LOCUS       Exported                5730 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 5730)
  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..5730
                     /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..2442
                     /label=ARRB1
                     /note="Recieved from Kazusa in Nov 2009. Entered into 
                     database semi automatically by Ethan in Jan 2010. (note 
                     added in Ethan Strauss by 2/1/2010 10:30:20 AM"
     CDS             2452..2496
                     /codon_start=1
                     /label=15GS linker
                     /note="15GS linker"
                     /translation="GSSGGGGSGGGGSSG"
     CDS             2497..2529
                     /codon_start=1
                     /label=SmBiT
                     /note="SmBiT"
                     /translation="VTGYRLFEEIL"
     polyA_signal    2610..2831
                     /label=Sv40 late polyadenylation signal
     misc_feature    complement(2855..2888)
                     /label=Cre site
                     /note="Cre site"
     promoter        2897..2963
                     /label=EM7 bacterial promoter
     CDS             2977..3771
                     /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."
     misc_feature    3874..4796
                     /label=P15 ori
                     /note="P15 ori"
     misc_feature    4051
                     /label=C2129
                     /note="C2129"
     polyA_signal    complement(4818..4865)
                     /label=Synthetic PolyAdenylation Signal
     misc_feature    complement(4889..5287)
                     /label=BlastR
                     /note="BlastR"
     misc_feature    complement(5312..5730)
                     /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 gggcgacaaa
     1201 gggacccgag tgttcaagaa ggccagtcca aatggaaagc tcaccgtcta cctgggaaag
     1261 cgggactttg tggaccacat cgacctcgtg gaccctgtgg atggtgtggt cctggtggat
     1321 cctgagtatc tcaaagagcg gagagtctat gtgacgctga cctgcgcctt ccgctatggc
     1381 cgggaggacc tggatgtcct gggcctgacc tttcgcaagg acctgtttgt ggccaacgta
     1441 cagtcgttcc caccggcccc cgaggacaag aagcccctga cgcggctgca ggaacgcctc
     1501 atcaagaagc tgggcgagca cgcttaccct ttcacctttg agatccctcc aaaccttcca
     1561 tgttctgtga cactgcagcc ggggcccgaa gacacgggga aggcttgcgg tgtggactat
     1621 gaagtcaaag ccttctgcgc ggagaatttg gaggagaaga tccacaagcg gaattctgtg
     1681 cgtctggtca tccggaaggt tcagtatgcc ccagagaggc ctggccccca gcccacagcc
     1741 gagaccacca ggcagttcct catgtcggac aagcccttgc acctagaagc ctctctggat
     1801 aaggagatct attaccatgg agaacccatc agcgtcaacg tccacgtcac caacaacacc
     1861 aacaagacgg tgaagaagat caagatctca gtgcgccagt atgcagacat ctgccttttc
     1921 aacacagctc agtacaagtg ccctgttgcc atggaagagg ctgatgacac tgtggcaccc
     1981 agctcgacgt tctgcaaggt ctacacactg acccccttcc tagccaataa ccgagagaag
     2041 cggggcctcg ccttggacgg gaagctcaag cacgaagaca cgaacttggc ctctagcacc
     2101 ctgttgaggg aaggtgccaa ccgtgagatc ctggggatca ttgtttccta caaagtgaaa
     2161 gtgaagctgg tggtgtctcg gggcggcctg ttgggagatc ttgcatccag cgacgtggcc
     2221 gtggaactgc ccttcaccct aatgcacccc aagcccaaag aggaaccccc gcatcgggaa
     2281 gttccagaga acgagacgcc agtagatacc aatctcatag aacttgacac aaatgatgac
     2341 gacattgtat ttgaggactt tgctcgccag agactgaaag gcatgaagga tgacaaggag
     2401 gaagaggagg atggtaccgg ctctccacag ctcaacaaca gagtttctca aggcagttca
     2461 ggtggtggcg ggagcggagg tggaggctcg agcggtgtga ccggctaccg gctgttcgag
     2521 gagattctgt aaggccgcga ctctagagtc gacctgcagg catgcaagct gatttggctg
     2581 ctgccaccgc tgagcaaggc cgcttcgagc agacatgata agatacattg atgagtttgg
