LOCUS       Exported                8878 bp ds-DNA     circular SYN 20-SEP-2024
DEFINITION  sgPPP1R2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8878)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Sep 20, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8878
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     LTR             651..831
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     1341..1357
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1360..1948
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             1466..1477
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    complement(2004..2037)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     CDS             complement(2057..2755)
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="SELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKVV
                     EGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTAT
                     QDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALKL
                     VGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYCD
                     LPSKLGHKLN"
     CDS             complement(2768..2821)
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             complement(2831..3427)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     intron          3448..4386
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     promoter        complement(4387..4598)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     protein_bind    complement(4868..4901)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    5154..5271
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(5569..5613)
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    5798..6031
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    6528..6653
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             6700..6880
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(6898..7096)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     enhancer        7098..7477
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7743..7847
                     /gene="bla"
                     /label=AmpR promoter
     CDS             7848..8708
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgc tgctagagat
      661 tttccacact gactaaaagg gtctgaggga tctctagtta ccagagtcac acaacagacg
      721 ggcacacact acttgaagca ctcaaggcaa gctttattga ggcttaagca gtgggttccc
      781 tagttagcca gagagctccc aggctcagat ctggtctaac cagagagacc cagtacagtc
      841 cggatgcagc tctcgggcca tgtgatgaaa tgctaggcgg ctgtcaaacc tccactctaa
      901 tacttctctc tccgggtcat ccatcccatg caggctcaca gggtgtaaca agcgggtgtt
      961 ctctccttca ttggcttctt ctaccttctc ttgctcaact ggtactagct tgtagcacca
     1021 tccaaaggtc agtggatatc tgatccctgg ccctggtgtg tagttctgcc aatcagggaa
     1081 gtagccttgt gtgtggtaga tccacagatc aaggatatct tgtcttcgtt gggagtgaat
     1141 tagcccttcc agtcccccct tttcttttaa aaagtggcta agatctacag ctgccttgta
     1201 agtcattggt cttaaaggta cctgaggtgt gactggaaaa cccacctcct cctcctcttg
     1261 tgcttctagc caggcacaat cagcattggt agctgctgta ttgctacttg tgattgctcc
     1321 atgtttttct aggtctcgat cgaggtcgac ggtatcgatg cggggaggcg gcccaaaggg
     1381 agatccgact cgtctgaggg cgaaggcgaa gacgcggaag aggccgcaga gccggcagca
     1441 ggccgcggga aggaaggtcc gctggattga gggccgaagg gacgtagcag aaggacgtcc
     1501 cgcgcagaat ccaggtggca acacaggcga gcagccaagg aaaggacgat gatttccccg
     1561 acaacaccac ggaattgtca gtgcccaaca gccgagcccc tgtccagcag cgggcaaggc
     1621 aggcggcgat gagttccgcc gtggcaatag ggagggggaa agcgaaagtc ccggaaagga
     1681 gctgacaggt ggtggcaatg ccccaaccag tgggggttgc gtcagcaaac acagtgcaca
     1741 ccacgccacg ttgcctgaca acgggccaca actcctcata aagagacagc aaccaggatt
     1801 tatacaagga ggagaaaatg aaagccatac gggaagcaat agcatgatac aaaggcatta
     1861 aagcagcgta tccacatagc gtaaaaggag caacatagtt aagaatacca gtcaatcttt
     1921 cacaaatttt gtaatccaga ggttgattat cgataagctt gatatcgaat tgtacctagt
     1981 ggaaccggaa cccttaaaca tgtataactt cgtataatgt atgctatacg aagttattag
     2041 gtccctcgac gaattcttaa ttaagcttgt gccccagttt gctagggagg tcgcagtatc
     2101 tggccactgc cacctcgtgc tgctcgacgt aggtctcgtt gttggcctcc ttgattcttt
