LOCUS       Exported                6878 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 6878)
  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..6878
                     /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..3621
                     /label=CTNNB1
                     /note="CTNNB1"
     misc_feature    3622..3629
                     /label=PmeI site
                     /note="Used in Flexi Vector Cloning"
     polyA_signal    3758..3979
                     /label=Sv40 late polyadenylation signal
     promoter        4045..4111
                     /label=EM7 bacterial promoter
     CDS             4125..4919
                     /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    5022..5944
                     /label=P15 ori
                     /note="P15 ori"
     misc_feature    5199
                     /label=C2129
                     /note="C2129"
     polyA_signal    complement(5966..6013)
                     /label=Synthetic PolyAdenylation Signal
     misc_feature    complement(6037..6435)
                     /label=BlastR
                     /note="BlastR"
     misc_feature    complement(6460..6878)
                     /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 ggctactcaa gctgatttga tggagttgga catggccatg
     1321 gaaccagaca gaaaagcggc tgttagtcac tggcagcaac agtcttacct ggactctgga
     1381 atccattctg gtgccactac cacagctcct tctctgagtg gtaaaggcaa tcctgaggaa
     1441 gaggatgtgg atacctccca agtcctgtat gagtgggaac agggattttc tcagtccttc
     1501 actcaagaac aagtagctga tattgatgga cagtatgcaa tgactcgagc tcagagggta
     1561 cgagctgcta tgttccctga gacattagat gagggcatgc agatcccatc tacacagttt
     1621 gatgctgctc atcccactaa tgtccagcgt ttggctgaac catcacagat gctgaaacat
     1681 gcagttgtaa acttgattaa ctatcaagat gatgcagaac ttgccacacg tgcaatccct
     1741 gaactgacaa aactgctaaa tgacgaggac caggtggtgg ttaataaggc tgcagttatg
     1801 gtccatcagc tttctaaaaa ggaagcttcc agacacgcta tcatgcgttc tcctcagatg
     1861 gtgtctgcta ttgtacgtac catgcagaat acaaatgatg tagaaacagc tcgttgtacc
     1921 gctgggacct tgcataacct ttcccatcat cgtgagggct tactggccat ctttaagtct
     1981 ggaggcattc ctgccctggt gaaaatgctt ggttcaccag tggattctgt gttgttttat
     2041 gccattacaa ctctccacaa ccttttatta catcaagaag gagctaaaat ggcagtgcgt
     2101 ttagctggtg ggctgcagaa aatggttgcc ttgctcaaca aaacaaatgt taaattcttg
     2161 gctattacga cagactgcct tcaaatttta gcttatggca accaagaaag caagctcatc
     2221 atactggcta gtggtggacc ccaagcttta gtaaatataa tgaggaccta tacttacgaa
     2281 aaactactgt ggaccacaag cagagtgctg aaggtgctat ctgtctgctc tagtaataag
     2341 ccggctattg tagaagctgg tggaatgcaa gctttaggac ttcacctgac agatccaagt
     2401 caacgtcttg ttcagaactg tctttggact ctcaggaatc tttcagatgc tgcaactaaa
     2461 caggaaggga tggaaggtct ccttgggact cttgttcagc ttctgggttc agatgatata
     2521 aatgtggtca cctgtgcagc tggaattctt tctaacctca cttgcaataa ttataagaac
     2581 aagatgatgg tctgccaagt gggtggtata gaggctcttg tgcgtactgt ccttcgggct
     2641 ggtgacaggg aagacatcac tgagcctgcc atctgtgctc ttcgtcatct gaccagccga
     2701 caccaagaag cagagatggc ccagaatgca gttcgccttc actatggact accagttgtg
     2761 gttaagctct tacacccacc atcccactgg cctctgataa aggctactgt tggattgatt
     2821 cgaaatcttg ccctttgtcc cgcaaatcat gcacctttgc gtgagcaggg tgccattcca
     2881 cgactagttc agttgcttgt tcgtgcacat caggataccc agcgccgtac gtccatgggt
     2941 gggacacagc agcaatttgt ggagggggtc cgcatggaag aaatagttga aggttgtacc
     3001 ggagcccttc acatcctagc tcgggatgtt cacaaccgaa ttgttatcag aggactaaat
     3061 accattccat tgtttgtgca gctgctttat tctcccattg aaaacatcca aagagtagct
     3121 gcaggggtcc tctgtgaact tgctcaggac aaggaagctg cagaagctat tgaagctgag
     3181 ggagccacag ctcctctgac agagttactt cactctagga atgaaggtgt ggcgacatat
     3241 gcagctgctg ttttgttccg aatgtctgag gacaagccac aagattacaa gaaacggctt
     3301 tcagttgagc tgaccagctc tctcttcaga acagagccaa tggcttggaa tgagactgct
     3361 gatcttggac ttgatattgg tgcccaggga gaaccccttg gatatcgcca ggatgatcct
     3421 agctatcgtt cttttcactc tggtggatat ggccaggatg ccttgggtat ggaccccatg
     3481 atggaacatg agatgggtgg ccaccaccct ggtgctgact atccagttga tgggctgcca
