LOCUS       sequence-471904-        5780 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express PPP1CA-LgBiT 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 5780)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5780)
  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            "Unpublished"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: PPP1CA-LgBiT Fusion Vector
FEATURES             Location/Qualifiers
     source          1..5780
                     /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"
     promoter        complement(1834..1852)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1835..1852)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     enhancer        1972..2275
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     2430..2450
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        2483..2530
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2562..3356
                     /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(2616..2635)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3226..3245
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(3514..4102)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3594..3613)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4228..4276
                     /note="synthetic polyadenylation signal"
     CDS             complement(4300..5337)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             4313..4321
                     /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(5368..5725)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5382..5517
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5534..5725
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5682..5702
                     /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 ctagagtcgc ggccttagct gttgatggtt actcggaaca
      301 gcatggagcc gtcgggggtg atcaggcgct cgtcgataat tttgttgccg ttccacaggg
      361 tccctgttac agtgatcttt ttgccgtcga acacggcgat gccttcatac ggccgtccga
      421 aatagttcag catgttcggc gtaaccccgt cgattaccag tgtgccatag ggcaggatca
      481 ccttaaagtg atgatcatcc acagggtaca ccaccttaaa cacctcttcg atctgggcca
      541 tttggtcggc gctcagacct tcatacggga tgatgacatg gatgtcgatc ttcagggcat
      601 tttcaccgct ccggacaatc ctttggatcg gagttacgga cacggcgaga ttctgcagca
      661 aactggacac acctccctgt tcaaggactt ggtccaggtt gtaggcggct gtctgttccc
      721 agtccccaac gaaatcttcg agtgtgaaga caccgctcga gcctccacct ccgctcccgc
      781 caccacctga actgccttga gaaactttct tggctttggc ggaattgcgg ggtggggtga
      841 tgggtcggcc tccagggttc aggccactga actgcccgta cttccccttg ttcttgtcgg
      901 cgggcttgag gatctggaaa gagcacatga gggtctcgtc cacactcatc atggcgccag
      961 cattgtcaaa ctcgccacag tagttgggag ctgagaaaag tgtcaccagc tgccgcttgg
     1021 caaagaactc gtagccgtct tctaccacct ggtgtgctcg gcagatgagg tccaagtcgt
     1081 gcttgtggag gaacttggcc accacctcgg ctccaaaggt aaaagagacg ccacggtcgt
     1141 tctcgcccca gccctgcacg tccttgtcag ggtcagacca cagcaggtca cacagcaggc
     1201 cctggtcagg cacatctgtg ggccgcatga tccgccgaat ctgctccata gactgcaggt
     1261 ccggggacag gcctccgtgg cagcagaaga tcttttcgtc cactatggcc gcgatgggca
     1321 ggcagttgaa gcagtcagtg aaggttttcc acagtttgat gttgtagcgt ctcttgcact
     1381 catcgtagaa accatagatg cggttgatgc tggcacactc gtggttccca cggagcagga
     1441 agaagttctc ggggtacttg atcttatagg ccagcagcag gcagatggtc tccaaggact
     1501 gcttgcccct gtccacatag tcccccagaa agaggtagtt gctctcggga gggaaaccgc
     1561 catactcaaa tagtcgcaga aggtcgtagt actggccgtg tatgtcaccg cagatcttga
     1621 ggggtgcctc cagctccaga agaatgggct ggctcagaaa aatctcccgg gatttcaggc
     1681 acagaccgcg gatctcgttc tctgtcagct gtacattctt gccaggccgc gagccctgca
     1741 cttccagcag gcgcccgatg atcgagtcca ggttgagctt ctcgctgtcg gacatggcga
     1801 tcgctagctg caaaaagaac aagtagcttg tattctatag tgtcacctaa atgggcgatc
     1861 tgacggttca ctaaaccagc tctgcttata taggcctccc tggcgcgcca ctagttatta
     1921 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
     1981 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
     2041 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
     2101 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtccgcc
     2161 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
