LOCUS       Exported                5433 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 5433)
  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..5433
                     /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..2319
                     /label=NCK1
                     /note="NCK1"
     misc_feature    1192..1371
                     /label=SH1
                     /note="SH1"
     CDS             2329..2373
                     /codon_start=1
                     /label=15GS linker
                     /note="15GS linker"
                     /translation="GSSGGGGSGGGGSSG"
     CDS             2374..2406
                     /codon_start=1
                     /label=SmBiT
                     /note="SmBiT"
                     /translation="VTGYRLFEEIL"
     polyA_signal    2487..2708
                     /label=Sv40 late polyadenylation signal
     misc_feature    complement(2732..2765)
                     /label=Cre site
                     /note="Cre site"
     promoter        2774..2840
                     /label=EM7 bacterial promoter
     CDS             2854..3648
                     /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      3803..3839
                     /direction=RIGHT
                     /label=ColE1-derived plasmid replication origin
                     /note="The actual origin is the first base."
     polyA_signal    complement(4521..4568)
                     /label=Synthetic PolyAdenylation Signal
     misc_feature    complement(4592..4990)
                     /label=BlastR
                     /note="BlastR"
     misc_feature    complement(5015..5433)
                     /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 ggcagaagaa
     1201 gtggtggtag tagccaaatt tgattatgtg gcccaacaag aacaagagtt ggacatcaag
     1261 aagaatgaga gattatggct tctggatgat tctaagtcct ggtggcgagt tcgaaattcc
     1321 atgaataaaa caggttttgt gccttctaac tatgtggaaa ggaaaaacag tgctcggaaa
     1381 gcatctattg tgaaaaacct aaaggatacc ttaggcattg gaaaagtgaa aagaaaacct
     1441 agtgtgccag attctgcatc tcctgctgat gatagttttg ttgacccagg ggaacgtctc
     1501 tatgacctca acatgcccgc ttatgtgaaa tttaactaca tggctgagag agaggatgaa
     1561 ttatcattga taaaggggac aaaggtgatc gtcatggaga aatgcagtga tgggtggtgg
     1621 cgtggtagct acaatggaca agttggatgg ttcccttcaa actatgtaac tgaagaaggt
     1681 gacagtcctt tgggtgacca tgtgggttct ctgtcagaga aattagcagc agtcgtcaat
     1741 aacctaaata ctgggcaagt gttgcatgtg gtacaggctc tttacccatt cagctcatct
     1801 aatgatgaag aacttaattt cgagaaagga gatgtaatgg atgttattga aaaacctgaa
     1861 aatgacccag agtggtggaa atgcaggaag atcaatggta tggttggtct agtaccaaaa
     1921 aactatgtta ccgttatgca gaataatcca ttaacttcag gtttggaacc atcacctcca
     1981 cagtgtgatt acattaggcc ttcactcact ggaaagtttg ctggcaatcc ttggtattat
     2041 ggcaaagtca ccaggcatca agcagaaatg gcattaaatg aaagaggaca tgaaggggat
     2101 ttcctcattc gtgatagtga atcttcgcca aatgatttct cagtatcact aaaagcacaa
     2161 gggaaaaaca agcattttaa agtccaacta aaagagactg tctactgcat tgggcagcgt
     2221 aaattcagca ccatggaaga acttgtagaa cattacaaaa aggcaccaat ttttacaagt
     2281 gaacaaggag aaaaattata tcttgtcaag catttatcag tttctcaagg cagttcaggt
     2341 ggtggcggga gcggaggtgg aggctcgagc ggtgtgaccg gctaccggct gttcgaggag
     2401 attctgtaag gccgcgactc tagagtcgac ctgcaggcat gcaagctgat ttggctgctg
     2461 ccaccgctga gcaaggccgc ttcgagcaga catgataaga tacattgatg agtttggaca
     2521 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     2581 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
     2641 tatgtttcag gttcaggggg agatgtggga ggttttttta agcaagtaaa acctctacaa
     2701 atgtggtaaa atcttaatta agaattctaa tataacttcg tatagcatac attatacgaa
     2761 gttatggatc cactgttgac aattaatcat cggcatagta tatcggcata gtataatacg
     2821 acaaggtgag gaactaaacc caggaggcag atcatgattg aacaagatgg attgcacgca
     2881 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     2941 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     3001 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
     3061 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     3121 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     3181 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     3241 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     3301 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     3361 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc
     3421 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     3481 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     3541 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     3601 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgatt cgaaatgacc
     3661 gaccaagcga cgcccaaccg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     3721 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     3781 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     3841 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     3901 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     3961 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     4021 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     4081 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     4141 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     4201 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     4261 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     4321 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     4381 aaaaaggatt tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     4441 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     4501 ttttatagtc cggaatcgtc acacaaaaaa ccaacacaca gatgtaatga aaataaagat
     4561 attttattgc ggccatcgtg atggctagag tttagccctc ccacacataa ccagagggca
     4621 gcaattcacg aatcccaact gccgtcggct gtccatcact gtccttcact atggctttga
     4681 tcccaggatg cagatcgaga agcacctgtc ggcaccgtcc gcaggggctc aagatgcccc
     4741 tgttctcatt tccgatcgcg acgatacaag tcaggttgcc agctgccgca gcagcagcag
     4801 tgcccagcac cacgagttct gcacaaggtc ccccagtaaa atgatataca ttgacaccag
     4861 tgaagatgcg gccgtcgcta gagagagctg cgctggcgac gctgtagtct tcagagatgg
     4921 ggatgctgtt gattgtagcc gttgctcttt caatgagggt ggattcttct tgagacaaag
     4981 gcttggccat ggtggcgcta gtgtcagaag aatcgagctt tttgcaaaag cctaggcctc
     5041 caaaaaagcc tcctcactac ttctggaata gctcagaggc cgaggcggcc tcggcctctg
     5101 cataaataaa aaaaattagt cagccatggg gcggagaatg ggcggaactg ggcggagtta
     5161 ggggcgggat gggcggagtt aggggcggga ctatggttgc tgactaattg agatgcatgc
     5221 tttgcatact tctgcctgct ggggagcctg gggactttcc acacctggtt gctgactaat
     5281 tgagatgcat gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacccta
     5341 actgacacac attccacagc tggttctttc cgcctcagaa ggtacctaac caagttcctc
     5401 tttcagaggt tatttcaggc catggtgctg cgc
//
