtactccggc
     8821 cacgatgcgt ccggcgtaga ggatcgagat ct
//