     3541 gatctggggc atgcccagga cctcatggat gggctgcctc caggtgacag caatcagctg
     3601 gcctggtttg atactgacct ggtttaaacg aattcgggct cggtacccgg ggatcctcta
     3661 gagtcgacct gcaggcatgc aagctgatcc ggctgctaac aaagcccgaa aggaagctga
     3721 gttggctgct gccaccgctg agcaaggccg cttcgagcag acatgataag atacattgat
     3781 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt
     3841 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat
     3901 tgcattcatt ttatgtttca ggttcagggg gagatgtggg aggttttttt aagcaagtaa
     3961 aacctctaca aatgtggtaa aatcttaatt aagaattcta atataacttc gtatagcata
     4021 cattatacga agttatggat ccactgttga caattaatca tcggcatagt atatcggcat
     4081 agtataatac gacaaggtga ggaactaaac ccaggaggca gatcatgatt gaacaagatg
     4141 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     4201 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     4261 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
     4321 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     4381 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     4441 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     4501 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     4561 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     4621 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg
     4681 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     4741 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     4801 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     4861 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgat
     4921 tcgaaatgac cgaccaagcg acgcccaacc ggtatcagct cactcaaagg cggtaatacg
     4981 gttatccaca gaatcagggg ataacgcagg aaagaacatg tcaaacatga gaattacaac
     5041 ttatatcgta tggggctgac ttcaggtgct acatttgaag agataaattg cactgaaatc
     5101 tagaaatatt ttatctgatt aataagatga tcttcttgag atcgttttgg tctgcgcgta
     5161 atctcttgct ctgaaaacga aaaaaccgcc ttgcagggcg gtttttcgaa ggttctctga
     5221 gctaccaact ctttgaaccg aggtaactgg cttggaggag cgcagtcacc aaaacttgtc
     5281 ctttcagttt agccttaacc ggcgcatgac ttcaagacta actcctctaa atcaattacc
     5341 agtggctgct gccagtggtg cttttgcatg tctttccggg ttggactcaa gacgatagtt
     5401 accggataag gcgcagcggt cggactgaac ggggggttcg tgcatacagt ccagcttgga
     5461 gcgaactgcc tacccggaac tgagtgtcag gcgtggaatg agacaaacgc ggccataaca
     5521 gcggaatgac accggtaaac cgaaaggcag gaacaggaga gcgcacgagg gagccgccag
     5581 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccaccactga tttgagcgtc
     5641 agatttcgtg atgcttgtca ggggggcgga gcctatggaa aaacggcttt gccgcggccc
     5701 tctcacttcc ctgttaagta tcttcctggc atcttccagg aaatctccgc cccgttcgta
     5761 agccatttcc gctcgccgca gtcgaacgac cgagcgtagc gagtcagtga gcgaggaagc
     5821 ggaatatatc ctgtatcaca tattctgctg acgcaccggt gcagcctttt ttctcctgcc
     5881 acatgaagca cttcactgac accctcatca gtgccaacat agtaagccag tatacactcc
     5941 gctagcgctg atgtccggaa tcgtcacaca aaaaaccaac acacagatgt aatgaaaata
     6001 aagatatttt attgcggcca tcgtgatggc tagagtttag ccctcccaca cataaccaga
     6061 gggcagcaat tcacgaatcc caactgccgt cggctgtcca tcactgtcct tcactatggc
     6121 tttgatccca ggatgcagat cgagaagcac ctgtcggcac cgtccgcagg ggctcaagat
     6181 gcccctgttc tcatttccga tcgcgacgat acaagtcagg ttgccagctg ccgcagcagc
     6241 agcagtgccc agcaccacga gttctgcaca aggtccccca gtaaaatgat atacattgac
     6301 accagtgaag atgcggccgt cgctagagag agctgcgctg gcgacgctgt agtcttcaga
     6361 gatggggatg ctgttgattg tagccgttgc tctttcaatg agggtggatt cttcttgaga
     6421 caaaggcttg gccatggtgg cgctagtgtc agaagaatcg agctttttgc aaaagcctag
     6481 gcctccaaaa aagcctcctc actacttctg gaatagctca gaggccgagg cggcctcggc
     6541 ctctgcataa ataaaaaaaa ttagtcagcc atggggcgga gaatgggcgg aactgggcgg
     6601 agttaggggc gggatgggcg gagttagggg cgggactatg gttgctgact aattgagatg
     6661 catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc tggttgctga
     6721 ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca
     6781 ccctaactga cacacattcc acagctggtt ctttccgcct cagaaggtac ctaaccaagt
     6841 tcctctttca gaggttattt caggccatgg tgctgcgc
//
