LOCUS       sequence-471907-        5594 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express LgBiT-CHMP4B 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 5594)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5594)
  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            "Unpublished"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-CHMP4B Fusion Vector
FEATURES             Location/Qualifiers
     source          1..5594
                     /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(1648..1666)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1649..1666)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     enhancer        1786..2089
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     2244..2264
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        2297..2344
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2376..3170
                     /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(2430..2449)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3040..3059
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(3328..3916)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3408..3427)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4042..4090
                     /note="synthetic polyadenylation signal"
     CDS             complement(4114..5151)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             4127..4135
                     /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(5182..5539)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5196..5331
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5348..5539
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5496..5516
                     /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 ttaaaccatg gatccagccc
      421 agttctccaa ttccttcatg tcgtcgtcct cctcttcttt cttcttggcg ggttttgatg
      481 gtagggctat agagggaaca tttggtagag ggactgtttc gggtccactg atttccagca
      541 aattcttgtc tagttcctcc tgttctagtt cttctaattc cgccatgagc tcatcctcgt
      601 caaactcttc tccaaaccct acaggtttcg aaattgctgt tgaaatctcc tctgcaagtt
      661 cttgctggtc agcaatgtcc tgcattaact catcaacttt atcgatgtcc atgttgtcat
      721 gggccgcctt catggccttg gcggcatagc ccatgttctt gagcacctcg gtgttggtgt
      781 tggcattctc cagggcctcc cgctggaact cgatggttga taatgtgccg tcgatctgcg
      841 ccagctgctt ctcatacctc ttcttacgct tcagtgcctg gagggccgcg cgcttgtttt
      901 tggtgccgtg cttcttggcg gccgtcagct cctgctcgat tttcttctcc aggaactcct
      961 gtttcttgct taacatctct tccgtgtccc gcagccgctg gatggcctcc tggggggtcg
     1021 ggccgccctt gccggcctta ccccctccag ccccgaacag cttcccgaac accgacatgg
     1081 cgatcgcacc gctcgagcct ccacctccgc tcccgccacc accggagctc ccactgttga
     1141 tggttactcg gaacagcatg gagccgtcgg gggtgatcag gcgctcgtcg ataattttgt
     1201 tgccgttcca cagggtccct gttacagtga tctttttgcc gtcgaacacg gcgatgcctt
     1261 catacggccg tccgaaatag ttcagcatgt tcggcgtaac cccgtcgatt accagtgtgc
     1321 catagggcag gatcacctta aagtgatgat catccacagg gtacaccacc ttaaacacct
     1381 cttcgatctg ggccatttgg tcggcgctca gaccttcata cgggatgatg acatggatgt
     1441 cgatcttcag ggcattttca ccgctccgga caatcctttg gatcggagtt acggacacgg
     1501 cgagattctg cagcaaactg gacacacctc cctgttcaag gacttggtcc aggttgtagg
     1561 cggctgtctg ttcccagtcc ccaacgaaat cttcgagtgt gaagaccatg gtgagcgcta
     1621 gctgcaaaaa gaacaagtag cttgtattct atagtgtcac ctaaatgggc gatctgacgg
     1681 ttcactaaac cagctctgct tatataggcc tccctggcgc gccactagtt attaatagta
     1741 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
     1801 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
     1861 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
     1921 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagtc cgccccctat
     1981 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttacggga
     2041 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
     2101 ttggcagtac accaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
     2161 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
     2221 tcgtaataac cccgccccgt tgacgcaaat gggcggtagg cgtgtacggt gggaggtcaa
     2281 atcgaattct aatactgttg acaattaatc atcggcatag tatatcggca tagtataata
     2341 cgacaaggtg aggaactaaa cccaggaggc agatcatgat tgaacaagat ggattgcacg
     2401 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     2461 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     2521 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt
     2581 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     2641 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     2701 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     2761 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     2821 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     2881 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg
     2941 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     3001 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     3061 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     3121 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga ttcgaaatga
     3181 ccgaccaagc gacgcccaac cggtatcagc tcactcaaag gcggtaatac ggttatccac
     3241 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     3301 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     3361 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     3421 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     3481 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     3541 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     3601 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     3661 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     3721 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     3781 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     3841 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     3901 aaaaaaagga tttcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     3961 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     4021 ccttttatag tccggaatcg tcacacaaaa aaccaacaca cagatgtaat gaaaataaag
     4081 atattttatt gcggccatcg tgatggctag agtctaaact cgacctacct ccttagcgcg
     4141 cggtcgtgta ctgggccggc ggttgccgct gtcggccagc acctcgacgc agccgtcggt
     4201 ccatacggct gcgctccggc gagcgatttg tgtgcgaccg acggtgccgg ccccgctgcg
     4261 gacgatggca tcgcagcggc cttgagccca ggcagcatcg tcgaagttgc cgtccacgag
     4321 gctctggtag agctgatcca ggccgatgcg cagcatgtag gctcgcagac gagggctgcc
     4381 ggccagctcg ggatgccggc gctcgaagta gcgagtctgc tgctccatgc aagccagcca
     4441 gggccgccag aagaagatgt tggccacctc gtactgactg tccccgaaca tagcttcgga
     4501 ccagtcgatt acggcggtga tgcggccgtt gtcggtcagg acgttgttgc tgccgaagtc
     4561 ggcatggacg aggtggcgca cctcgggaca gtcttcggcc cacagcatca gttcgtccag
     4621 ggcttgagct acgctggcgg acacggtgtc gtccatcacg gtctgccagt ggtagacatg
     4681 gggatcagca atggcgcaaa tgaaatcccg ccaggttgtg tactggccga tgccttgggg
     4741 cccgaatggg ccgaagccgc tggtttgact gaggtcggcg gcggcaatag catccatagc
     4801 ttcggcgaca ggctgtaaca cagcaggcag ctctgtttcg gggaggtctt ggagagtaac
     4861 gccttgggcg cgtctactga tgcagtatgt caggctctcg ctgaactcgc cgatgtccaa
     4921 cacttcgggg atgggtagtg cagcgctggc gaagtggcgg tacacgtagc ggtctttgta
     4981 gaagccatca gcgcagctat tcacccgcag tacatagccg cgtccgccga catcgaagct
     5041 gaaggctcgg ctctcttcgc cctccgacaa ctgcatcagg tcgctcacac tgtcgaactt
     5101 ctcgatgaga aatttttcaa cgctggtagc ggtgagttcg ggcttcttca tggtggcgct
     5161 agtgtcagaa gaatcgagct ttttgcaaaa gcctaggcct ccaaaaaagc ctcctcacta
     5221 cttctggaat agctcagagg ccgaggcggc ctcggcctct gcataaataa aaaaaattag
     5281 tcagccatgg ggcggagaat gggcggaact gggcggagtt aggggcggga tgggcggagt
     5341 taggggcggg actatggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc
     5401 tggggagcct ggggactttc cacacctggt tgctgactaa ttgagatgca tgctttgcat
     5461 acttctgcct gctggggagc ctggggactt tccacaccct aactgacaca cattccacag
     5521 ctggttcttt ccgcctcaga aggtacctaa ccaagttcct ctttcagagg ttatttcagg
     5581 ccatggtgct gcgc
//
