LOCUS       Exported                4955 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 4955)
  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..4955
                     /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..1872
                     /label=BCL2L11
                     /note="BCL2L11"
     misc_feature    1873..1880
                     /label=PmeI site
                     /note="Used in Flexi Vector Cloning"
     polyA_signal    2009..2230
                     /label=Sv40 late polyadenylation signal
     misc_feature    complement(2254..2287)
                     /label=Cre site
                     /note="Cre site"
     promoter        2296..2362
                     /label=EM7 bacterial promoter
     CDS             2376..3170
                     /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      3325..3361
                     /direction=RIGHT
                     /label=ColE1-derived plasmid replication origin
                     /note="The actual origin is the first base."
     polyA_signal    complement(4043..4090)
                     /label=Synthetic PolyAdenylation Signal
     misc_feature    complement(4114..4512)
                     /label=BlastR
                     /note="BlastR"
     misc_feature    complement(4537..4955)
                     /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 ggcaaagcaa ccttctgatg taagttctga gtgtgaccga
     1321 gaaggtagac aattgcagcc tgcggagagg cctccccagc tcagacctgg ggcccctacc
     1381 tccctacaga cagagccaca aggtaatcct gaaggcaatc acggaggtga aggggacagc
     1441 tgcccccacg gcagccctca gggcccgctg gccccacctg ccagccctgg cccttttgct
     1501 accagatccc cgcttttcat ctttatgaga agatcctccc tgctgtctcg atcctccagt
     1561 gggtatttct cttttgacac agacaggagc ccagcaccca tgagttgtga caaatcaaca
     1621 caaaccccaa gtcctccttg ccaggccttc aaccactatc tcagtgcaat ggcttccatg
     1681 aggcaggctg aacctgcaga tatgcgccca gagatatgga tcgcccaaga gttgcggcgt
     1741 attggagacg agtttaacgc ttactatgca aggagggtat ttttgaataa ttaccaagca
     1801 gccgaagacc acccacgaat ggttatctta cgactgttac gttacattgt ccgcctggtg
     1861 tggagaatgc atgtttaaac gaattcgggc tcggtacccg gggatcctct agagtcgacc
     1921 tgcaggcatg caagctgatc cggctgctaa caaagcccga aaggaagctg agttggctgc
     1981 tgccaccgct gagcaaggcc gcttcgagca gacatgataa gatacattga tgagtttgga
     2041 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
     2101 gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat
     2161 tttatgtttc aggttcaggg ggagatgtgg gaggtttttt taagcaagta aaacctctac
     2221 aaatgtggta aaatcttaat taagaattct aatataactt cgtatagcat acattatacg
     2281 aagttatgga tccactgttg acaattaatc atcggcatag tatatcggca tagtataata
     2341 cgacaaggtg aggaactaaa cccaggaggc agatcatgat tgaacaagat ggattgcacg
     2401 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     2461 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     2521 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt
     2581 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     2641 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     2701 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     2761 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     2821 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     2881 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg
     2941 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     3001 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     3061 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     3121 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga ttcgaaatga
     3181 ccgaccaagc gacgcccaac cggtatcagc tcactcaaag gcggtaatac ggttatccac
     3241 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     3301 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     3361 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     3421 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     3481 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     3541 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     3601 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     3661 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     3721 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     3781 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     3841 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     3901 aaaaaaagga tttcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     3961 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     4021 ccttttatag tccggaatcg tcacacaaaa aaccaacaca cagatgtaat gaaaataaag
     4081 atattttatt gcggccatcg tgatggctag agtttagccc tcccacacat aaccagaggg
     4141 cagcaattca cgaatcccaa ctgccgtcgg ctgtccatca ctgtccttca ctatggcttt
     4201 gatcccagga tgcagatcga gaagcacctg tcggcaccgt ccgcaggggc tcaagatgcc
     4261 cctgttctca tttccgatcg cgacgataca agtcaggttg ccagctgccg cagcagcagc
     4321 agtgcccagc accacgagtt ctgcacaagg tcccccagta aaatgatata cattgacacc
     4381 agtgaagatg cggccgtcgc tagagagagc tgcgctggcg acgctgtagt cttcagagat
     4441 ggggatgctg ttgattgtag ccgttgctct ttcaatgagg gtggattctt cttgagacaa
     4501 aggcttggcc atggtggcgc tagtgtcaga agaatcgagc tttttgcaaa agcctaggcc
     4561 tccaaaaaag cctcctcact acttctggaa tagctcagag gccgaggcgg cctcggcctc
     4621 tgcataaata aaaaaaatta gtcagccatg gggcggagaa tgggcggaac tgggcggagt
     4681 taggggcggg atgggcggag ttaggggcgg gactatggtt gctgactaat tgagatgcat
     4741 gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacctgg ttgctgacta
     4801 attgagatgc atgctttgca tacttctgcc tgctggggag cctggggact ttccacaccc
     4861 taactgacac acattccaca gctggttctt tccgcctcag aaggtaccta accaagttcc
     4921 tctttcagag gttatttcag gccatggtgc tgcgc
//