     2641 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     2701 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     2761 ttttatgttt caggttcagg gggagatgtg ggaggttttt ttaagcaagt aaaacctcta
     2821 caaatgtggt aaaatcttaa ttaagaattc taatataact tcgtatagca tacattatac
     2881 gaagttatgg atccactgtt gacaattaat catcggcata gtatatcggc atagtataat
     2941 acgacaaggt gaggaactaa acccaggagg cagatcatga ttgaacaaga tggattgcac
     3001 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     3061 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     3121 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     3181 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     3241 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     3301 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     3361 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     3421 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     3481 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     3541 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     3601 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     3661 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     3721 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg attcgaaatg
     3781 accgaccaag cgacgcccaa ccggtatcag ctcactcaaa ggcggtaata cggttatcca
     3841 cagaatcagg ggataacgca ggaaagaaca tgtcaaacat gagaattaca acttatatcg
     3901 tatggggctg acttcaggtg ctacatttga agagataaat tgcactgaaa tctagaaata
     3961 ttttatctga ttaataagat gatcttcttg agatcgtttt ggtctgcgcg taatctcttg
     4021 ctctgaaaac gaaaaaaccg ccttgcaggg cggtttttcg aaggttctct gagctaccaa
     4081 ctctttgaac cgaggtaact ggcttggagg agcgcagtca ccaaaacttg tcctttcagt
     4141 ttagccttaa ccggcgcatg acttcaagac taactcctct aaatcaatta ccagtggctg
     4201 ctgccagtgg tgcttttgca tgtctttccg ggttggactc aagacgatag ttaccggata
     4261 aggcgcagcg gtcggactga acggggggtt cgtgcataca gtccagcttg gagcgaactg
     4321 cctacccgga actgagtgtc aggcgtggaa tgagacaaac gcggccataa cagcggaatg
     4381 acaccggtaa accgaaaggc aggaacagga gagcgcacga gggagccgcc agggggaaac
     4441 gcctggtatc tttatagtcc tgtcgggttt cgccaccact gatttgagcg tcagatttcg
     4501 tgatgcttgt caggggggcg gagcctatgg aaaaacggct ttgccgcggc cctctcactt
     4561 ccctgttaag tatcttcctg gcatcttcca ggaaatctcc gccccgttcg taagccattt
     4621 ccgctcgccg cagtcgaacg accgagcgta gcgagtcagt gagcgaggaa gcggaatata
     4681 tcctgtatca catattctgc tgacgcaccg gtgcagcctt ttttctcctg ccacatgaag
     4741 cacttcactg acaccctcat cagtgccaac atagtaagcc agtatacact ccgctagcgc
     4801 tgatgtccgg aatcgtcaca caaaaaacca acacacagat gtaatgaaaa taaagatatt
     4861 ttattgcggc catcgtgatg gctagagttt agccctccca cacataacca gagggcagca
     4921 attcacgaat cccaactgcc gtcggctgtc catcactgtc cttcactatg gctttgatcc
     4981 caggatgcag atcgagaagc acctgtcggc accgtccgca ggggctcaag atgcccctgt
     5041 tctcatttcc gatcgcgacg atacaagtca ggttgccagc tgccgcagca gcagcagtgc
     5101 ccagcaccac gagttctgca caaggtcccc cagtaaaatg atatacattg acaccagtga
     5161 agatgcggcc gtcgctagag agagctgcgc tggcgacgct gtagtcttca gagatgggga
     5221 tgctgttgat tgtagccgtt gctctttcaa tgagggtgga ttcttcttga gacaaaggct
     5281 tggccatggt ggcgctagtg tcagaagaat cgagcttttt gcaaaagcct aggcctccaa
     5341 aaaagcctcc tcactacttc tggaatagct cagaggccga ggcggcctcg gcctctgcat
     5401 aaataaaaaa aattagtcag ccatggggcg gagaatgggc ggaactgggc ggagttaggg
     5461 gcgggatggg cggagttagg ggcgggacta tggttgctga ctaattgaga tgcatgcttt
     5521 gcatacttct gcctgctggg gagcctgggg actttccaca cctggttgct gactaattga
     5581 gatgcatgct ttgcatactt ctgcctgctg gggagcctgg ggactttcca caccctaact
     5641 gacacacatt ccacagctgg ttctttccgc ctcagaaggt acctaaccaa gttcctcttt
     5701 cagaggttat ttcaggccat ggtgctgcgc
//
