LOCUS       sequence-470641-        5912 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express LgBiT-PPP1CA 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 5912)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5912)
  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            "Unpublished"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-PPP1CA Fusion Vector
FEATURES             Location/Qualifiers
     source          1..5912
                     /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(1966..1984)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1967..1984)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     enhancer        2104..2407
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     2562..2582
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        2615..2662
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2694..3488
                     /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(2748..2767)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3358..3377
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(3646..4234)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3726..3745)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4360..4408
                     /note="synthetic polyadenylation signal"
     CDS             complement(4432..5469)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             4445..4453
                     /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(5500..5857)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5514..5649
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5666..5857
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5814..5834
                     /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 ttaaactttc ttggctttgg
      421 cggaattgcg gggtggggtg atgggtcggc ctccagggtt caggccactg aactgcccgt
      481 acttcccctt gttcttgtcg gcgggcttga ggatctggaa agagcacatg agggtctcgt
      541 ccacactcat catggcgcca gcattgtcaa actcgccaca gtagttggga gctgagaaaa
      601 gtgtcaccag ctgccgcttg gcaaagaact cgtagccgtc ttctaccacc tggtgtgctc
      661 ggcagatgag gtccaagtcg tgcttgtgga ggaacttggc caccacctcg gctccaaagg
      721 taaaagagac gccacggtcg ttctcgcccc agccctgcac gtccttgtca gggtcagacc
      781 acagcaggtc acacagcagg ccctggtcag gcacatctgt gggccgcatg atccgccgaa
      841 tctgctccat agactgcagg tccggggaca ggcctccgtg gcagcagaag atcttttcgt
      901 ccactatggc cgcgatgggc aggcagttga agcagtcagt gaaggttttc cacagtttga
      961 tgttgtagcg tctcttgcac tcatcgtaga aaccatagat gcggttgatg ctggcacact
     1021 cgtggttccc acggagcagg aagaagttct cggggtactt gatcttatag gccagcagca
     1081 ggcagatggt ctccaaggac tgcttgcccc tgtccacata gtcccccaga aagaggtagt
     1141 tgctctcggg agggaaaccg ccatactcaa atagtcgcag aaggtcgtag tactggccgt
     1201 gtatgtcacc gcagatcttg aggggtgcct ccagctccag aagaatgggc tggctcagaa
     1261 aaatctcccg ggatttcagg cacagaccgc ggatctcgtt ctctgtcagc tgtacattct
     1321 tgccaggccg cgagccctgc acttccagca ggcgcccgat gatcgagtcc aggttgagct
     1381 tctcgctgtc ggacatggcg atcgcaccgc tcgagcctcc acctccgctc ccgccaccac
     1441 cggagctccc actgttgatg gttactcgga acagcatgga gccgtcgggg gtgatcaggc
     1501 gctcgtcgat aattttgttg ccgttccaca gggtccctgt tacagtgatc tttttgccgt
     1561 cgaacacggc gatgccttca tacggccgtc cgaaatagtt cagcatgttc ggcgtaaccc
     1621 cgtcgattac cagtgtgcca tagggcagga tcaccttaaa gtgatgatca tccacagggt
     1681 acaccacctt aaacacctct tcgatctggg ccatttggtc ggcgctcaga ccttcatacg
     1741 ggatgatgac atggatgtcg atcttcaggg cattttcacc gctccggaca atcctttgga
     1801 tcggagttac ggacacggcg agattctgca gcaaactgga cacacctccc tgttcaagga
     1861 cttggtccag gttgtaggcg gctgtctgtt cccagtcccc aacgaaatct tcgagtgtga
     1921 agaccatggt gagcgctagc tgcaaaaaga acaagtagct tgtattctat agtgtcacct
     1981 aaatgggcga tctgacggtt cactaaacca gctctgctta tataggcctc cctggcgcgc
     2041 cactagttat taatagtaat caattacggg gtcattagtt catagcccat atatggagtt
     2101 ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
     2161 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
     2221 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
     2281 gccaagtccg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
