LOCUS       Exported                4722 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 4722)
  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..4722
                     /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..1608
                     /label=CRaf RBD-CRD
                     /note="CRaf RBD-CRD"
     CDS             1618..1662
                     /codon_start=1
                     /label=15GS linker
                     /note="15GS linker"
                     /translation="GSSGGGGSGGGGSSG"
     CDS             1663..1695
                     /codon_start=1
                     /label=SmBiT
                     /note="SmBiT"
                     /translation="VTGYRLFEEIL"
     polyA_signal    1776..1997
                     /label=Sv40 late polyadenylation signal
     misc_feature    complement(2021..2054)
                     /label=Cre site
                     /note="Cre site"
     promoter        2063..2129
                     /label=EM7 bacterial promoter
     CDS             2143..2937
                     /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      3092..3128
                     /direction=RIGHT
                     /label=ColE1-derived plasmid replication origin
                     /note="The actual origin is the first base."
     polyA_signal    complement(3810..3857)
                     /label=Synthetic PolyAdenylation Signal
     misc_feature    complement(3881..4279)
                     /label=BlastR
                     /note="BlastR"
     misc_feature    complement(4304..4722)
                     /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 gccttctaag
     1201 acaagcaaca ctatccgtgt tttcttgccg aacaagcaaa gaacagtggt caatgtgcga
     1261 aatggaatga gcttgcatga ctgccttatg aaagcactca aggtgagggg cctgcaacca
     1321 gagtgctgtg cagtgttcag acttctccac gaacacaaag gtaaaaaagc acgcttagat
     1381 tggaatactg atgctgcgtc tttgattgga gaagaacttc aagtagattt cctggatcat
     1441 gttcccctca caacacacaa ctttgctcgg aagacgttcc tgaagcttgc cttctgtgac
     1501 atctgtcaga aattcctgct caatggattt cgatgtcaga cttgtggcta caaatttcat
     1561 gagcactgta gcaccaaagt acctactatg tgtgtggact ggagtaacgt ttctcaaggc
     1621 agttcaggtg gtggcgggag cggaggtgga ggctcgagcg gtgtgaccgg ctaccggctg
     1681 ttcgaggaga ttctgtaagg ccgcgactct agagtcgacc tgcaggcatg caagctgatt
     1741 tggctgctgc caccgctgag caaggccgct tcgagcagac atgataagat acattgatga
     1801 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     1861 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     1921 cattcatttt atgtttcagg ttcaggggga gatgtgggag gtttttttaa gcaagtaaaa
     1981 cctctacaaa tgtggtaaaa tcttaattaa gaattctaat ataacttcgt atagcataca
     2041 ttatacgaag ttatggatcc actgttgaca attaatcatc ggcatagtat atcggcatag
     2101 tataatacga caaggtgagg aactaaaccc aggaggcaga tcatgattga acaagatgga
     2161 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     2221 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     2281 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     2341 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     2401 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     2461 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     2521 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     2581 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     2641 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     2701 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     2761 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     2821 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     2881 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgattc
     2941 gaaatgaccg accaagcgac gcccaaccgg tatcagctca ctcaaaggcg gtaatacggt
     3001 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     3061 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     3121 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     3181 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     3241 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     3301 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     3361 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     3421 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     3481 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
     3541 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     3601 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     3661 cgcgcagaaa aaaaggattt caagaagatc ctttgatctt ttctacgggg tctgacgctc
     3721 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     3781 cctagatcct tttatagtcc ggaatcgtca cacaaaaaac caacacacag atgtaatgaa
     3841 aataaagata ttttattgcg gccatcgtga tggctagagt ttagccctcc cacacataac
     3901 cagagggcag caattcacga atcccaactg ccgtcggctg tccatcactg tccttcacta
     3961 tggctttgat cccaggatgc agatcgagaa gcacctgtcg gcaccgtccg caggggctca
     4021 agatgcccct gttctcattt ccgatcgcga cgatacaagt caggttgcca gctgccgcag
     4081 cagcagcagt gcccagcacc acgagttctg cacaaggtcc cccagtaaaa tgatatacat
     4141 tgacaccagt gaagatgcgg ccgtcgctag agagagctgc gctggcgacg ctgtagtctt
     4201 cagagatggg gatgctgttg attgtagccg ttgctctttc aatgagggtg gattcttctt
     4261 gagacaaagg cttggccatg gtggcgctag tgtcagaaga atcgagcttt ttgcaaaagc
     4321 ctaggcctcc aaaaaagcct cctcactact tctggaatag ctcagaggcc gaggcggcct
     4381 cggcctctgc ataaataaaa aaaattagtc agccatgggg cggagaatgg gcggaactgg
     4441 gcggagttag gggcgggatg ggcggagtta ggggcgggac tatggttgct gactaattga
     4501 gatgcatgct ttgcatactt ctgcctgctg gggagcctgg ggactttcca cacctggttg
     4561 ctgactaatt gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc
     4621 cacaccctaa ctgacacaca ttccacagct ggttctttcc gcctcagaag gtacctaacc
     4681 aagttcctct ttcagaggtt atttcaggcc atggtgctgc gc
//
