LOCUS       sequence-479835-        5501 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Express LgBiT-BCL2L2 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 5501)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5501)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 7, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: LgBiT-BCL2L2 Vector
FEATURES             Location/Qualifiers
     source          1..5501
                     /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(1555..1573)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1556..1573)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     enhancer        1693..1996
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     2151..2171
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        2204..2251
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2283..3077
                     /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(2337..2356)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2947..2966
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(3235..3823)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3315..3334)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3949..3997
                     /note="synthetic polyadenylation signal"
     CDS             complement(4021..5058)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             4034..4042
                     /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(5089..5446)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5103..5238
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5255..5446
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5403..5423
                     /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 ttaaaccttg ctagcaaaaa
      421 aggcccctac agttaccagg gcccccagtg ccacggcccc cgtcagaact gtcctcactg
      481 atgcccagtt accctcccgc agacgccgcg cctcttccag agccccgtca ccgtatagag
      541 ctgtgaactc cgcccagccc ccactgctgt ggatccagtc agccagccgc gtctccaggt
      601 aggccaccat ccactcctgc acttgtccca ccagtggttc catctccttg ttgacactct
      661 cagcacacag tgcagcccca aagacaaaga aggctacaag gcggccccag ttggggcccc
      721 cttgaaaaag ttcatcggag acctgggtga agcgttgttg ggctgagcct ggggtcacat
      781 gcagctgagc cgccagatca gagaaggtgc gccggaagcg ggtctcgaac tcatctccag
      841 ctgcccgcat ggcttggtgc agcgggtcag ctgctgggcc ctccccgggg ccagctccac
      901 agacataacc cttctgcctc agcttataac ctacaaagtc tgccaccaga gcccgtgtgt
      961 ctggggccga ggctggggtc gccatggcga tcgcaccgct cgagcctcca cctccgctcc
     1021 cgccaccacc ggagctccca ctgttgatgg ttactcggaa cagcatggag ccgtcggggg
     1081 tgatcaggcg ctcgtcgata attttgttgc cgttccacag ggtccctgtt acagtgatct
     1141 ttttgccgtc gaacacggcg atgccttcat acggccgtcc gaaatagttc agcatgttcg
     1201 gcgtaacccc gtcgattacc agtgtgccat agggcaggat caccttaaag tgatgatcat
     1261 ccacagggta caccacctta aacacctctt cgatctgggc catttggtcg gcgctcagac
     1321 cttcatacgg gatgatgaca tggatgtcga tcttcagggc attttcaccg ctccggacaa
     1381 tcctttggat cggagttacg gacacggcga gattctgcag caaactggac acacctccct
     1441 gttcaaggac ttggtccagg ttgtaggcgg ctgtctgttc ccagtcccca acgaaatctt
     1501 cgagtgtgaa gaccatggtg agcgctagct gcaaaaagaa caagtagctt gtattctata
     1561 gtgtcaccta aatgggcgat ctgacggttc actaaaccag ctctgcttat ataggcctcc
     1621 ctggcgcgcc actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
     1681 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
     1741 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
     1801 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
     1861 gtatcatatg ccaagtccgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     1921 ttatgcccag tacatgacct tacgggactt tcctacttgg cagtacatct acgtattagt
     1981 catcgctatt accatggtga tgcggttttg gcagtacacc aatgggcgtg gatagcggtt
     2041 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
     2101 ccaaaatcaa cgggactttc caaaatgtcg taataacccc gccccgttga cgcaaatggg
     2161 cggtaggcgt gtacggtggg aggtcaaatc gaattctaat actgttgaca attaatcatc
     2221 ggcatagtat atcggcatag tataatacga caaggtgagg aactaaaccc aggaggcaga
     2281 tcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     2341 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     2401 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     2461 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     2521 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     2581 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     2641 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     2701 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     2761 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
     2821 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     2881 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     2941 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     3001 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     3061 ttgacgagtt cttctgattc gaaatgaccg accaagcgac gcccaaccgg tatcagctca
     3121 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     3181 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     3241 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     3301 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     3361 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     3421 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     3481 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     3541 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     3601 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     3661 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     3721 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     3781 tgtttgcaag cagcagatta cgcgcagaaa aaaaggattt caagaagatc ctttgatctt
     3841 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     3901 attatcaaaa aggatcttca cctagatcct tttatagtcc ggaatcgtca cacaaaaaac
     3961 caacacacag atgtaatgaa aataaagata ttttattgcg gccatcgtga tggctagagt
     4021 ctaaactcga cctacctcct tagcgcgcgg tcgtgtactg ggccggcggt tgccgctgtc
     4081 ggccagcacc tcgacgcagc cgtcggtcca tacggctgcg ctccggcgag cgatttgtgt
     4141 gcgaccgacg gtgccggccc cgctgcggac gatggcatcg cagcggcctt gagcccaggc
     4201 agcatcgtcg aagttgccgt ccacgaggct ctggtagagc tgatccaggc cgatgcgcag
     4261 catgtaggct cgcagacgag ggctgccggc cagctcggga tgccggcgct cgaagtagcg
     4321 agtctgctgc tccatgcaag ccagccaggg ccgccagaag aagatgttgg ccacctcgta
     4381 ctgactgtcc ccgaacatag cttcggacca gtcgattacg gcggtgatgc ggccgttgtc
     4441 ggtcaggacg ttgttgctgc cgaagtcggc atggacgagg tggcgcacct cgggacagtc
     4501 ttcggcccac agcatcagtt cgtccagggc ttgagctacg ctggcggaca cggtgtcgtc
     4561 catcacggtc tgccagtggt agacatgggg atcagcaatg gcgcaaatga aatcccgcca
     4621 ggttgtgtac tggccgatgc cttggggccc gaatgggccg aagccgctgg tttgactgag
     4681 gtcggcggcg gcaatagcat ccatagcttc ggcgacaggc tgtaacacag caggcagctc
     4741 tgtttcgggg aggtcttgga gagtaacgcc ttgggcgcgt ctactgatgc agtatgtcag
     4801 gctctcgctg aactcgccga tgtccaacac ttcggggatg ggtagtgcag cgctggcgaa
     4861 gtggcggtac acgtagcggt ctttgtagaa gccatcagcg cagctattca cccgcagtac
     4921 atagccgcgt ccgccgacat cgaagctgaa ggctcggctc tcttcgccct ccgacaactg
     4981 catcaggtcg ctcacactgt cgaacttctc gatgagaaat ttttcaacgc tggtagcggt
     5041 gagttcgggc ttcttcatgg tggcgctagt gtcagaagaa tcgagctttt tgcaaaagcc
     5101 taggcctcca aaaaagcctc ctcactactt ctggaatagc tcagaggccg aggcggcctc
     5161 ggcctctgca taaataaaaa aaattagtca gccatggggc ggagaatggg cggaactggg
     5221 cggagttagg ggcgggatgg gcggagttag gggcgggact atggttgctg actaattgag
     5281 atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac acctggttgc
     5341 tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc
     5401 acaccctaac tgacacacat tccacagctg gttctttccg cctcagaagg tacctaacca
     5461 agttcctctt tcagaggtta tttcaggcca tggtgctgcg c
//
