LOCUS       Exported                4670 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 4670)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Apr 15, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
COMMENT     pRSG195 designed by Brock F. Binkowski

            

            PCR amplify pF-BiBiTsmN 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..4670
                     /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
     CDS             1189..1224
                     /codon_start=1
                     /label=SmBiT
                     /note="SmBiT"
                     /translation="MVTGYRLFEEIL"
     CDS             1225..1269
                     /codon_start=1
                     /label=15GS linker
                     /note="15GS linker"
                     /translation="GSSGGGGSGGGGSSG"
     misc_feature    1270..1277
                     /label=SgfI Site
                     /note="Used in Flexi Vector Cloning"
     misc_feature    1279..1587
                     /label=Histone H4
                     /note="Recieved from Kazusa in August 2010. Entered into 
                     database semi automatically by Ethan in August 2010. 
                     Accession number NM_001034077 added 11/5/2010 from BLAST 
                     and length similarity. (note added in Automated Blaster by 
                     11/5/2010 4:49:09 PM"
     misc_feature    1588..1595
                     /label=PmeI site
                     /note="Used in Flexi Vector Cloning"
     polyA_signal    1724..1945
                     /label=Sv40 late polyadenylation signal
     promoter        2011..2077
                     /label=EM7 bacterial promoter
     CDS             2091..2885
                     /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      3040..3076
                     /direction=RIGHT
                     /label=ColE1-derived plasmid replication origin
                     /note="The actual origin is the first base."
     polyA_signal    complement(3758..3805)
                     /label=Synthetic PolyAdenylation Signal
     misc_feature    complement(3829..4227)
                     /label=BlastR
                     /note="BlastR"
     misc_feature    complement(4252..4670)
                     /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 cgctcaccat ggtgaccggc
     1201 taccggctgt tcgaggagat tctcgggagc tccggtggtg gcgggagcgg aggtggaggc
     1261 tcgagcggtg cgatcgccat gtccggcaga ggaaagggcg gaaaaggctt aggcaaaggg
     1321 ggcgctaagc gccaccgcaa ggtcttgaga gacaacattc agggcatcac caagcctgcc
     1381 attcggcgtc tagctcggcg tggcggcgtt aagcggatct ctggcctcat ttacgaggag
     1441 acccgcggtg tgctgaaggt gttcctggag aatgtgattc gggacgcagt cacctacacc
     1501 gagcacgcca agcgcaagac cgtcacagcc atggatgtgg tgtacgcgct caagcgccag
     1561 gggcgcaccc tgtacggctt cggaggcgtt taaacgaatt cgggctcggt acccggggat
     1621 cctctagagt cgacctgcag gcatgcaagc tgatccggct gctaacaaag cccgaaagga
     1681 agctgagttg gctgctgcca ccgctgagca aggccgcttc gagcagacat gataagatac
     1741 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     1801 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     1861 aacaattgca ttcattttat gtttcaggtt cagggggaga tgtgggaggt ttttttaagc
     1921 aagtaaaacc tctacaaatg tggtaaaatc ttaattaaga attctaatat aacttcgtat
     1981 agcatacatt atacgaagtt atggatccac tgttgacaat taatcatcgg catagtatat
     2041 cggcatagta taatacgaca aggtgaggaa ctaaacccag gaggcagatc atgattgaac
     2101 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     2161 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     2221 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
     2281 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     2341 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     2401 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     2461 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     2521 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     2581 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc
     2641 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     2701 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     2761 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     2821 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     2881 tctgattcga aatgaccgac caagcgacgc ccaaccggta tcagctcact caaaggcggt
     2941 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     3001 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     3061 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     3121 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     3181 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     3241 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     3301 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     3361 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     3421 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     3481 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     3541 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     3601 gcagattacg cgcagaaaaa aaggatttca agaagatcct ttgatctttt ctacggggtc
     3661 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     3721 gatcttcacc tagatccttt tatagtccgg aatcgtcaca caaaaaacca acacacagat
     3781 gtaatgaaaa taaagatatt ttattgcggc catcgtgatg gctagagttt agccctccca
     3841 cacataacca gagggcagca attcacgaat cccaactgcc gtcggctgtc catcactgtc
     3901 cttcactatg gctttgatcc caggatgcag atcgagaagc acctgtcggc accgtccgca
     3961 ggggctcaag atgcccctgt tctcatttcc gatcgcgacg atacaagtca ggttgccagc
     4021 tgccgcagca gcagcagtgc ccagcaccac gagttctgca caaggtcccc cagtaaaatg
     4081 atatacattg acaccagtga agatgcggcc gtcgctagag agagctgcgc tggcgacgct
     4141 gtagtcttca gagatgggga tgctgttgat tgtagccgtt gctctttcaa tgagggtgga
     4201 ttcttcttga gacaaaggct tggccatggt ggcgctagtg tcagaagaat cgagcttttt
     4261 gcaaaagcct aggcctccaa aaaagcctcc tcactacttc tggaatagct cagaggccga
     4321 ggcggcctcg gcctctgcat aaataaaaaa aattagtcag ccatggggcg gagaatgggc
     4381 ggaactgggc ggagttaggg gcgggatggg cggagttagg ggcgggacta tggttgctga
     4441 ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca
     4501 cctggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg
     4561 ggactttcca caccctaact gacacacatt ccacagctgg ttctttccgc ctcagaaggt
     4621 acctaaccaa gttcctcttt cagaggttat ttcaggccat ggtgctgcgc
//
