LOCUS       Exported                6383 bp ds-DNA     circular SYN 30-MAY-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6383)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Friday, May 30, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
COMMENT     pRSG195 designed by Brock F. Binkowski

            

            PCR amplify pF-BiBiTsmN w/ BFBRSG15/BFBRSG14 (PCR #1; 3.9 kbp);

            PCR amplify pF-4B w/ BFBRSG13/BFBRSG16 (PCR #2; 871 bp); 

            2X component Gibson Assembly;

            electroporation of BR610.
FEATURES             Location/Qualifiers
     source          1..6383
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    complement(22..209)
                     /label=SV40\polyA\signal
     misc_feature    complement(213..275)
                     /label=MCS 2
     promoter        276..866
                     /label=CMV promoter
                     /note="Has EcoICRI site removed. Named changed from ""CMV 
                     Promoter"" to ""CMV immediate-early enhancer/promoter"" 
                     3/21/2011 and J Hartnet's request to fit Sci Com's naming."
     intron          981..1113
                     /label=chimeric intron
                     /note="Note that this is the actual sequence spliced out. 
                     The 5' donor site includes a 5' GA and the 3' acceptor site
                     adds a 3'G."
     promoter        1157..1176
                     /label=T7 promoter
     CDS             1189..1224
                     /codon_start=1
                     /label=SmBiT
                     /note="SmBiT"
                     /translation="MVTGYRLFEEIL"
     CDS             1225..1269
                     /codon_start=1
                     /label=15GS linker
                     /note="15GS linker"
                     /translation="GSSGGGGSGGGGSSG"
     misc_feature    1270..1277
                     /label=SgfI Site
                     /note="Used in Flexi Vector Cloning"
     CDS             1279..3300
                     /label=PPP1R15A
     misc_feature    3301..3308
                     /label=PmeI site
                     /note="Used in Flexi Vector Cloning"
     polyA_signal    3437..3658
                     /label=Sv40 late polyadenylation signal
     promoter        3724..3790
                     /label=EM7 bacterial promoter
     CDS             3804..4598
                     /label=KanR
                     /note="Name changed frm ""Neomycin phosphotransferase"" to 
                     ""Neomycin phosphotransferase coding region"" on 3/21/2011 
                     at Jim Hartnet's request to fit Sci Com's naming."
     rep_origin      4753..4789
                     /direction=RIGHT
                     /label=ColE1-derived plasmid replication origin
                     /note="The actual origin is the first base."
     polyA_signal    complement(5471..5518)
                     /label=Synthetic PolyAdenylation Signal
     misc_feature    complement(5542..5940)
                     /label=BlastR
                     /note="BlastR"
     misc_feature    complement(5965..6383)
                     /label=SV40 promoter
                     /note="SV40 promoter"
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 ggccccaggc caagcacccc atcaggctac cccgtggagg
     1321 gatgcccacc ctttcttcct cctgtcccca gtgatgggcc tcctcagccg cgcctggagc
     1381 cgcctgaggg gcctgggacc tctagagccc tggctggtgg aagcagtaaa aggagcagct
     1441 ctggtagaag ctggcctgga gggagaagct aggactcctc tggcaatccc ccatacccct
     1501 tggggcagac gccctgaaga ggaggctgaa gacagtggag gccctggaga ggacagagaa
     1561 acactggggc tgaaaaccag cagttccctt cctgaagcct ggggactttt ggatgatgat
     1621 gatggcatgt atggtgagcg agaggcaacc agtgtcccta gagggcaggg aagtcaattt
     1681 gcagatggcc agcgtgctcc cctgtctccc agccttctga taaggacact gcaaggttct
     1741 gataagaacc caggggagga gaaagccgag gaagagggag ttgctgaaga ggagggagtt
     1801 aacaagttct cttatccacc atcacaccgg gagtgttgtc cagccgtgga ggaggaggac
     1861 gatgaagaag ctgtaaagaa agaagctcac agaacctcta cttctgcctt gtctccagga
     1921 tccaagccca gcacttgggt gtcttgccca ggggaggaag agaatcaagc cacggaggat
     1981 aaaagaacag aaagaagtaa aggagccagg aagacctccg tgtccccccg atcttcaggc
     2041 tccgacccca ggtcctggga gtatcgttca ggagaggcgt ccgaggagaa ggaggaaaag
     2101 gcacacaaag aaactgggaa aggagaagct gccccagggc cgcaatcctc agccccagcc
     2161 cagaggcccc agctcaagtc ctggtggtgc caacccagtg atgaagagga gggtgaggtc
     2221 aaggctttgg gggcagctga gaaggatgga gaagctgagt gtcctccctg catcccccca
     2281 ccaagtgcct tcctgaaggc ctgggtgtat tggccaggag aggacacaga ggaagaggaa
     2341 gatgaggaag aagatgagga cagtgactct ggatcagatg aggaagaggg agaagctgag