     2341 gtacatgacc ttacgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
     2401 taccatggtg atgcggtttt ggcagtacac caatgggcgt ggatagcggt ttgactcacg
     2461 gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca
     2521 acgggacttt ccaaaatgtc gtaataaccc cgccccgttg acgcaaatgg gcggtaggcg
     2581 tgtacggtgg gaggtcaaat cgaattctaa tactgttgac aattaatcat cggcatagta
     2641 tatcggcata gtataatacg acaaggtgag gaactaaacc caggaggcag atcatgattg
     2701 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     2761 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     2821 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg
     2881 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     2941 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     3001 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     3061 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     3121 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     3181 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg
     3241 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     3301 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     3361 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     3421 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     3481 tcttctgatt cgaaatgacc gaccaagcga cgcccaaccg gtatcagctc actcaaaggc
     3541 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     3601 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     3661 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     3721 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     3781 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     3841 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     3901 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     3961 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     4021 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     4081 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     4141 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     4201 gcagcagatt acgcgcagaa aaaaaggatt tcaagaagat cctttgatct tttctacggg
     4261 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     4321 aaggatcttc acctagatcc ttttatagtc cggaatcgtc acacaaaaaa ccaacacaca
     4381 gatgtaatga aaataaagat attttattgc ggccatcgtg atggctagag tctaaactcg
     4441 acctacctcc ttagcgcgcg gtcgtgtact gggccggcgg ttgccgctgt cggccagcac
     4501 ctcgacgcag ccgtcggtcc atacggctgc gctccggcga gcgatttgtg tgcgaccgac
     4561 ggtgccggcc ccgctgcgga cgatggcatc gcagcggcct tgagcccagg cagcatcgtc
     4621 gaagttgccg tccacgaggc tctggtagag ctgatccagg ccgatgcgca gcatgtaggc
     4681 tcgcagacga gggctgccgg ccagctcggg atgccggcgc tcgaagtagc gagtctgctg
     4741 ctccatgcaa gccagccagg gccgccagaa gaagatgttg gccacctcgt actgactgtc
     4801 cccgaacata gcttcggacc agtcgattac ggcggtgatg cggccgttgt cggtcaggac
     4861 gttgttgctg ccgaagtcgg catggacgag gtggcgcacc tcgggacagt cttcggccca
     4921 cagcatcagt tcgtccaggg cttgagctac gctggcggac acggtgtcgt ccatcacggt
     4981 ctgccagtgg tagacatggg gatcagcaat ggcgcaaatg aaatcccgcc aggttgtgta
     5041 ctggccgatg ccttggggcc cgaatgggcc gaagccgctg gtttgactga ggtcggcggc
     5101 ggcaatagca tccatagctt cggcgacagg ctgtaacaca gcaggcagct ctgtttcggg
     5161 gaggtcttgg agagtaacgc cttgggcgcg tctactgatg cagtatgtca ggctctcgct
     5221 gaactcgccg atgtccaaca cttcggggat gggtagtgca gcgctggcga agtggcggta
     5281 cacgtagcgg tctttgtaga agccatcagc gcagctattc acccgcagta catagccgcg
     5341 tccgccgaca tcgaagctga aggctcggct ctcttcgccc tccgacaact gcatcaggtc
     5401 gctcacactg tcgaacttct cgatgagaaa tttttcaacg ctggtagcgg tgagttcggg
     5461 cttcttcatg gtggcgctag tgtcagaaga atcgagcttt ttgcaaaagc ctaggcctcc
     5521 aaaaaagcct cctcactact tctggaatag ctcagaggcc gaggcggcct cggcctctgc
     5581 ataaataaaa aaaattagtc agccatgggg cggagaatgg gcggaactgg gcggagttag
     5641 gggcgggatg ggcggagtta ggggcgggac tatggttgct gactaattga gatgcatgct
     5701 ttgcatactt ctgcctgctg gggagcctgg ggactttcca cacctggttg ctgactaatt
     5761 gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc cacaccctaa
     5821 ctgacacaca ttccacagct ggttctttcc gcctcagaag gtacctaacc aagttcctct
     5881 ttcagaggtt atttcaggcc atggtgctgc gc
//
