LOCUS       Exported                6324 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 6324)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Friday, May 30, 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..6324
                     /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"
     CDS             1189..3210
                     /label=PPP1R15A
     CDS             3220..3264
                     /codon_start=1
                     /label=15GS linker
                     /note="15GS linker"
                     /translation="GSSGGGGSGGGGSSG"
     CDS             3265..3297
                     /codon_start=1
                     /label=SmBiT
                     /note="SmBiT"
                     /translation="VTGYRLFEEIL"
     polyA_signal    3378..3599
                     /label=Sv40 late polyadenylation signal
     promoter        3665..3731
                     /label=EM7 bacterial promoter
     CDS             3745..4539
                     /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      4694..4730
                     /direction=RIGHT
                     /label=ColE1-derived plasmid replication origin
                     /note="The actual origin is the first base."
     polyA_signal    complement(5412..5459)
                     /label=Synthetic PolyAdenylation Signal
     misc_feature    complement(5483..5881)
                     /label=BlastR
                     /note="BlastR"
     misc_feature    complement(5906..6324)
                     /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 ggccccaggc
     1201 caagcacccc atcaggctac cccgtggagg gatgcccacc ctttcttcct cctgtcccca
     1261 gtgatgggcc tcctcagccg cgcctggagc cgcctgaggg gcctgggacc tctagagccc
     1321 tggctggtgg aagcagtaaa aggagcagct ctggtagaag ctggcctgga gggagaagct
     1381 aggactcctc tggcaatccc ccatacccct tggggcagac gccctgaaga ggaggctgaa
     1441 gacagtggag gccctggaga ggacagagaa acactggggc tgaaaaccag cagttccctt
     1501 cctgaagcct ggggactttt ggatgatgat gatggcatgt atggtgagcg agaggcaacc
     1561 agtgtcccta gagggcaggg aagtcaattt gcagatggcc agcgtgctcc cctgtctccc
     1621 agccttctga taaggacact gcaaggttct gataagaacc caggggagga gaaagccgag
     1681 gaagagggag ttgctgaaga ggagggagtt aacaagttct cttatccacc atcacaccgg
     1741 gagtgttgtc cagccgtgga ggaggaggac gatgaagaag ctgtaaagaa agaagctcac
     1801 agaacctcta cttctgcctt gtctccagga tccaagccca gcacttgggt gtcttgccca
     1861 ggggaggaag agaatcaagc cacggaggat aaaagaacag aaagaagtaa aggagccagg
     1921 aagacctccg tgtccccccg atcttcaggc tccgacccca ggtcctggga gtatcgttca
     1981 ggagaggcgt ccgaggagaa ggaggaaaag gcacacaaag aaactgggaa aggagaagct
     2041 gccccagggc cgcaatcctc agccccagcc cagaggcccc agctcaagtc ctggtggtgc
     2101 caacccagtg atgaagagga gggtgaggtc aaggctttgg gggcagctga gaaggatgga
     2161 gaagctgagt gtcctccctg catcccccca ccaagtgcct tcctgaaggc ctgggtgtat
     2221 tggccaggag aggacacaga ggaagaggaa gatgaggaag aagatgagga cagtgactct
     2281 ggatcagatg aggaagaggg agaagctgag gcttcctctt ccactcctgc tacaggtgtc
     2341 ttcttgaagt cctgggtcta tcagccagga gaggacacag aggaggagga agatgaggac
     2401 agtgatacag gatcagccga ggatgaaaga gaagctgaga cttctgcttc cacaccccct
     2461 gcaagtgctt tcttgaaggc ctgggtgtat cggccaggag aggacacgga ggaggaggaa
     2521 gatgaggatg tggatagtga ggataaggaa gatgattcag aagcagcctt gggagaagct
     2581 gagtcagacc cacatccctc ccacccggac cagagggccc acttcagggg ctggggatat
     2641 cgacctggaa aagagacaga ggaagaggaa gctgctgagg actggggaga agctgagccc
     2701 tgccccttcc gagtggccat ctatgtacct ggagagaagc caccgcctcc ctgggctcct
     2761 cctaggctgc ccctccgact gcaaaggcgg ctcaagcgcc cagaaacccc tactcatgat
     2821 ccggaccctg agactcccct aaaggccaga aaggtgcgct tctccgagaa ggtcactgtc
     2881 catttcctgg ctgtctgggc agggccggcc caggccgccc gccagggccc ctgggagcag
     2941 cttgctcggg atcgcagccg cttcgcacgc cgcatcaccc aggcccagga ggagctgagc
