LOCUS       Exported                6440 bp ds-DNA     circular SYN 30-MAY-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6440)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Friday, May 30, 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..6440
                     /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..3183
                     /label=SYK
                     /note="SYK"
     misc_feature    3184..3191
                     /label=PmeI site
                     /note="Used in Flexi Vector Cloning"
     polyA_signal    3320..3541
                     /label=Sv40 late polyadenylation signal
     misc_feature    complement(3565..3598)
                     /label=Cre site
                     /note="Cre site"
     promoter        3607..3673
                     /label=EM7 bacterial promoter
     CDS             3687..4481
                     /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    4584..5506
                     /label=P15 ori
                     /note="P15 ori"
     misc_feature    4761
                     /label=C2129
                     /note="C2129"
     polyA_signal    complement(5528..5575)
                     /label=Synthetic PolyAdenylation Signal
     misc_feature    complement(5599..5997)
                     /label=BlastR
                     /note="BlastR"
     misc_feature    complement(6022..6440)
                     /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 ggccagcagc ggcatggctg acagcgccaa ccacctgccc
     1321 ttctttttcg gcaacatcac ccgggaggag gcagaagatt acctggtcca ggggggcatg
     1381 agtgatgggc tttatttgct gcgccagagc cgcaactacc tgggtggctt cgccctgtcc
     1441 gtggcccacg ggaggaaggc acaccactac accatcgagc gggagctgaa tggcacctac
     1501 gccatcgccg gtggcaggac ccatgccagc cccgccgacc tctgccacta ccactcccag
     1561 gagtctgatg gcctggtctg cctcctcaag aagcccttca accggcccca aggggtgcag
     1621 cccaagactg ggccctttga ggatttgaag gaaaacctca tcagggaata tgtgaagcag
     1681 acatggaacc tgcagggtca ggctctggag caggccatca tcagtcagaa gcctcagctg
     1741 gagaagctga tcgctaccac agcccatgaa aaaatgcctt ggttccatgg aaaaatctct
     1801 cgggaagaat ctgagcaaat tgtcctgata ggatcaaaga caaatggaaa gttcctgatc
     1861 cgagccagag acaacaacgg ctcctacgcc ctgtgcctgc tgcacgaagg gaaggtgctg
     1921 cactatcgca tcgacaaaga caagacaggg aagctctcca tccccgaggg aaagaagttc
     1981 gacacgctct ggcagctagt cgagcattat tcttataaag cagatggttt gttaagagtt
     2041 cttactgtcc catgtcaaaa aatcggcaca cagggaaatg ttaattttgg aggccgtcca
     2101 caacttccag gttcccatcc tgcgacttgg tcagcgggtg gaataatctc aagaatcaaa
     2161 tcatactcct tcccaaagcc tggccacaga aagtcctccc ctgcccaagg gaaccggcaa
     2221 gagagtactg tgtcattcaa tccgtatgag ccagaacttg caccctgggc tgcagacaaa
     2281 ggcccccaga gagaagccct acccatggac acagaggtgt acgagagccc ctacgcggac
     2341 cccgaggaga tcaggcccaa ggaggtttac ctggaccgaa agctgctgac gctggaagac
     2401 aaagaactgg gctctggtaa ttttggaact gtgaaaaagg gctactacca aatgaaaaaa
     2461 gttgtgaaaa ccgtggctgt gaaaatactg aaaaacgagg ccaatgaccc cgctcttaaa
     2521 gatgagttat tagcagaagc aaatgtcatg cagcagctgg acaacccgta catcgtgcgg
     2581 atgatcggga tatgcgaggc cgagtcctgg atgctggtta tggagatggc agaacttggt
     2641 cccctcaata agtatttgca gcagaacaga catgtcaagg ataagaacat catagaactg
     2701 gttcatcagg tttccatggg catgaagtac ttggaggaga gcaattttgt gcacagagat
     2761 ctggctgcaa gaaatgtgtt gctagttacc caacattacg ccaagatcag tgatttcgga
     2821 ctttccaaag cactgcgtgc tgatgaaaac tactacaagg cccagaccca tggaaagtgg
     2881 cctgtcaagt ggtacgctcc ggaatgcatc aactactaca agttctccag caaaagcgat
     2941 gtctggagct ttggagtgtt gatgtgggaa gcattctcct atgggcagaa gccatatcga
     3001 gggatgaaag gaagtgaagt caccgctatg ttagagaaag gagagcggat ggggtgccct
     3061 gcagggtgtc caagagagat gtacgatctc atgaatctgt gctggacata cgatgtggaa
     3121 aacaggcccg gattcgcagc agtggaactg cggctgcgca attactacta tgacgtggtg
     3181 aacgtttaaa cgaattcggg ctcggtaccc ggggatcctc tagagtcgac ctgcaggcat
