LOCUS       sequence-471905-        5292 bp    DNA     circular SYN 14-JUL-2025
DEFINITION  Express PPP1CA-SmBiT 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 5292)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 5292)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: PPP1CA-SmBiT Fusion Vector
FEATURES             Location/Qualifiers
     source          1..5292
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    24..105
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     enhancer        338..641
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        642..845
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     796..816
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          981..1113
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1096..1115
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1157..1176
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1157..1175
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    2355..2476
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2366..2385)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(2591..2624)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2634..2681
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2713..3507
                     /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(2767..2786)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3377..3396
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(3665..4253)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3745..3764)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4379..4427
                     /note="synthetic polyadenylation signal"
     CDS             complement(4451..4849)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      4846..4855
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4880..5237)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4894..5029
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5046..5237
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5194..5214
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 aagacgcgtt aattaagacg tcggcagtga aaaaaatgct ttatttgtga aatttgtgat
       61 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
      121 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac
      181 ctctacaaat gtggtatggc tgattatgat ccgcggccgc aatacgtcga cgttatcagc
      241 tgacttcgta cgagagccta ggattatggc gcgccactag ttattaatag taatcaatta
      301 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      361 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      421 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      481 ctgcccactt ggcagtacat caagtgtatc atatgccaag tccgccccct attgacgtca
      541 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttacgg gactttccta
      601 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      661 acaccaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      721 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaata
      781 accccgcccc gttgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      841 gagctggttt agtgaaccgt cagatcacta gaagctttat tgcggtagtt tatcacagtt
      901 aaattgctaa cgcagtcagt gcttctgaca caacagtctc gaacttaagc tgcagaagtt
      961 ggtcgtgagg cactgggcag gtaagtatca aggttacaag acaggtttaa ggagaccaat
     1021 agaaactggg cttgtcgaga cagagaagac tcttgcgttt ctgataggca cctattggtc
     1081 ttactgacat ccactttgcc tttctctcca caggtgtcca ctcccagttc aattacagct
     1141 cttaaggcta gagtattaat acgactcact atagggctag cgatcgccat gtccgacagc
     1201 gagaagctca acctggactc gatcatcggg cgcctgctgg aagtgcaggg ctcgcggcct
     1261 ggcaagaatg tacagctgac agagaacgag atccgcggtc tgtgcctgaa atcccgggag
     1321 atttttctga gccagcccat tcttctggag ctggaggcac ccctcaagat ctgcggtgac
     1381 atacacggcc agtactacga ccttctgcga ctatttgagt atggcggttt ccctcccgag
     1441 agcaactacc tctttctggg ggactatgtg gacaggggca agcagtcctt ggagaccatc
     1501 tgcctgctgc tggcctataa gatcaagtac cccgagaact tcttcctgct ccgtgggaac
     1561 cacgagtgtg ccagcatcaa ccgcatctat ggtttctacg atgagtgcaa gagacgctac
     1621 aacatcaaac tgtggaaaac cttcactgac tgcttcaact gcctgcccat cgcggccata
     1681 gtggacgaaa agatcttctg ctgccacgga ggcctgtccc cggacctgca gtctatggag
     1741 cagattcggc ggatcatgcg gcccacagat gtgcctgacc agggcctgct gtgtgacctg
     1801 ctgtggtctg accctgacaa ggacgtgcag ggctggggcg agaacgaccg tggcgtctct
     1861 tttacctttg gagccgaggt ggtggccaag ttcctccaca agcacgactt ggacctcatc
     1921 tgccgagcac accaggtggt agaagacggc tacgagttct ttgccaagcg gcagctggtg
     1981 acacttttct cagctcccaa ctactgtggc gagtttgaca atgctggcgc catgatgagt
     2041 gtggacgaga ccctcatgtg ctctttccag atcctcaagc ccgccgacaa gaacaagggg
     2101 aagtacgggc agttcagtgg cctgaaccct ggaggccgac ccatcacccc accccgcaat
     2161 tccgccaaag ccaagaaagt ttctcaaggc agttcaggtg gtggcgggag cggaggtgga
     2221 ggctcgagcg gtgtgaccgg ctaccggctg ttcgaggaga ttctgtaagg ccgcgactct
     2281 agagtcgacc tgcaggcatg caagctgatt tggctgctgc caccgctgag caaggccgct
     2341 tcgagcagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     2401 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     2461 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga
     2521 gatgtgggag gtttttttaa gcaagtaaaa cctctacaaa tgtggtaaaa tcttaattaa
     2581 gaattctaat ataacttcgt atagcataca ttatacgaag ttatggatcc actgttgaca
     2641 attaatcatc ggcatagtat atcggcatag tataatacga caaggtgagg aactaaaccc
     2701 aggaggcaga tcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     2761 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     2821 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     2881 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     2941 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     3001 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     3061 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     3121 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     3181 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     3241 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     3301 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     3361 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     3421 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     3481 tatcgccttc ttgacgagtt cttctgattc gaaatgaccg accaagcgac gcccaaccgg
     3541 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     3601 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     3661 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     3721 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     3781 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     3841 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     3901 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     3961 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     4021 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     4081 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     4141 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     4201 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggattt caagaagatc
     4261 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     4321 tggtcatgag attatcaaaa aggatcttca cctagatcct tttatagtcc ggaatcgtca
     4381 cacaaaaaac caacacacag atgtaatgaa aataaagata ttttattgcg gccatcgtga
     4441 tggctagagt ttagccctcc cacacataac cagagggcag caattcacga atcccaactg
     4501 ccgtcggctg tccatcactg tccttcacta tggctttgat cccaggatgc agatcgagaa
     4561 gcacctgtcg gcaccgtccg caggggctca agatgcccct gttctcattt ccgatcgcga
     4621 cgatacaagt caggttgcca gctgccgcag cagcagcagt gcccagcacc acgagttctg
     4681 cacaaggtcc cccagtaaaa tgatatacat tgacaccagt gaagatgcgg ccgtcgctag
     4741 agagagctgc gctggcgacg ctgtagtctt cagagatggg gatgctgttg attgtagccg
     4801 ttgctctttc aatgagggtg gattcttctt gagacaaagg cttggccatg gtggcgctag
     4861 tgtcagaaga atcgagcttt ttgcaaaagc ctaggcctcc aaaaaagcct cctcactact
     4921 tctggaatag ctcagaggcc gaggcggcct cggcctctgc ataaataaaa aaaattagtc
     4981 agccatgggg cggagaatgg gcggaactgg gcggagttag gggcgggatg ggcggagtta
     5041 ggggcgggac tatggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg
     5101 gggagcctgg ggactttcca cacctggttg ctgactaatt gagatgcatg ctttgcatac
     5161 ttctgcctgc tggggagcct ggggactttc cacaccctaa ctgacacaca ttccacagct
     5221 ggttctttcc gcctcagaag gtacctaacc aagttcctct ttcagaggtt atttcaggcc
     5281 atggtgctgc gc
//
