LOCUS       Exported                4923 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 4923)
  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..4923
                     /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..1809
                     /label=CD3E
                     /note="CD3E"
     CDS             1819..1863
                     /codon_start=1
                     /label=15GS linker
                     /note="15GS linker"
                     /translation="GSSGGGGSGGGGSSG"
     CDS             1864..1896
                     /codon_start=1
                     /label=SmBiT
                     /note="SmBiT"
                     /translation="VTGYRLFEEIL"
     polyA_signal    1977..2198
                     /label=Sv40 late polyadenylation signal
     misc_feature    complement(2222..2255)
                     /label=Cre site
                     /note="Cre site"
     promoter        2264..2330
                     /label=EM7 bacterial promoter
     CDS             2344..3138
                     /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      3293..3329
                     /direction=RIGHT
                     /label=ColE1-derived plasmid replication origin
                     /note="The actual origin is the first base."
     polyA_signal    complement(4011..4058)
                     /label=Synthetic PolyAdenylation Signal
     misc_feature    complement(4082..4480)
                     /label=BlastR
                     /note="BlastR"
     misc_feature    complement(4505..4923)
                     /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 gcagtcgggc
     1201 actcactgga gagttctggg cctctgcctc ttatcagttg gcgtttgggg gcaagatggt
     1261 aatgaagaaa tgggtggtat tacacagaca ccatataaag tctccatctc tggaaccaca
     1321 gtaatattga catgccctca gtatcctgga tctgaaatac tatggcaaca caatgataaa
     1381 aacataggcg gtgatgagga tgataaaaac ataggcagtg atgaggatca cctgtcactg
     1441 aaggaatttt cagaattgga gcaaagtggt tattatgtct gctaccccag aggaagcaaa
     1501 ccagaagatg cgaactttta tctctacctg agggcaagag tgtgtgagaa ctgcatggag
     1561 atggatgtga tgtcggtggc cacaattgtc atagtggaca tctgcatcac tgggggcttg
     1621 ctgctgctgg tttactactg gagcaagaat agaaaggcca aggccaagcc tgtgacacga
     1681 ggagcgggtg ctggcggcag gcaaagggga caaaacaagg agaggccacc acctgttccc
     1741 aacccagact atgagcccat ccggaaaggc cagcgggacc tgtattctgg cctgaatcag
     1801 agacgcatcg tttctcaagg cagttcaggt ggtggcggga gcggaggtgg aggctcgagc
     1861 ggtgtgaccg gctaccggct gttcgaggag attctgtaag gccgcgactc tagagtcgac
     1921 ctgcaggcat gcaagctgat ttggctgctg ccaccgctga gcaaggccgc ttcgagcaga
     1981 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     2041 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     2101 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg agatgtggga
     2161 ggttttttta agcaagtaaa acctctacaa atgtggtaaa atcttaatta agaattctaa
     2221 tataacttcg tatagcatac attatacgaa gttatggatc cactgttgac aattaatcat
     2281 cggcatagta tatcggcata gtataatacg acaaggtgag gaactaaacc caggaggcag
     2341 atcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     2401 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     2461 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     2521 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     2581 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     2641 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     2701 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     2761 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     2821 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     2881 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     2941 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     3001 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     3061 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     3121 cttgacgagt tcttctgatt cgaaatgacc gaccaagcga cgcccaaccg gtatcagctc
     3181 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     3241 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     3301 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     3361 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     3421 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     3481 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     3541 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     3601 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     3661 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     3721 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     3781 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     3841 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatt tcaagaagat cctttgatct
     3901 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     3961 gattatcaaa aaggatcttc acctagatcc ttttatagtc cggaatcgtc acacaaaaaa
     4021 ccaacacaca gatgtaatga aaataaagat attttattgc ggccatcgtg atggctagag
     4081 tttagccctc ccacacataa ccagagggca gcaattcacg aatcccaact gccgtcggct
     4141 gtccatcact gtccttcact atggctttga tcccaggatg cagatcgaga agcacctgtc
     4201 ggcaccgtcc gcaggggctc aagatgcccc tgttctcatt tccgatcgcg acgatacaag
     4261 tcaggttgcc agctgccgca gcagcagcag tgcccagcac cacgagttct gcacaaggtc
     4321 ccccagtaaa atgatataca ttgacaccag tgaagatgcg gccgtcgcta gagagagctg
     4381 cgctggcgac gctgtagtct tcagagatgg ggatgctgtt gattgtagcc gttgctcttt
     4441 caatgagggt ggattcttct tgagacaaag gcttggccat ggtggcgcta gtgtcagaag
     4501 aatcgagctt tttgcaaaag cctaggcctc caaaaaagcc tcctcactac ttctggaata
     4561 gctcagaggc cgaggcggcc tcggcctctg cataaataaa aaaaattagt cagccatggg
     4621 gcggagaatg ggcggaactg ggcggagtta ggggcgggat gggcggagtt aggggcggga
     4681 ctatggttgc tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg
     4741 gggactttcc acacctggtt gctgactaat tgagatgcat gctttgcata cttctgcctg
     4801 ctggggagcc tggggacttt ccacacccta actgacacac attccacagc tggttctttc
     4861 cgcctcagaa ggtacctaac caagttcctc tttcagaggt tatttcaggc catggtgctg
     4921 cgc
//
