LOCUS       Exported                6219 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 6219)
  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..6219
                     /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..2931
                     /label=PINK1
                     /note="This insert contains an Sgf I site at position 0 of 
                     the unmodified insert. This insert contains a Pme I site at
                     position 1752 of the unmodified insert."
     CDS             2941..2985
                     /codon_start=1
                     /label=15GS linker
                     /note="15GS linker"
                     /translation="GSSGGGGSGGGGSSG"
     CDS             2986..3018
                     /codon_start=1
                     /label=SmBiT
                     /note="SmBiT"
                     /translation="VTGYRLFEEIL"
     polyA_signal    3099..3320
                     /label=Sv40 late polyadenylation signal
     promoter        3386..3452
                     /label=EM7 bacterial promoter
     CDS             3466..4260
                     /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    4363..5285
                     /label=P15 ori
                     /note="P15 ori"
     misc_feature    4540
                     /label=C2129
                     /note="C2129"
     polyA_signal    complement(5307..5354)
                     /label=Synthetic PolyAdenylation Signal
     misc_feature    complement(5378..5776)
                     /label=BlastR
                     /note="BlastR"
     misc_feature    complement(5801..6219)
                     /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 ggccgtgagg
     1201 caggccctgg gaaggggact gcagctggga cgggccctgc tgctgagatt cacaggcaag
     1261 cctggcaggg cctacggact gggccgccca ggaccagcag caggctgcgt gcggggcgag
     1321 agacctggct gggccgccgg ccctggcgcc gagccacgga gagtgggact gggactgcca
     1381 aacaggctgc gcttctttag gcagtctgtg gcaggactgg cagcaaggct gcagaggcag
     1441 tttgtggtgc gggcatgggg atgcgcagga ccatgtggca gagccgtgtt cctggccttt
     1501 ggcctgggcc tgggcctgat cgaggagaag caggcagagt ctaggagggc cgtgagcgcc
     1561 tgccaggaga tccaggccat cttcacacag aagagcaagc ctggaccaga cccactggat
     1621 acccggagac tgcagggctt tcgcctggag gagtacctga tcggccagtc catcggcaag
     1681 ggctgttctg ccgccgtgta tgaggccacc atgcctacac tgccacagaa cctggaggtg
     1741 accaagtcta caggcctgct gccaggaagg ggacctggca ccagcgcccc cggcgagggc
     1801 caggagagag cccccggcgc ccctgccttc ccactggcca tcaagatgat gtggaacatc
     1861 agcgccggca gctcctctga ggccatcctg aatacaatga gccaggagct ggtgccagca
     1921 tccagggtgg ccctggcagg cgagtacggc gccgtgacct ataggaagtc caagagggga
     1981 cctaagcagc tggcacctca cccaaatatc atccgggtgc tgagagcctt cacaagctcc
     2041 gtgccactgc tgcctggcgc cctggtggac tacccagatg tgctgccctc caggctgcac
     2101 ccagagggac tgggacacgg acggaccctg ttcctggtca tgaagaacta cccttgcaca
     2161 ctgcggcagt acctgtgcgt gaataccccc tctcctagac tggccgccat gatgctgctg
     2221 cagctgctgg agggagtgga ccacctggtg cagcagggaa tcgcacaccg ggacctgaag
     2281 tctgataata tcctggtgga gctggaccct gatggctgcc catggctggt catcgccgac
     2341 ttcggctgct gtctggccga tgagagcatc ggcctgcagc tgccattttc tagctggtac
     2401 gtggatagag gaggaaacgg atgtctgatg gcaccagagg tgtccacagc aaggccaggc
     2461 cccagagccg tgatcgacta ttctaaggca gatgcatggg cagtgggagc catcgcctac
     2521 gagatcttcg gcctggtgaa tcccttctac ggacagggca aggcacacct ggagtcccgg
     2581 tcttatcagg aggcccagct gccagccctg ccagagtccg tgccacctga cgtgaggcag
     2641 ctggtgcgcg ccctgctgca gagagaggca agcaagaggc catccgccag agtggcagca
     2701 aacgtgctgc acctgagcct gtggggcgag cacatcctgg ccctgaagaa tctgaagctg
     2761 gataagatgg tgggatggct gctgcagcag tccgccgcca ccctgctggc aaacaggctg
     2821 acagagaagt gctgcgtgga gaccaagatg aagatgctgt ttctggccaa tctggagtgc
     2881 gagaccctgt gccaggccgc cctgctgctg tgcagctggc gcgccgccct ggtttctcaa
