LOCUS       Exported                5613 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 5613)
  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..5613
                     /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..2499
                     /label=VPS4A
                     /note="VPS4A"
     CDS             2509..2553
                     /codon_start=1
                     /label=15GS linker
                     /note="15GS linker"
                     /translation="GSSGGGGSGGGGSSG"
     CDS             2554..2586
                     /codon_start=1
                     /label=SmBiT
                     /note="SmBiT"
                     /translation="VTGYRLFEEIL"
     polyA_signal    2667..2888
                     /label=Sv40 late polyadenylation signal
     promoter        2954..3020
                     /label=EM7 bacterial promoter
     CDS             3034..3828
                     /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      3983..4019
                     /direction=RIGHT
                     /label=ColE1-derived plasmid replication origin
                     /note="The actual origin is the first base."
     polyA_signal    complement(4701..4748)
                     /label=Synthetic PolyAdenylation Signal
     misc_feature    complement(4772..5170)
                     /label=BlastR
                     /note="BlastR"
     misc_feature    complement(5195..5613)
                     /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 gacaacgtca
     1201 accctccaga aagccattga tctggtgacg aaagccacag aggaggacaa agccaagaac
     1261 tacgaggagg cgctgcggct gtaccagcat gcggtggagt acttcctcca cgctatcaag
     1321 tatgaggccc acagcgacaa ggccaaggag agcattcgag ccaagtgcgt gcagtaccta
     1381 gaccgggccg agaagctgaa ggattattta cgaagcaaag agaaacacgg caagaagcca
     1441 gtcaaagaga accagagtga gggcaagggc agtgacagtg acagtgaagg ggataatccg
     1501 gagaaaaaga aactgcaaga acagctgatg ggtgccgtcg tgatggagaa gcccaacata
     1561 cggtggaacg acgtggccgg gctggagggg gccaaggagg ccctcaaaga agctgtcatt
     1621 ttgccaatca aattcccaca cttgttcaca ggcaagcgca ccccctggcg ggggattctg
     1681 ctgttcggac cccctggcac agggaaatcc tacctggcca aagccgtggc aacagaggcc
     1741 aacaactcca ccttcttctc tgtgtcctcc tcagatctga tgtccaagtg gctgggggag
     1801 agtgaaaagc tggtcaagaa cctgtttgag ctggccaggc agcacaagcc ctccatcatc
     1861 ttcatcgatg aggtggattc cctctgcggg tcccgaaatg aaaatgagag tgaggccgcc
     1921 cggaggatca aaacggagtt cttggtccag atgcaggggg tggggaataa caatgatggg
     1981 actctggttc ttggagccac aaacatccca tgggtgttgg attcggccat caggaggagg
     2041 tttgaaaaac gaatttatat ccccttgccg gaggaagctg cccgcgccca gatgttccgg
     2101 ttgcatctcg ggagcactcc ccacaacctc acggatgcaa acatccacga gctggcccgg
     2161 aagacggaag gctactcggg cgcggacatc agcatcatcg tgcgggactc tctcatgcag
     2221 cccgtgagga aggtgcagtc ggccacacac ttcaaaaagg tctgtggccc ctctcgcacc
     2281 aaccccagca tgatgattga tgacctcctg actccatgct caccagggga cccaggagcc
     2341 atggagatga cttggatgga tgtccctggg gacaaactct tagagcctgt ggtttgcatg
     2401 tcggacatgc tgcggtctct ggccaccacc cggcccacgg tgaatgcaga cgacctcctg
     2461 aaagtgaaga aattctcaga ggactttggg caagagagtg tttctcaagg cagttcaggt
     2521 ggtggcggga gcggaggtgg aggctcgagc ggtgtgaccg gctaccggct gttcgaggag
     2581 attctgtaag gccgcgactc tagagtcgac ctgcaggcat gcaagctgat ttggctgctg
     2641 ccaccgctga gcaaggccgc ttcgagcaga catgataaga tacattgatg agtttggaca
     2701 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     2761 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
     2821 tatgtttcag gttcaggggg agatgtggga ggttttttta agcaagtaaa acctctacaa
     2881 atgtggtaaa atcttaatta agaattctaa tataacttcg tatagcatac attatacgaa
     2941 gttatggatc cactgttgac aattaatcat cggcatagta tatcggcata gtataatacg
     3001 acaaggtgag gaactaaacc caggaggcag atcatgattg aacaagatgg attgcacgca
     3061 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     3121 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     3181 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
     3241 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     3301 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     3361 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     3421 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     3481 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     3541 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc
     3601 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     3661 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     3721 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     3781 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgatt cgaaatgacc
     3841 gaccaagcga cgcccaaccg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     3901 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     3961 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     4021 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     4081 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     4141 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     4201 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     4261 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     4321 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     4381 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     4441 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     4501 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     4561 aaaaaggatt tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     4621 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     4681 ttttatagtc cggaatcgtc acacaaaaaa ccaacacaca gatgtaatga aaataaagat
     4741 attttattgc ggccatcgtg atggctagag tttagccctc ccacacataa ccagagggca
     4801 gcaattcacg aatcccaact gccgtcggct gtccatcact gtccttcact atggctttga
     4861 tcccaggatg cagatcgaga agcacctgtc ggcaccgtcc gcaggggctc aagatgcccc
     4921 tgttctcatt tccgatcgcg acgatacaag tcaggttgcc agctgccgca gcagcagcag
     4981 tgcccagcac cacgagttct gcacaaggtc ccccagtaaa atgatataca ttgacaccag
     5041 tgaagatgcg gccgtcgcta gagagagctg cgctggcgac gctgtagtct tcagagatgg
     5101 ggatgctgtt gattgtagcc gttgctcttt caatgagggt ggattcttct tgagacaaag
     5161 gcttggccat ggtggcgcta gtgtcagaag aatcgagctt tttgcaaaag cctaggcctc
     5221 caaaaaagcc tcctcactac ttctggaata gctcagaggc cgaggcggcc tcggcctctg
     5281 cataaataaa aaaaattagt cagccatggg gcggagaatg ggcggaactg ggcggagtta
     5341 ggggcgggat gggcggagtt aggggcggga ctatggttgc tgactaattg agatgcatgc
     5401 tttgcatact tctgcctgct ggggagcctg gggactttcc acacctggtt gctgactaat
     5461 tgagatgcat gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacccta
     5521 actgacacac attccacagc tggttctttc cgcctcagaa ggtacctaac caagttcctc
     5581 tttcagaggt tatttcaggc catggtgctg cgc
//