     2401 gcttcctctt ccactcctgc tacaggtgtc ttcttgaagt cctgggtcta tcagccagga
     2461 gaggacacag aggaggagga agatgaggac agtgatacag gatcagccga ggatgaaaga
     2521 gaagctgaga cttctgcttc cacaccccct gcaagtgctt tcttgaaggc ctgggtgtat
     2581 cggccaggag aggacacgga ggaggaggaa gatgaggatg tggatagtga ggataaggaa
     2641 gatgattcag aagcagcctt gggagaagct gagtcagacc cacatccctc ccacccggac
     2701 cagagggccc acttcagggg ctggggatat cgacctggaa aagagacaga ggaagaggaa
     2761 gctgctgagg actggggaga agctgagccc tgccccttcc gagtggccat ctatgtacct
     2821 ggagagaagc caccgcctcc ctgggctcct cctaggctgc ccctccgact gcaaaggcgg
     2881 ctcaagcgcc cagaaacccc tactcatgat ccggaccctg agactcccct aaaggccaga
     2941 aaggtgcgct tctccgagaa ggtcactgtc catttcctgg ctgtctgggc agggccggcc
     3001 caggccgccc gccagggccc ctgggagcag cttgctcggg atcgcagccg cttcgcacgc
     3061 cgcatcaccc aggcccagga ggagctgagc ccctgcctca cccctgctgc ccgggccaga
     3121 gcctgggcac gcctcaggaa cccaccttta gcccccatcc ctgccctcac ccagaccttg
     3181 ccttcctcct ctgtcccttc gtccccagtc cagaccacgc ccttgagcca agctgtggcc
     3241 acaccttccc gctcgtctgc tgctgcagcg gctgccctgg acctcagtgg gaggcgtggc
     3301 gtttaaacga attcgggctc ggtacccggg gatcctctag agtcgacctg caggcatgca
     3361 agctgatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga
     3421 gcaaggccgc ttcgagcaga catgataaga tacattgatg agtttggaca aaccacaact
     3481 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
     3541 accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag
     3601 gttcaggggg agatgtggga ggttttttta agcaagtaaa acctctacaa atgtggtaaa
     3661 atcttaatta agaattctaa tataacttcg tatagcatac attatacgaa gttatggatc
     3721 cactgttgac aattaatcat cggcatagta tatcggcata gtataatacg acaaggtgag
     3781 gaactaaacc caggaggcag atcatgattg aacaagatgg attgcacgca ggttctccgg
     3841 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     3901 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     3961 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
     4021 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     4081 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     4141 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     4201 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     4261 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     4321 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     4381 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     4441 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     4501 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     4561 gcatcgcctt ctatcgcctt cttgacgagt tcttctgatt cgaaatgacc gaccaagcga
     4621 cgcccaaccg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     4681 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     4741 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     4801 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     4861 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     4921 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     4981 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     5041 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     5101 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     5161 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
     5221 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     5281 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatt
     5341 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     5401 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttatagtc
     5461 cggaatcgtc acacaaaaaa ccaacacaca gatgtaatga aaataaagat attttattgc
     5521 ggccatcgtg atggctagag tttagccctc ccacacataa ccagagggca gcaattcacg
     5581 aatcccaact gccgtcggct gtccatcact gtccttcact atggctttga tcccaggatg
     5641 cagatcgaga agcacctgtc ggcaccgtcc gcaggggctc aagatgcccc tgttctcatt
     5701 tccgatcgcg acgatacaag tcaggttgcc agctgccgca gcagcagcag tgcccagcac
     5761 cacgagttct gcacaaggtc ccccagtaaa atgatataca ttgacaccag tgaagatgcg
     5821 gccgtcgcta gagagagctg cgctggcgac gctgtagtct tcagagatgg ggatgctgtt
     5881 gattgtagcc gttgctcttt caatgagggt ggattcttct tgagacaaag gcttggccat
     5941 ggtggcgcta gtgtcagaag aatcgagctt tttgcaaaag cctaggcctc caaaaaagcc
     6001 tcctcactac ttctggaata gctcagaggc cgaggcggcc tcggcctctg cataaataaa
     6061 aaaaattagt cagccatggg gcggagaatg ggcggaactg ggcggagtta ggggcgggat
     6121 gggcggagtt aggggcggga ctatggttgc tgactaattg agatgcatgc tttgcatact
     6181 tctgcctgct ggggagcctg gggactttcc acacctggtt gctgactaat tgagatgcat
     6241 gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacccta actgacacac
     6301 attccacagc tggttctttc cgcctcagaa ggtacctaac caagttcctc tttcagaggt
     6361 tatttcaggc catggtgctg cgc
//