     2161 ccagtctgta gtccacatag tagacgccag gcatcttgag gttcttagcg ggtttcttgg
     2221 atctatatgt ggtcttgatg tttgcgatca gatggctccc gcccacgagc ttcagggcca
     2281 tgtcgtttct gccttccagg ccgccgtcag cggggtacag cgtctcggtg aaggcctccc
     2341 agccgagtgt tttcttctgc atcacagggc cgttggatgt gaagttcacc cctctgatct
     2401 tgacgttgta gatgaggcag ccgtcctgga ggctggtgtc ctgggtagcg gtcagcacgc
     2461 ccccgtcttc gtatgtggtg actctctccc atgtgaagcc ctcagggaag gactgcttga
     2521 agaagtcggg gatgccctgg gtgtggttga tgaaggtctt gctgccgtag aggaagctag
     2581 tagccaggat gtcgaaggcg aaggggagag ggccgccctc gaccaccttg attctcatgg
     2641 tctgggtgcc ctcgtagggc ttgccttcgc cctcggatgt gcacttgaag tgatggttgt
     2701 ccacggtgcc ctccatgtac agcttcatgt gcatgttctc cttaatcagc tcgctggtgg
     2761 cgaccggtgg gccaggattc tcctcgacgt caccgcatgt tagcagactt cctctgccct
     2821 ctcccgatcc ggcaccgggc ttgcgggtca tgcaccaggt gcgcggtcct tcgggcacct
     2881 cgacgtcggc ggtgacggtg aagccgagcc gctcgtagaa ggggaggttg cggggcgcgg
     2941 aggtctccag gaaggcgggc accccggcgc gctcggccgc ctccactccg gggagcacga
     3001 cggcgctgcc cagacccttg ccctggtggt cgggcgagac gccgacggtg gccaggaacc
     3061 acgcgggctc cttgggccgg tgcggcgcca ggaggccttc catctgttgc tgcgcggcca
     3121 gccgggaacc gctcaactcg gccatgcgcg ggccgatctc ggcgaacacc gcccccgctt
     3181 cgacgctctc cggcgtggtc cagaccgcca ccgcggcgcc gtcgtccgcg acccacacct
     3241 tgccgatgtc gagcccgacg cgcgtgagga agagttcttg cagctcggtg acccgctcga
     3301 tgtggcggtc cgggtcgacg gtgtggcgcg tggcggggta gtcggcgaac gcggcggcga
     3361 gggtgcgtac ggcccggggg acgtcgtcgc gggtggcgag gcgcaccgtg ggcttgtact
     3421 cggtcatggt gggctagctc acgacacctg aaatggaaga aaaaaacttt gaaccactgt
     3481 ctgaggcttg agaatgaacc aagatccaaa ctcaaaaagg gcaaattcca aggagaatta
     3541 catcaagtgc caagctggcc taacttcagt ctccacccac tcagtgtggg gaaactccat
     3601 cgcataaaac ccctcccccc aacctaaaga cgacgtactc caaaagcgcg agaactaatc
     3661 gaggtgcctg gacggcgccc ggtactccgt ggagtcacat gaagcgacgg ctgaggacgg
     3721 aaaggccctt ttcctttgtg tgggtgactc acccgcccgc tctcccgagc gccgcgtcct
     3781 ccattttgag ctccctgcag cagggccggg aagcggccat ctttccgctc acgcaactgg
     3841 tgccgaccgg gccagccttg ccgcccaggg cggggcgata cacggcggcg cgaggccagg
     3901 caccagagca ggccggccag cttgagacta cccccgtccg attctcggtg gccgcgctcg
     3961 caggccccgc ctcgccgaac atgtgcgctg ggacgcacgg gccccgtcgc cgcccgcggc
     4021 cccaaaaacc gaaataccag tgtgcagatc ttggcccgca tttacaagac tatcttgcca
     4081 gaaaaaaagc gtcgcagcag gtcatcaaaa attttaaatg gctagagact tatcgaaagc
     4141 agcgagacag gcgcgaaggt gccaccagat tcgcacgcgg cggccccagc gcccaggcca
     4201 ggcctcaact caagcacgag gcgaaggggc tccttaagcg caaggcctcg aactctccca
     4261 cccacttcca acccgaagct cgggatcaag aatcacgtac tgcagccagg tggaagtaat
     4321 tcaaggcacg caagggccat aacccgtaaa gaggccaggc ccgcgggaac cacacacggc
     4381 acttacctgt gttctggcgg caaacccgtt gcgaaaaaga acgttcacgg cgactactgc
     4441 acttatatac ggttctcccc caccctcggg aaaaaggcgg agccagtaca cgacatcact
     4501 ttcccagttt accccgcgcc accttctcta ggcaccggtt caattgccga cccctccccc
     4561 caacttctcg gggactgtgg gcgatgtgcg ctctgcccac tgacgggcac atgcatggcg
     4621 gtaatacggt tatccacgcg gccgcctaat ggatcctagt actcgagaaa aaaagcaccg
     4681 actcggtgcc actttttcaa gttgataacg gactagcctt atttaaactt gctatgctgt
     4741 ttccagcata gctcttaaac ctgtgatatc atacagagga ccaacaaggt ggttctccaa
     4801 gggatactta tagtctcaaa acacacaatt actttacagt tagggtgagt ttccttttgt
     4861 gctgtttata acttcgtata atgtatgcta tacgaagtta tagaaatcca agcctatcat
     4921 gtaaaatgta gctagtatta aaaagaacag attatctgtc ttttatcgca cattaagcct
     4981 ctatagttac taggaaatat tatatgcaaa ttaaccgggg caggggagta gccgagcttc
     5041 tcccacaagt ccgtgcgagg gggccggcgc gggcctagag atggcgcgtc ggatctctag
     5101 agcgggcccg gtactaacca aactggatct ctgctgtccc tgtaataaac ccgaaaattt
     5161 tgaatttttg taatttgttt ttgtaattct ttagtttgta tgtctgttgc tattatgtct
     5221 actattcttt cccctgcact gtacccccca atcccccctt ttcttttaaa attgtggatg
     5281 aatactgcca tttgtctgca gaattggcgc acgcagtgcc gatccgttca ctaatcgaat
     5341 ggatctgtct ctgtctctct ctccaccttc ttcttctatt ccttcgggcc tgtcgggtcc
