LOCUS       Exported                5672 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 5672)
  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..5672
                     /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..2589
                     /label=VPS4A
                     /note="VPS4A"
     misc_feature    2590..2597
                     /label=PmeI site
                     /note="Used in Flexi Vector Cloning"
     polyA_signal    2726..2947
                     /label=Sv40 late polyadenylation signal
     promoter        3013..3079
                     /label=EM7 bacterial promoter
     CDS             3093..3887
                     /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      4042..4078
                     /direction=RIGHT
                     /label=ColE1-derived plasmid replication origin
                     /note="The actual origin is the first base."
     polyA_signal    complement(4760..4807)
                     /label=Synthetic PolyAdenylation Signal
     misc_feature    complement(4831..5229)
                     /label=BlastR
                     /note="BlastR"
     misc_feature    complement(5254..5672)
                     /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 gacaacgtca accctccaga aagccattga tctggtgacg
     1321 aaagccacag aggaggacaa agccaagaac tacgaggagg cgctgcggct gtaccagcat
     1381 gcggtggagt acttcctcca cgctatcaag tatgaggccc acagcgacaa ggccaaggag
     1441 agcattcgag ccaagtgcgt gcagtaccta gaccgggccg agaagctgaa ggattattta
     1501 cgaagcaaag agaaacacgg caagaagcca gtcaaagaga accagagtga gggcaagggc
     1561 agtgacagtg acagtgaagg ggataatccg gagaaaaaga aactgcaaga acagctgatg
     1621 ggtgccgtcg tgatggagaa gcccaacata cggtggaacg acgtggccgg gctggagggg
     1681 gccaaggagg ccctcaaaga agctgtcatt ttgccaatca aattcccaca cttgttcaca
     1741 ggcaagcgca ccccctggcg ggggattctg ctgttcggac cccctggcac agggaaatcc
     1801 tacctggcca aagccgtggc aacagaggcc aacaactcca ccttcttctc tgtgtcctcc
     1861 tcagatctga tgtccaagtg gctgggggag agtgaaaagc tggtcaagaa cctgtttgag
     1921 ctggccaggc agcacaagcc ctccatcatc ttcatcgatg aggtggattc cctctgcggg
     1981 tcccgaaatg aaaatgagag tgaggccgcc cggaggatca aaacggagtt cttggtccag
     2041 atgcaggggg tggggaataa caatgatggg actctggttc ttggagccac aaacatccca
     2101 tgggtgttgg attcggccat caggaggagg tttgaaaaac gaatttatat ccccttgccg
     2161 gaggaagctg cccgcgccca gatgttccgg ttgcatctcg ggagcactcc ccacaacctc
     2221 acggatgcaa acatccacga gctggcccgg aagacggaag gctactcggg cgcggacatc
     2281 agcatcatcg tgcgggactc tctcatgcag cccgtgagga aggtgcagtc ggccacacac
     2341 ttcaaaaagg tctgtggccc ctctcgcacc aaccccagca tgatgattga tgacctcctg
     2401 actccatgct caccagggga cccaggagcc atggagatga cttggatgga tgtccctggg
     2461 gacaaactct tagagcctgt ggtttgcatg tcggacatgc tgcggtctct ggccaccacc
     2521 cggcccacgg tgaatgcaga cgacctcctg aaagtgaaga aattctcaga ggactttggg
     2581 caagagagtg tttaaacgaa ttcgggctcg gtacccgggg atcctctaga gtcgacctgc
     2641 aggcatgcaa gctgatccgg ctgctaacaa agcccgaaag gaagctgagt tggctgctgc
     2701 caccgctgag caaggccgct tcgagcagac atgataagat acattgatga gtttggacaa
     2761 accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct
     2821 ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt
     2881 atgtttcagg ttcaggggga gatgtgggag gtttttttaa gcaagtaaaa cctctacaaa
     2941 tgtggtaaaa tcttaattaa gaattctaat ataacttcgt atagcataca ttatacgaag
     3001 ttatggatcc actgttgaca attaatcatc ggcatagtat atcggcatag tataatacga
     3061 caaggtgagg aactaaaccc aggaggcaga tcatgattga acaagatgga ttgcacgcag
     3121 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     3181 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     3241 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
     3301 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     3361 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     3421 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     3481 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     3541 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     3601 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg
     3661 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     3721 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     3781 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     3841 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgattc gaaatgaccg
     3901 accaagcgac gcccaaccgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     3961 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     4021 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     4081 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     4141 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     4201 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     4261 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     4321 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     4381 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     4441 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
     4501 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     4561 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     4621 aaaaggattt caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     4681 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     4741 tttatagtcc ggaatcgtca cacaaaaaac caacacacag atgtaatgaa aataaagata
     4801 ttttattgcg gccatcgtga tggctagagt ttagccctcc cacacataac cagagggcag
     4861 caattcacga atcccaactg ccgtcggctg tccatcactg tccttcacta tggctttgat
     4921 cccaggatgc agatcgagaa gcacctgtcg gcaccgtccg caggggctca agatgcccct
     4981 gttctcattt ccgatcgcga cgatacaagt caggttgcca gctgccgcag cagcagcagt
     5041 gcccagcacc acgagttctg cacaaggtcc cccagtaaaa tgatatacat tgacaccagt
     5101 gaagatgcgg ccgtcgctag agagagctgc gctggcgacg ctgtagtctt cagagatggg
     5161 gatgctgttg attgtagccg ttgctctttc aatgagggtg gattcttctt gagacaaagg
     5221 cttggccatg gtggcgctag tgtcagaaga atcgagcttt ttgcaaaagc ctaggcctcc
     5281 aaaaaagcct cctcactact tctggaatag ctcagaggcc gaggcggcct cggcctctgc
     5341 ataaataaaa aaaattagtc agccatgggg cggagaatgg gcggaactgg gcggagttag
     5401 gggcgggatg ggcggagtta ggggcgggac tatggttgct gactaattga gatgcatgct
     5461 ttgcatactt ctgcctgctg gggagcctgg ggactttcca cacctggttg ctgactaatt
     5521 gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc cacaccctaa
     5581 ctgacacaca ttccacagct ggttctttcc gcctcagaag gtacctaacc aagttcctct
     5641 ttcagaggtt atttcaggcc atggtgctgc gc
//
