LOCUS       Exported                6474 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 6474)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Apr 15, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
COMMENT     pRSG194 designed by Brock F. Binkowski

            

            PCR amplify pF-BiBiTsmC 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..6474
                     /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
     misc_feature    1180..1187
                     /label=SgfI Site
                     /note="Used in Flexi Vector Cloning"
     misc_feature    1189..3360
                     /label=PIK3R1
                     /note="PIK3R1"
     CDS             3370..3414
                     /codon_start=1
                     /label=15GS linker
                     /note="15GS linker"
                     /translation="GSSGGGGSGGGGSSG"
     CDS             3415..3447
                     /codon_start=1
                     /label=SmBiT
                     /note="SmBiT"
                     /translation="VTGYRLFEEIL"
     polyA_signal    3528..3749
                     /label=Sv40 late polyadenylation signal
     misc_feature    complement(3773..3806)
                     /label=Cre site
                     /note="Cre site"
     promoter        3815..3881
                     /label=EM7 bacterial promoter
     CDS             3895..4689
                     /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      4844..4880
                     /direction=RIGHT
                     /label=ColE1-derived plasmid replication origin
                     /note="The actual origin is the first base."
     polyA_signal    complement(5562..5609)
                     /label=Synthetic PolyAdenylation Signal
     misc_feature    complement(5633..6031)
                     /label=BlastR
                     /note="BlastR"
     misc_feature    complement(6056..6474)
                     /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 cgatcgccat gagtgctgag
     1201 gggtaccagt acagagcgct gtatgattat aaaaaggaaa gagaagaaga tattgacttg
     1261 cacttgggtg acatattgac tgtgaataaa gggtccttag tagctcttgg attcagtgat
     1321 ggacaggaag ccaggcctga agaaattggc tggttaaatg gctataatga aaccacaggg
     1381 gaaagggggg actttccggg aacttacgta gaatatattg gaaggaaaaa aatctcgcct
     1441 cccacaccaa agccccggcc acctcggcct cttcctgttg caccaggttc ttcgaaaact
     1501 gaagcagatg ttgaacaaca agctttgact ctcccggatc ttgcagagca gtttgcccct
     1561 cctgacattg ccccgcctct tcttatcaag ctcgtggaag ccattgaaaa gaaaggtctg
     1621 gaatgttcaa ctctatacag aacacagagc tccagcaacc tggcagaatt acgacagctt
     1681 cttgattgtg atacaccctc cgtggacttg gaaatgatcg atgtgcacgt tttggctgac
     1741 gctttcaaac gctatctcct ggacttacca aatcctgtca ttccagcagc cgtttacagt
     1801 gaaatgattt ctttagctcc agaagtacaa agctccgaag aatatattca gctattgaag
     1861 aagcttatta ggtcgcctag catacctcat cagtattggc ttacgcttca gtatttgtta
     1921 aaacatttct tcaagctctc tcaaacctcc agcaaaaatc tgttgaatgc aagagtactc
     1981 tctgaaattt tcagccctat gcttttcaga ttctcagcag ccagctctga taatactgaa
     2041 aacctcataa aagttataga aattttaatc tcaactgaat ggaatgaacg acagcctgca
     2101 ccagcactgc ctcctaaacc accaaaacct actactgtag ccaacaacgg tatgaataac
     2161 aatatgtcct tacaagatgc tgaatggtac tggggagata tctcgaggga agaagtgaat
     2221 gaaaaacttc gagatacagc agacgggacc tttttggtac gagatgcgtc tactaaaatg
     2281 catggtgatt atactcttac actaaggaaa gggggaaata acaaattaat caaaatattt
     2341 catcgagatg ggaaatatgg cttctctgac ccattaacct tcagttctgt ggttgaatta
     2401 ataaaccact accggaatga atctctagct cagtataatc ccaaattgga tgtgaaatta
     2461 ctttatccag tatccaaata ccaacaggat caagttgtca aagaagataa tattgaagct
     2521 gtagggaaaa aattacatga atataacact cagtttcaag aaaaaagtcg agaatatgat
     2581 agattatatg aagaatatac ccgcacatcc caggaaatcc aaatgaaaag gacagctatt
     2641 gaagcattta atgaaaccat aaaaatattt gaagaacagt gccagaccca agagcggtac
     2701 agcaaagaat acatagaaaa gttcaaacgt gaaggcaatg agaaagaaat acaaaggatt
     2761 atgcataatt atgataagtt gaagtctcga atcagtgaaa ttattgacag tagaagaaga
     2821 ttggaagaag acttgaagaa gcaggcagct gagtatcgag aaattgacaa acgtatgaac
