LOCUS       sequence-479834-        5447 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Express LgBiT-BCL2A1 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 5447)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5447)
  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            "Unpublished"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-BCL2A1 Vector
FEATURES             Location/Qualifiers
     source          1..5447
                     /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(1501..1519)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1502..1519)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     enhancer        1639..1942
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     2097..2117
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        2150..2197
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2229..3023
                     /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(2283..2302)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2893..2912
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(3181..3769)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3261..3280)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3895..3943
                     /note="synthetic polyadenylation signal"
     CDS             complement(3967..5004)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             3980..3988
                     /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(5035..5392)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5049..5184
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5201..5392
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5349..5369
                     /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 ttaaacacag tattgcttca
      421 ggagagatag catttcacag atctttcctg taacttctag aaaagtcatc cagccagatt
      481 taggttcaaa cttctttaca aagccatttt cccagcctcc gttttgcctt atccattctc
      541 ctgtgttatt cattatgaac tccgcaacaa aatatgaaat ctccttatag gtatccacat
      601 ccggggcaat ttgctgtcgt agaagtttct tgatgagaat accttcaaat gcaaatatgg
      661 ttacaattct tccccagtta atgatgccgt cttcaaactc cttttccatc acttggttga
      721 atagtgttct ggcagtgtct acggacacaa cattaacatt gtccaagcat gacttcagat
      781 tcttttccac ttctttttgg actgagaacg caacattttg tagcactctg gacgttttgc
      841 ttggacctga tccaggttgt ggtatctgta ggacgcactg cagatagtcc tgagccagcc
      901 tgtaaatata tccaaattca cagtctgtca tggcgatcgc accgctcgag cctccacctc
      961 cgctcccgcc accaccggag ctcccactgt tgatggttac tcggaacagc atggagccgt
     1021 cgggggtgat caggcgctcg tcgataattt tgttgccgtt ccacagggtc cctgttacag
     1081 tgatcttttt gccgtcgaac acggcgatgc cttcatacgg ccgtccgaaa tagttcagca
     1141 tgttcggcgt aaccccgtcg attaccagtg tgccataggg caggatcacc ttaaagtgat
     1201 gatcatccac agggtacacc accttaaaca cctcttcgat ctgggccatt tggtcggcgc
     1261 tcagaccttc atacgggatg atgacatgga tgtcgatctt cagggcattt tcaccgctcc
     1321 ggacaatcct ttggatcgga gttacggaca cggcgagatt ctgcagcaaa ctggacacac
     1381 ctccctgttc aaggacttgg tccaggttgt aggcggctgt ctgttcccag tccccaacga
     1441 aatcttcgag tgtgaagacc atggtgagcg ctagctgcaa aaagaacaag tagcttgtat
     1501 tctatagtgt cacctaaatg ggcgatctga cggttcacta aaccagctct gcttatatag
     1561 gcctccctgg cgcgccacta gttattaata gtaatcaatt acggggtcat tagttcatag
     1621 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
     1681 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
     1741 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
     1801 tcaagtgtat catatgccaa gtccgccccc tattgacgtc aatgacggta aatggcccgc
     1861 ctggcattat gcccagtaca tgaccttacg ggactttcct acttggcagt acatctacgt
     1921 attagtcatc gctattacca tggtgatgcg gttttggcag tacaccaatg ggcgtggata
     1981 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
     2041 ttggcaccaa aatcaacggg actttccaaa atgtcgtaat aaccccgccc cgttgacgca
     2101 aatgggcggt aggcgtgtac ggtgggaggt caaatcgaat tctaatactg ttgacaatta
     2161 atcatcggca tagtatatcg gcatagtata atacgacaag gtgaggaact aaacccagga
     2221 ggcagatcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     2281 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     2341 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     2401 atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     2461 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     2521 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     2581 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     2641 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     2701 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca
     2761 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     2821 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     2881 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     2941 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     3001 gccttcttga cgagttcttc tgattcgaaa tgaccgacca agcgacgccc aaccggtatc
     3061 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     3121 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     3181 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     3241 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     3301 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     3361 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     3421 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     3481 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     3541 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     3601 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     3661 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     3721 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatttcaag aagatccttt
     3781 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     3841 catgagatta tcaaaaagga tcttcaccta gatcctttta tagtccggaa tcgtcacaca
     3901 aaaaaccaac acacagatgt aatgaaaata aagatatttt attgcggcca tcgtgatggc
     3961 tagagtctaa actcgaccta cctccttagc gcgcggtcgt gtactgggcc ggcggttgcc
     4021 gctgtcggcc agcacctcga cgcagccgtc ggtccatacg gctgcgctcc ggcgagcgat
     4081 ttgtgtgcga ccgacggtgc cggccccgct gcggacgatg gcatcgcagc ggccttgagc
     4141 ccaggcagca tcgtcgaagt tgccgtccac gaggctctgg tagagctgat ccaggccgat
     4201 gcgcagcatg taggctcgca gacgagggct gccggccagc tcgggatgcc ggcgctcgaa
     4261 gtagcgagtc tgctgctcca tgcaagccag ccagggccgc cagaagaaga tgttggccac
     4321 ctcgtactga ctgtccccga acatagcttc ggaccagtcg attacggcgg tgatgcggcc
     4381 gttgtcggtc aggacgttgt tgctgccgaa gtcggcatgg acgaggtggc gcacctcggg
     4441 acagtcttcg gcccacagca tcagttcgtc cagggcttga gctacgctgg cggacacggt
     4501 gtcgtccatc acggtctgcc agtggtagac atggggatca gcaatggcgc aaatgaaatc
     4561 ccgccaggtt gtgtactggc cgatgccttg gggcccgaat gggccgaagc cgctggtttg
     4621 actgaggtcg gcggcggcaa tagcatccat agcttcggcg acaggctgta acacagcagg
     4681 cagctctgtt tcggggaggt cttggagagt aacgccttgg gcgcgtctac tgatgcagta
     4741 tgtcaggctc tcgctgaact cgccgatgtc caacacttcg gggatgggta gtgcagcgct
     4801 ggcgaagtgg cggtacacgt agcggtcttt gtagaagcca tcagcgcagc tattcacccg
     4861 cagtacatag ccgcgtccgc cgacatcgaa gctgaaggct cggctctctt cgccctccga
     4921 caactgcatc aggtcgctca cactgtcgaa cttctcgatg agaaattttt caacgctggt
     4981 agcggtgagt tcgggcttct tcatggtggc gctagtgtca gaagaatcga gctttttgca
     5041 aaagcctagg cctccaaaaa agcctcctca ctacttctgg aatagctcag aggccgaggc
     5101 ggcctcggcc tctgcataaa taaaaaaaat tagtcagcca tggggcggag aatgggcgga
     5161 actgggcgga gttaggggcg ggatgggcgg agttaggggc gggactatgg ttgctgacta
     5221 attgagatgc atgctttgca tacttctgcc tgctggggag cctggggact ttccacacct
     5281 ggttgctgac taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga
     5341 ctttccacac cctaactgac acacattcca cagctggttc tttccgcctc agaaggtacc
     5401 taaccaagtt cctctttcag aggttatttc aggccatggt gctgcgc
//