     2221 acgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
     2281 gcggttttgg cagtacacca atgggcgtgg atagcggttt gactcacggg gatttccaag
     2341 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
     2401 aaaatgtcgt aataaccccg ccccgttgac gcaaatgggc ggtaggcgtg tacggtggga
     2461 ggtcaaatcg aattctaata ctgttgacaa ttaatcatcg gcatagtata tcggcatagt
     2521 ataatacgac aaggtgagga actaaaccca ggaggcagat catgattgaa caagatggat
     2581 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     2641 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     2701 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc
     2761 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     2821 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     2881 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     2941 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     3001 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     3061 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga
     3121 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     3181 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     3241 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     3301 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgattcg
     3361 aaatgaccga ccaagcgacg cccaaccggt atcagctcac tcaaaggcgg taatacggtt
     3421 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     3481 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     3541 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     3601 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     3661 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     3721 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     3781 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     3841 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     3901 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     3961 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     4021 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     4081 gcgcagaaaa aaaggatttc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     4141 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     4201 ctagatcctt ttatagtccg gaatcgtcac acaaaaaacc aacacacaga tgtaatgaaa
     4261 ataaagatat tttattgcgg ccatcgtgat ggctagagtc taaactcgac ctacctcctt
     4321 agcgcgcggt cgtgtactgg gccggcggtt gccgctgtcg gccagcacct cgacgcagcc
     4381 gtcggtccat acggctgcgc tccggcgagc gatttgtgtg cgaccgacgg tgccggcccc
     4441 gctgcggacg atggcatcgc agcggccttg agcccaggca gcatcgtcga agttgccgtc
     4501 cacgaggctc tggtagagct gatccaggcc gatgcgcagc atgtaggctc gcagacgagg
     4561 gctgccggcc agctcgggat gccggcgctc gaagtagcga gtctgctgct ccatgcaagc
     4621 cagccagggc cgccagaaga agatgttggc cacctcgtac tgactgtccc cgaacatagc
     4681 ttcggaccag tcgattacgg cggtgatgcg gccgttgtcg gtcaggacgt tgttgctgcc
     4741 gaagtcggca tggacgaggt ggcgcacctc gggacagtct tcggcccaca gcatcagttc
     4801 gtccagggct tgagctacgc tggcggacac ggtgtcgtcc atcacggtct gccagtggta
     4861 gacatgggga tcagcaatgg cgcaaatgaa atcccgccag gttgtgtact ggccgatgcc
     4921 ttggggcccg aatgggccga agccgctggt ttgactgagg tcggcggcgg caatagcatc
     4981 catagcttcg gcgacaggct gtaacacagc aggcagctct gtttcgggga ggtcttggag
     5041 agtaacgcct tgggcgcgtc tactgatgca gtatgtcagg ctctcgctga actcgccgat
     5101 gtccaacact tcggggatgg gtagtgcagc gctggcgaag tggcggtaca cgtagcggtc
     5161 tttgtagaag ccatcagcgc agctattcac ccgcagtaca tagccgcgtc cgccgacatc
     5221 gaagctgaag gctcggctct cttcgccctc cgacaactgc atcaggtcgc tcacactgtc
     5281 gaacttctcg atgagaaatt tttcaacgct ggtagcggtg agttcgggct tcttcatggt
     5341 ggcgctagtg tcagaagaat cgagcttttt gcaaaagcct aggcctccaa aaaagcctcc
     5401 tcactacttc tggaatagct cagaggccga ggcggcctcg gcctctgcat aaataaaaaa
     5461 aattagtcag ccatggggcg gagaatgggc ggaactgggc ggagttaggg gcgggatggg
     5521 cggagttagg ggcgggacta tggttgctga ctaattgaga tgcatgcttt gcatacttct
     5581 gcctgctggg gagcctgggg actttccaca cctggttgct gactaattga gatgcatgct
     5641 ttgcatactt ctgcctgctg gggagcctgg ggactttcca caccctaact gacacacatt
     5701 ccacagctgg ttctttccgc ctcagaaggt acctaaccaa gttcctcttt cagaggttat
     5761 ttcaggccat ggtgctgcgc
//