     3241 gcaagctgat ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc
     3301 tgagcaaggc cgcttcgagc agacatgata agatacattg atgagtttgg acaaaccaca
     3361 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt
     3421 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt
     3481 caggttcagg gggagatgtg ggaggttttt ttaagcaagt aaaacctcta caaatgtggt
     3541 aaaatcttaa ttaagaattc taatataact tcgtatagca tacattatac gaagttatgg
     3601 atccactgtt gacaattaat catcggcata gtatatcggc atagtataat acgacaaggt
     3661 gaggaactaa acccaggagg cagatcatga ttgaacaaga tggattgcac gcaggttctc
     3721 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     3781 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     3841 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca
     3901 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     3961 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     4021 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     4081 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     4141 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     4201 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     4261 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     4321 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     4381 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     4441 agcgcatcgc cttctatcgc cttcttgacg agttcttctg attcgaaatg accgaccaag
     4501 cgacgcccaa ccggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     4561 ggataacgca ggaaagaaca tgtcaaacat gagaattaca acttatatcg tatggggctg
     4621 acttcaggtg ctacatttga agagataaat tgcactgaaa tctagaaata ttttatctga
     4681 ttaataagat gatcttcttg agatcgtttt ggtctgcgcg taatctcttg ctctgaaaac
     4741 gaaaaaaccg ccttgcaggg cggtttttcg aaggttctct gagctaccaa ctctttgaac
     4801 cgaggtaact ggcttggagg agcgcagtca ccaaaacttg tcctttcagt ttagccttaa
     4861 ccggcgcatg acttcaagac taactcctct aaatcaatta ccagtggctg ctgccagtgg
     4921 tgcttttgca tgtctttccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     4981 gtcggactga acggggggtt cgtgcataca gtccagcttg gagcgaactg cctacccgga
     5041 actgagtgtc aggcgtggaa tgagacaaac gcggccataa cagcggaatg acaccggtaa
     5101 accgaaaggc aggaacagga gagcgcacga gggagccgcc agggggaaac gcctggtatc
     5161 tttatagtcc tgtcgggttt cgccaccact gatttgagcg tcagatttcg tgatgcttgt
     5221 caggggggcg gagcctatgg aaaaacggct ttgccgcggc cctctcactt ccctgttaag
     5281 tatcttcctg gcatcttcca ggaaatctcc gccccgttcg taagccattt ccgctcgccg
     5341 cagtcgaacg accgagcgta gcgagtcagt gagcgaggaa gcggaatata tcctgtatca
     5401 catattctgc tgacgcaccg gtgcagcctt ttttctcctg ccacatgaag cacttcactg
     5461 acaccctcat cagtgccaac atagtaagcc agtatacact ccgctagcgc tgatgtccgg
     5521 aatcgtcaca caaaaaacca acacacagat gtaatgaaaa taaagatatt ttattgcggc
     5581 catcgtgatg gctagagttt agccctccca cacataacca gagggcagca attcacgaat
     5641 cccaactgcc gtcggctgtc catcactgtc cttcactatg gctttgatcc caggatgcag
     5701 atcgagaagc acctgtcggc accgtccgca ggggctcaag atgcccctgt tctcatttcc
     5761 gatcgcgacg atacaagtca ggttgccagc tgccgcagca gcagcagtgc ccagcaccac
     5821 gagttctgca caaggtcccc cagtaaaatg atatacattg acaccagtga agatgcggcc
     5881 gtcgctagag agagctgcgc tggcgacgct gtagtcttca gagatgggga tgctgttgat
     5941 tgtagccgtt gctctttcaa tgagggtgga ttcttcttga gacaaaggct tggccatggt
     6001 ggcgctagtg tcagaagaat cgagcttttt gcaaaagcct aggcctccaa aaaagcctcc
     6061 tcactacttc tggaatagct cagaggccga ggcggcctcg gcctctgcat aaataaaaaa
     6121 aattagtcag ccatggggcg gagaatgggc ggaactgggc ggagttaggg gcgggatggg
     6181 cggagttagg ggcgggacta tggttgctga ctaattgaga tgcatgcttt gcatacttct
     6241 gcctgctggg gagcctgggg actttccaca cctggttgct gactaattga gatgcatgct
     6301 ttgcatactt ctgcctgctg gggagcctgg ggactttcca caccctaact gacacacatt
     6361 ccacagctgg ttctttccgc ctcagaaggt acctaaccaa gttcctcttt cagaggttat
     6421 ttcaggccat ggtgctgcgc
//
