LOCUS       Exported                5664 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 5664)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Apr 15, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
COMMENT     pRSG194 designed by Brock F. Binkowski

            

            PCR amplify pF-BiBiTsmC 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..5664
                     /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
     misc_feature    1180..1187
                     /label=SgfI Site
                     /note="Used in Flexi Vector Cloning"
     misc_feature    1189..2376
                     /label=SLC35A2 (co)
                     /note="SLC35A2 (co)"
     CDS             2386..2430
                     /codon_start=1
                     /label=15GS linker
                     /note="15GS linker"
                     /translation="GSSGGGGSGGGGSSG"
     CDS             2431..2463
                     /codon_start=1
                     /label=SmBiT
                     /note="SmBiT"
                     /translation="VTGYRLFEEIL"
     polyA_signal    2544..2765
                     /label=Sv40 late polyadenylation signal
     misc_feature    complement(2789..2822)
                     /label=Cre site
                     /note="Cre site"
     promoter        2831..2897
                     /label=EM7 bacterial promoter
     CDS             2911..3705
                     /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    3808..4730
                     /label=P15 ori
                     /note="P15 ori"
     misc_feature    3985
                     /label=C2129
                     /note="C2129"
     polyA_signal    complement(4752..4799)
                     /label=Synthetic PolyAdenylation Signal
     misc_feature    complement(4823..5221)
                     /label=BlastR
                     /note="BlastR"
     misc_feature    complement(5246..5664)
                     /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 cgatcgccat ggctgccgtg
     1201 ggcgcaggag gaagtacggc agcacctggt ccgggcgcgg tgtctgcagg cgctcttgaa
     1261 cctgggactg cttcagccgc ccaccgaaga ttgaagtata tttctcttgc tgttctcgta
     1321 gtgcaaaatg cttccctcat cctttctata agatacgctc ggacgttgcc gggcgatcga
     1381 tttttcgcaa caaccgcagt cgtcatggca gaggtcctta agggattgac gtgcctgctc
     1441 cttctctttg cgcaaaaacg aggaaatgta aaacacctgg ttctgtttct gcacgaggca
     1501 gtgcttgtcc aatatgttga cacactgaag ttggcggtac ccagcctcat ttatactttg
     1561 caaaataatc ttcaatatgt cgctatatca aatttgccgg cagctacgtt ccaagtcacc
     1621 taccaactga aaatcctgac aacggcattg ttctccgttc ttatgctgaa ccggtccttg
     1681 tcccgactcc aatgggccag tctccttctc ctctttacag gggtcgccat agttcaagct
     1741 caacaagcgg gaggaggagg tccccgccct ctcgatcaga atccgggagc cgggctcgcg
     1801 gctgtggtgg cctcctgcct tagctctggg ttcgcgggtg tatattttga gaagatcctg
     1861 aagggttctt ctggtagtgt ctggttgaga aatctgcagc ttggcctctt tgggacggcc
     1921 ctcgggttgg tgggtttgtg gtgggcggag ggcaccgcag tcgcaactcg cggattcttt
     1981 tttggataca caccagctgt ttggggtgtc gtattgaacc aagcatttgg cggattgctc
     2041 gtggcggttg ttgtgaaata cgcggacaac atactgaagg gattcgcgac ctccttgtcc
     2101 atcgtattgt ccaccgtcgc ctcaataaga ctcttcgggt ttcatgtaga tccgctcttt
     2161 gccttgggtg ctgggcttgt cataggagcg gtctatttgt actccctgcc tcggggagca
     2221 gcgaaagcca tagcctctgc ttccgcgtct gcctccggtc cgtgcgtcca tcagcaacct
     2281 ccgggacagc cgccgcctcc acagttgtcc tcccaccgcg gtgacctcat aaccgagccc
     2341 ttcctgccga aattgctgac caaagtgaag ggatccgttt ctcaaggcag ttcaggtggt
     2401 ggcgggagcg gaggtggagg ctcgagcggt gtgaccggct accggctgtt cgaggagatt
     2461 ctgtaaggcc gcgactctag agtcgacctg caggcatgca agctgatttg gctgctgcca
     2521 ccgctgagca aggccgcttc gagcagacat gataagatac attgatgagt ttggacaaac