     2881 agcattaaac cagaccttat ccagctgaga aagacgagag accaatactt gatgtggttg
     2941 actcaaaaag gtgttcggca aaagaagttg aacgagtggt tgggcaatga aaacactgaa
     3001 gaccaatatt cactggtgga agatgatgaa gatttgcccc atcatgatga gaagacatgg
     3061 aatgttggaa gcagcaaccg aaacaaagct gaaaacctgt tgcgagggaa gcgagatggc
     3121 acttttcttg tccgggagag cagtaaacag ggctgctatg cctgctctgt agtggtggac
     3181 ggcgaagtaa agcattgtgt cataaacaaa acagcaactg gctatggctt tgccgagccc
     3241 tataacttgt acagctctct gaaagaactg gtgctacatt accaacacac ctcccttgtg
     3301 cagcacaacg actccctcaa tgtcacacta gcctacccag tatatgcaca gcagaggcga
     3361 gtttctcaag gcagttcagg tggtggcggg agcggaggtg gaggctcgag cggtgtgacc
     3421 ggctaccggc tgttcgagga gattctgtaa ggccgcgact ctagagtcga cctgcaggca
     3481 tgcaagctga tttggctgct gccaccgctg agcaaggccg cttcgagcag acatgataag
     3541 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
     3601 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
     3661 caacaacaat tgcattcatt ttatgtttca ggttcagggg gagatgtggg aggttttttt
     3721 aagcaagtaa aacctctaca aatgtggtaa aatcttaatt aagaattcta atataacttc
     3781 gtatagcata cattatacga agttatggat ccactgttga caattaatca tcggcatagt
     3841 atatcggcat agtataatac gacaaggtga ggaactaaac ccaggaggca gatcatgatt
     3901 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     3961 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     4021 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     4081 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     4141 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     4201 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     4261 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     4321 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     4381 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     4441 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     4501 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     4561 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     4621 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     4681 ttcttctgat tcgaaatgac cgaccaagcg acgcccaacc ggtatcagct cactcaaagg
     4741 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     4801 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     4861 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     4921 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     4981 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     5041 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     5101 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     5161 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     5221 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     5281 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     5341 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     5401 agcagcagat tacgcgcaga aaaaaaggat ttcaagaaga tcctttgatc ttttctacgg
     5461 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     5521 aaaggatctt cacctagatc cttttatagt ccggaatcgt cacacaaaaa accaacacac
     5581 agatgtaatg aaaataaaga tattttattg cggccatcgt gatggctaga gtttagccct
     5641 cccacacata accagagggc agcaattcac gaatcccaac tgccgtcggc tgtccatcac
     5701 tgtccttcac tatggctttg atcccaggat gcagatcgag aagcacctgt cggcaccgtc
     5761 cgcaggggct caagatgccc ctgttctcat ttccgatcgc gacgatacaa gtcaggttgc
     5821 cagctgccgc agcagcagca gtgcccagca ccacgagttc tgcacaaggt cccccagtaa
     5881 aatgatatac attgacacca gtgaagatgc ggccgtcgct agagagagct gcgctggcga
     5941 cgctgtagtc ttcagagatg gggatgctgt tgattgtagc cgttgctctt tcaatgaggg
     6001 tggattcttc ttgagacaaa ggcttggcca tggtggcgct agtgtcagaa gaatcgagct
     6061 ttttgcaaaa gcctaggcct ccaaaaaagc ctcctcacta cttctggaat agctcagagg
     6121 ccgaggcggc ctcggcctct gcataaataa aaaaaattag tcagccatgg ggcggagaat
     6181 gggcggaact gggcggagtt aggggcggga tgggcggagt taggggcggg actatggttg
     6241 ctgactaatt gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc
     6301 cacacctggt tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc
     6361 ctggggactt tccacaccct aactgacaca cattccacag ctggttcttt ccgcctcaga
     6421 aggtacctaa ccaagttcct ctttcagagg ttatttcagg ccatggtgct gcgc
//