     3001 ccctgcctca cccctgctgc ccgggccaga gcctgggcac gcctcaggaa cccaccttta
     3061 gcccccatcc ctgccctcac ccagaccttg ccttcctcct ctgtcccttc gtccccagtc
     3121 cagaccacgc ccttgagcca agctgtggcc acaccttccc gctcgtctgc tgctgcagcg
     3181 gctgccctgg acctcagtgg gaggcgtggc gtttctcaag gcagttcagg tggtggcggg
     3241 agcggaggtg gaggctcgag cggtgtgacc ggctaccggc tgttcgagga gattctgtaa
     3301 ggccgcgact ctagagtcga cctgcaggca tgcaagctga tttggctgct gccaccgctg
     3361 agcaaggccg cttcgagcag acatgataag atacattgat gagtttggac aaaccacaac
     3421 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
     3481 aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca
     3541 ggttcagggg gagatgtggg aggttttttt aagcaagtaa aacctctaca aatgtggtaa
     3601 aatcttaatt aagaattcta atataacttc gtatagcata cattatacga agttatggat
     3661 ccactgttga caattaatca tcggcatagt atatcggcat agtataatac gacaaggtga
     3721 ggaactaaac ccaggaggca gatcatgatt gaacaagatg gattgcacgc aggttctccg
     3781 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     3841 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     3901 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     3961 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     4021 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     4081 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     4141 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     4201 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     4261 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     4321 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     4381 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     4441 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     4501 cgcatcgcct tctatcgcct tcttgacgag ttcttctgat tcgaaatgac cgaccaagcg
     4561 acgcccaacc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     4621 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     4681 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     4741 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     4801 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     4861 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     4921 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     4981 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     5041 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     5101 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     5161 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     5221 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     5281 ttcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     5341 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttatagt
     5401 ccggaatcgt cacacaaaaa accaacacac agatgtaatg aaaataaaga tattttattg
     5461 cggccatcgt gatggctaga gtttagccct cccacacata accagagggc agcaattcac
     5521 gaatcccaac tgccgtcggc tgtccatcac tgtccttcac tatggctttg atcccaggat
     5581 gcagatcgag aagcacctgt cggcaccgtc cgcaggggct caagatgccc ctgttctcat
     5641 ttccgatcgc gacgatacaa gtcaggttgc cagctgccgc agcagcagca gtgcccagca
     5701 ccacgagttc tgcacaaggt cccccagtaa aatgatatac attgacacca gtgaagatgc
     5761 ggccgtcgct agagagagct gcgctggcga cgctgtagtc ttcagagatg gggatgctgt
     5821 tgattgtagc cgttgctctt tcaatgaggg tggattcttc ttgagacaaa ggcttggcca
     5881 tggtggcgct agtgtcagaa gaatcgagct ttttgcaaaa gcctaggcct ccaaaaaagc
     5941 ctcctcacta cttctggaat agctcagagg ccgaggcggc ctcggcctct gcataaataa
     6001 aaaaaattag tcagccatgg ggcggagaat gggcggaact gggcggagtt aggggcggga
     6061 tgggcggagt taggggcggg actatggttg ctgactaatt gagatgcatg ctttgcatac
     6121 ttctgcctgc tggggagcct ggggactttc cacacctggt tgctgactaa ttgagatgca
     6181 tgctttgcat acttctgcct gctggggagc ctggggactt tccacaccct aactgacaca
     6241 cattccacag ctggttcttt ccgcctcaga aggtacctaa ccaagttcct ctttcagagg
     6301 ttatttcagg ccatggtgct gcgc
//