     2581 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     2641 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     2701 gtttcaggtt cagggggaga tgtgggaggt ttttttaagc aagtaaaacc tctacaaatg
     2761 tggtaaaatc ttaattaaga attctaatat aacttcgtat agcatacatt atacgaagtt
     2821 atggatccac tgttgacaat taatcatcgg catagtatat cggcatagta taatacgaca
     2881 aggtgaggaa ctaaacccag gaggcagatc atgattgaac aagatggatt gcacgcaggt
     2941 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     3001 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     3061 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     3121 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     3181 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     3241 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     3301 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     3361 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     3421 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     3481 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     3541 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     3601 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     3661 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgattcga aatgaccgac
     3721 caagcgacgc ccaaccggta tcagctcact caaaggcggt aatacggtta tccacagaat
     3781 caggggataa cgcaggaaag aacatgtcaa acatgagaat tacaacttat atcgtatggg
     3841 gctgacttca ggtgctacat ttgaagagat aaattgcact gaaatctaga aatattttat
     3901 ctgattaata agatgatctt cttgagatcg ttttggtctg cgcgtaatct cttgctctga
     3961 aaacgaaaaa accgccttgc agggcggttt ttcgaaggtt ctctgagcta ccaactcttt
     4021 gaaccgaggt aactggcttg gaggagcgca gtcaccaaaa cttgtccttt cagtttagcc
     4081 ttaaccggcg catgacttca agactaactc ctctaaatca attaccagtg gctgctgcca
     4141 gtggtgcttt tgcatgtctt tccgggttgg actcaagacg atagttaccg gataaggcgc
     4201 agcggtcgga ctgaacgggg ggttcgtgca tacagtccag cttggagcga actgcctacc
     4261 cggaactgag tgtcaggcgt ggaatgagac aaacgcggcc ataacagcgg aatgacaccg
     4321 gtaaaccgaa aggcaggaac aggagagcgc acgagggagc cgccaggggg aaacgcctgg
     4381 tatctttata gtcctgtcgg gtttcgccac cactgatttg agcgtcagat ttcgtgatgc
     4441 ttgtcagggg ggcggagcct atggaaaaac ggctttgccg cggccctctc acttccctgt
     4501 taagtatctt cctggcatct tccaggaaat ctccgccccg ttcgtaagcc atttccgctc
     4561 gccgcagtcg aacgaccgag cgtagcgagt cagtgagcga ggaagcggaa tatatcctgt
     4621 atcacatatt ctgctgacgc accggtgcag ccttttttct cctgccacat gaagcacttc
     4681 actgacaccc tcatcagtgc caacatagta agccagtata cactccgcta gcgctgatgt
     4741 ccggaatcgt cacacaaaaa accaacacac agatgtaatg aaaataaaga tattttattg
     4801 cggccatcgt gatggctaga gtttagccct cccacacata accagagggc agcaattcac
     4861 gaatcccaac tgccgtcggc tgtccatcac tgtccttcac tatggctttg atcccaggat
     4921 gcagatcgag aagcacctgt cggcaccgtc cgcaggggct caagatgccc ctgttctcat
     4981 ttccgatcgc gacgatacaa gtcaggttgc cagctgccgc agcagcagca gtgcccagca
     5041 ccacgagttc tgcacaaggt cccccagtaa aatgatatac attgacacca gtgaagatgc
     5101 ggccgtcgct agagagagct gcgctggcga cgctgtagtc ttcagagatg gggatgctgt
     5161 tgattgtagc cgttgctctt tcaatgaggg tggattcttc ttgagacaaa ggcttggcca
     5221 tggtggcgct agtgtcagaa gaatcgagct ttttgcaaaa gcctaggcct ccaaaaaagc
     5281 ctcctcacta cttctggaat agctcagagg ccgaggcggc ctcggcctct gcataaataa
     5341 aaaaaattag tcagccatgg ggcggagaat gggcggaact gggcggagtt aggggcggga
     5401 tgggcggagt taggggcggg actatggttg ctgactaatt gagatgcatg ctttgcatac
     5461 ttctgcctgc tggggagcct ggggactttc cacacctggt tgctgactaa ttgagatgca
     5521 tgctttgcat acttctgcct gctggggagc ctggggactt tccacaccct aactgacaca
     5581 cattccacag ctggttcttt ccgcctcaga aggtacctaa ccaagttcct ctttcagagg
     5641 ttatttcagg ccatggtgct gcgc
//
