LOCUS       sequence-471906-        5351 bp    DNA     circular SYN 14-JUL-2025
DEFINITION  Express SmBiT-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 5351)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 5351)
  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: SmBiT-PPP1CA Fusion Vector
FEATURES             Location/Qualifiers
     source          1..5351
                     /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    2414..2535
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2425..2444)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(2650..2683)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2693..2740
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2772..3566
                     /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(2826..2845)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3436..3455
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(3724..4312)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3804..3823)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4438..4486
                     /note="synthetic polyadenylation signal"
     CDS             complement(4510..4908)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      4905..4914
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4939..5296)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4953..5088
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5105..5296
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5253..5273
                     /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 cgctcaccat ggtgaccggc
     1201 taccggctgt tcgaggagat tctcgggagc tccggtggtg gcgggagcgg aggtggaggc
     1261 tcgagcggtg cgatcgccat gtccgacagc gagaagctca acctggactc gatcatcggg
     1321 cgcctgctgg aagtgcaggg ctcgcggcct ggcaagaatg tacagctgac agagaacgag
     1381 atccgcggtc tgtgcctgaa atcccgggag atttttctga gccagcccat tcttctggag
     1441 ctggaggcac ccctcaagat ctgcggtgac atacacggcc agtactacga ccttctgcga
     1501 ctatttgagt atggcggttt ccctcccgag agcaactacc tctttctggg ggactatgtg
     1561 gacaggggca agcagtcctt ggagaccatc tgcctgctgc tggcctataa gatcaagtac
     1621 cccgagaact tcttcctgct ccgtgggaac cacgagtgtg ccagcatcaa ccgcatctat
     1681 ggtttctacg atgagtgcaa gagacgctac aacatcaaac tgtggaaaac cttcactgac
     1741 tgcttcaact gcctgcccat cgcggccata gtggacgaaa agatcttctg ctgccacgga
     1801 ggcctgtccc cggacctgca gtctatggag cagattcggc ggatcatgcg gcccacagat
     1861 gtgcctgacc agggcctgct gtgtgacctg ctgtggtctg accctgacaa ggacgtgcag
     1921 ggctggggcg agaacgaccg tggcgtctct tttacctttg gagccgaggt ggtggccaag
     1981 ttcctccaca agcacgactt ggacctcatc tgccgagcac accaggtggt agaagacggc
     2041 tacgagttct ttgccaagcg gcagctggtg acacttttct cagctcccaa ctactgtggc
     2101 gagtttgaca atgctggcgc catgatgagt gtggacgaga ccctcatgtg ctctttccag
     2161 atcctcaagc ccgccgacaa gaacaagggg aagtacgggc agttcagtgg cctgaaccct
     2221 ggaggccgac ccatcacccc accccgcaat tccgccaaag ccaagaaagt ttaaacgaat
     2281 tcgggctcgg tacccgggga tcctctagag tcgacctgca ggcatgcaag ctgatccggc
     2341 tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc aaggccgctt
     2401 cgagcagaca tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga
     2461 aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc
     2521 tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag
     2581 atgtgggagg tttttttaag caagtaaaac ctctacaaat gtggtaaaat cttaattaag
     2641 aattctaata taacttcgta tagcatacat tatacgaagt tatggatcca ctgttgacaa
     2701 ttaatcatcg gcatagtata tcggcatagt ataatacgac aaggtgagga actaaaccca
     2761 ggaggcagat catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     2821 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     2881 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     2941 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     3001 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     3061 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     3121 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     3181 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     3241 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     3301 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     3361 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     3421 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     3481 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     3541 atcgccttct tgacgagttc ttctgattcg aaatgaccga ccaagcgacg cccaaccggt
     3601 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     3661 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     3721 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     3781 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     3841 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     3901 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     3961 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     4021 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     4081 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     4141 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     4201 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     4261 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatttc aagaagatcc
     4321 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     4381 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttatagtccg gaatcgtcac
     4441 acaaaaaacc aacacacaga tgtaatgaaa ataaagatat tttattgcgg ccatcgtgat
     4501 ggctagagtt tagccctccc acacataacc agagggcagc aattcacgaa tcccaactgc
     4561 cgtcggctgt ccatcactgt ccttcactat ggctttgatc ccaggatgca gatcgagaag
     4621 cacctgtcgg caccgtccgc aggggctcaa gatgcccctg ttctcatttc cgatcgcgac
     4681 gatacaagtc aggttgccag ctgccgcagc agcagcagtg cccagcacca cgagttctgc
     4741 acaaggtccc ccagtaaaat gatatacatt gacaccagtg aagatgcggc cgtcgctaga
     4801 gagagctgcg ctggcgacgc tgtagtcttc agagatgggg atgctgttga ttgtagccgt
     4861 tgctctttca atgagggtgg attcttcttg agacaaaggc ttggccatgg tggcgctagt
     4921 gtcagaagaa tcgagctttt tgcaaaagcc taggcctcca aaaaagcctc ctcactactt
     4981 ctggaatagc tcagaggccg aggcggcctc ggcctctgca taaataaaaa aaattagtca
     5041 gccatggggc ggagaatggg cggaactggg cggagttagg ggcgggatgg gcggagttag
     5101 gggcgggact atggttgctg actaattgag atgcatgctt tgcatacttc tgcctgctgg
     5161 ggagcctggg gactttccac acctggttgc tgactaattg agatgcatgc tttgcatact
     5221 tctgcctgct ggggagcctg gggactttcc acaccctaac tgacacacat tccacagctg
     5281 gttctttccg cctcagaagg tacctaacca agttcctctt tcagaggtta tttcaggcca
     5341 tggtgctgcg c
//
