LOCUS       Exported                5351 bp ds-DNA     circular SYN 15-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5351)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Apr 15, 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..5351
                     /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"
     misc_feature    1279..2268
                     /label=PPP1CA
                     /note="PPP1CA"
     misc_feature    2269..2276
                     /label=PmeI site
                     /note="Used in Flexi Vector Cloning"
     polyA_signal    2405..2626
                     /label=Sv40 late polyadenylation signal
     promoter        2692..2758
                     /label=EM7 bacterial promoter
     CDS             2772..3566
                     /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      3721..3757
                     /direction=RIGHT
                     /label=ColE1-derived plasmid replication origin
                     /note="The actual origin is the first base."
     polyA_signal    complement(4439..4486)
                     /label=Synthetic PolyAdenylation Signal
     misc_feature    complement(4510..4908)
                     /label=BlastR
                     /note="BlastR"
     misc_feature    complement(4933..5351)
                     /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 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
//