     2941 ggcagttcag gtggtggcgg gagcggaggt ggaggctcga gcggtgtgac cggctaccgg
     3001 ctgttcgagg agattctgta aggccgcgac tctagagtcg acctgcaggc atgcaagctg
     3061 atttggctgc tgccaccgct gagcaaggcc gcttcgagca gacatgataa gatacattga
     3121 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     3181 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
     3241 ttgcattcat tttatgtttc aggttcaggg ggagatgtgg gaggtttttt taagcaagta
     3301 aaacctctac aaatgtggta aaatcttaat taagaattct aatataactt cgtatagcat
     3361 acattatacg aagttatgga tccactgttg acaattaatc atcggcatag tatatcggca
     3421 tagtataata cgacaaggtg aggaactaaa cccaggaggc agatcatgat tgaacaagat
     3481 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     3541 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     3601 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     3661 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     3721 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     3781 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     3841 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     3901 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     3961 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     4021 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     4081 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     4141 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     4201 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     4261 ttcgaaatga ccgaccaagc gacgcccaac cggtatcagc tcactcaaag gcggtaatac
     4321 ggttatccac agaatcaggg gataacgcag gaaagaacat gtcaaacatg agaattacaa
     4381 cttatatcgt atggggctga cttcaggtgc tacatttgaa gagataaatt gcactgaaat
     4441 ctagaaatat tttatctgat taataagatg atcttcttga gatcgttttg gtctgcgcgt
     4501 aatctcttgc tctgaaaacg aaaaaaccgc cttgcagggc ggtttttcga aggttctctg
     4561 agctaccaac tctttgaacc gaggtaactg gcttggagga gcgcagtcac caaaacttgt
     4621 cctttcagtt tagccttaac cggcgcatga cttcaagact aactcctcta aatcaattac
     4681 cagtggctgc tgccagtggt gcttttgcat gtctttccgg gttggactca agacgatagt
     4741 taccggataa ggcgcagcgg tcggactgaa cggggggttc gtgcatacag tccagcttgg
     4801 agcgaactgc ctacccggaa ctgagtgtca ggcgtggaat gagacaaacg cggccataac
     4861 agcggaatga caccggtaaa ccgaaaggca ggaacaggag agcgcacgag ggagccgcca
     4921 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccaccactg atttgagcgt
     4981 cagatttcgt gatgcttgtc aggggggcgg agcctatgga aaaacggctt tgccgcggcc
     5041 ctctcacttc cctgttaagt atcttcctgg catcttccag gaaatctccg ccccgttcgt
     5101 aagccatttc cgctcgccgc agtcgaacga ccgagcgtag cgagtcagtg agcgaggaag
     5161 cggaatatat cctgtatcac atattctgct gacgcaccgg tgcagccttt tttctcctgc
     5221 cacatgaagc acttcactga caccctcatc agtgccaaca tagtaagcca gtatacactc
     5281 cgctagcgct gatgtccgga atcgtcacac aaaaaaccaa cacacagatg taatgaaaat
     5341 aaagatattt tattgcggcc atcgtgatgg ctagagttta gccctcccac acataaccag
     5401 agggcagcaa ttcacgaatc ccaactgccg tcggctgtcc atcactgtcc ttcactatgg
     5461 ctttgatccc aggatgcaga tcgagaagca cctgtcggca ccgtccgcag gggctcaaga
     5521 tgcccctgtt ctcatttccg atcgcgacga tacaagtcag gttgccagct gccgcagcag
     5581 cagcagtgcc cagcaccacg agttctgcac aaggtccccc agtaaaatga tatacattga
     5641 caccagtgaa gatgcggccg tcgctagaga gagctgcgct ggcgacgctg tagtcttcag
     5701 agatggggat gctgttgatt gtagccgttg ctctttcaat gagggtggat tcttcttgag
     5761 acaaaggctt ggccatggtg gcgctagtgt cagaagaatc gagctttttg caaaagccta
     5821 ggcctccaaa aaagcctcct cactacttct ggaatagctc agaggccgag gcggcctcgg
     5881 cctctgcata aataaaaaaa attagtcagc catggggcgg agaatgggcg gaactgggcg
     5941 gagttagggg cgggatgggc ggagttaggg gcgggactat ggttgctgac taattgagat
     6001 gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac ctggttgctg
     6061 actaattgag atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac
     6121 accctaactg acacacattc cacagctggt tctttccgcc tcagaaggta cctaaccaag
     6181 ttcctctttc agaggttatt tcaggccatg gtgctgcgc
//