     5401 cctcggggtt gggaggtggg tctgaaacga taatggtgaa tatccctgcc taactctatt
     5461 cactatagaa agtacagcaa aaactattct taaacctacc aagcctccta ctatcattat
     5521 gaataatttt atataccaca gccaatttgt tatgttaaac caattccaca aacttgccca
     5581 tttatctaat tccaataatt cttgttcatt cttttcttgc tggttttgcg attcttcaat
     5641 taaggagtgt attaagcttg tgtaattgtt aatttctctg tcccactcca tccaggtcgt
     5701 gtgattccaa atctgttcca gagatttatt actccaacta gcattccaag gcacagcagt
     5761 ggtgcaaatg agttttccag agcaacccca aatccccagg agctgttgat cctttaggta
     5821 tctttccaca gccaggattc ttgcctggag ctgcttgatg ccccagactg tgagttgcaa
     5881 cagatgctgt tgcgcctcaa tagccctcag caaattgttc tgctgctgca ctataccaga
     5941 caataattgt ctggcctgta ccgtcagcgt cattgacgct gcgcccatag tgcttcctgc
     6001 tgctcccaag aacccaagga acaaagctcc tattcccact gctctttttt ctctctgcac
     6061 cactcttctc tttgccttgg tgggtgctac tcctaatggt tcaattttta ctactttata
     6121 tttatataat tcacttctcc aattgtccct catatctcct cctccaggtc tgaagatcag
     6181 cggccggccg cttgctgtgc ggtggtctta cttttgtttt gctcttcctc tatcttgtct
     6241 aaagcttcct tggtgtcttt tatctctatc ctttgatgca cacaatagag ggttgctact
     6301 gtattatata atgatctaag ttcttctgat cctgtctgaa gggatggttg tagctgtccc
     6361 agtatttgtc tacagccttc tgatgtttct aacaggccag gattaactgc gaatcgttct
     6421 agctccctgc ttgcccatac tatatgtttt aatttatatt ttttctttcc ccctggcctt
     6481 aaccgaattt tttcccatcg cgatctaatt ctcccccgct taatactgac gctctcgcac
     6541 ccatctctct ccttctagcc tccgctagtc aaaatttttg gcgtactcac cagtcgccgc
     6601 ccctcgcctc ttgccgtgcg cgcttcagca agccgagtcc tgcgtcgaga gagctcctct
     6661 ggtttccctt tcgctttcaa gtccctgttc gggcgccact gctagagatt ttccacactg
     6721 actaaaaggg tctgagggat ctctagttac cagagtcaca caacagacgg gcacacacta
     6781 cttgaagcac tcaaggcaag ctttattgag gcttaagcag tgggttccct agttagccag
     6841 agagctccca ggctcagatc tggtctaacc agagagaccc agtacaggca aaacgcgctg
     6901 cttatataga cctcccaccg tacacgccta ccgcccattt gcgtcaatgg ggcggagttg
     6961 ttacgacatt ttggaaagtc ccgttgattt tggtgccaaa acaaactccc attgacgtca
     7021 atggggtgga gacttggaaa tccccgtgag tcaaaccgct atccacgccc attgatgtac
     7081 tgccaaaacc gcatcaccat ggtaatagcg atgactaata cgtagatgta ctgccaagta
     7141 ggaaagtccc ataaggtcat gtactgggca taatgccagg cgggccattt accgtcattg
     7201 acgtcaatag ggggcgtact tggcatatga tacacttgat gtactgccaa gtgggcagtt
     7261 taccgtaaat actccaccca ttgacgtcaa tggaaagtcc ctattggcgt tactatggga
     7321 acatacgtca ttattgacgt caatgggcgg gggtcgttgg gcggtcagcc aggcgggcca
     7381 tttaccgtaa gttatgtaac gcggaactcc atatatgggc tatgaactaa tgaccccgta
     7441 attgattact attaataact agtcaataat caatgtcaac gcgtatatct ggcccgtaca
     7501 tcgcgaagca gcgcaaaacg cctaacccta agcagattct tcatgcaatt gtcggtcaag
     7561 ccttgccttg ttgtagctta aattttgctc gcgcactact cagcgacctc caacacacaa
     7621 gcagggagca gatactggct taactatgcg gcatcagagc agattgtact gagagtgcac
     7681 cataggggat cgggagatct cccgatccgt cgacgtcagg tggcactttt cggggaaatg
     7741 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     7801 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     7861 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     7921 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     7981 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     8041 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     8101 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     8161 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     8221 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     8281 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     8341 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     8401 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     8461 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     8521 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     8581 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     8641 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     8701 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     8761 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     8821 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttc
//
