LOCUS       Exported                4769 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 4769)
  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..4769
                     /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"
     gene            1279..1686
                     /label=Histone H3.3
                     /note="H3F3A"
     misc_feature    1687..1694
                     /label=PmeI site
                     /note="Used in Flexi Vector Cloning"
     polyA_signal    1823..2044
                     /label=Sv40 late polyadenylation signal
     promoter        2110..2176
                     /label=EM7 bacterial promoter
     CDS             2190..2984
                     /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      3139..3175
                     /direction=RIGHT
                     /label=ColE1-derived plasmid replication origin
                     /note="The actual origin is the first base."
     polyA_signal    complement(3857..3904)
                     /label=Synthetic PolyAdenylation Signal
     misc_feature    complement(3928..4326)
                     /label=BlastR
                     /note="BlastR"
     misc_feature    complement(4351..4769)
                     /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 ggctcgtaca aagcagactg cccgcaaatc gaccggtggt
     1321 aaagcaccca ggaagcaact ggctacaaaa gccgctcgca agagtgcgcc ctctactgga
     1381 ggggtgaaga aacctcatcg ttacaggcct ggtactgtgg cgctccgtga aattagacgt
     1441 tatcagaagt ccactgaact tctgattcgc aaacttccct tccagcgtct ggtgcgagaa
     1501 attgctcagg actttaaaac agatctgcgc ttccagagcg cagctatcgg tgctttgcag
     1561 gaggcaagtg aggcctatct ggttggcctt tttgaagaca ccaacctgtg tgctatccat
     1621 gccaaacgtg taacaattat gccaaaagac atccagctag cacgccgcat acgtggagaa
     1681 cgtgctgttt aaacgaattc gggctcggta cccggggatc ctctagagtc gacctgcagg
     1741 catgcaagct gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac
     1801 cgctgagcaa ggccgcttcg agcagacatg ataagataca ttgatgagtt tggacaaacc
     1861 acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta
     1921 tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg
     1981 tttcaggttc agggggagat gtgggaggtt tttttaagca agtaaaacct ctacaaatgt
     2041 ggtaaaatct taattaagaa ttctaatata acttcgtata gcatacatta tacgaagtta
     2101 tggatccact gttgacaatt aatcatcggc atagtatatc ggcatagtat aatacgacaa
     2161 ggtgaggaac taaacccagg aggcagatca tgattgaaca agatggattg cacgcaggtt
     2221 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     2281 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     2341 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
     2401 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     2461 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     2521 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     2581 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     2641 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     2701 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     2761 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     2821 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     2881 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     2941 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgattcgaa atgaccgacc
     3001 aagcgacgcc caaccggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     3061 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     3121 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     3181 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     3241 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     3301 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     3361 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     3421 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     3481 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     3541 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     3601 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     3661 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     3721 aggatttcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     3781 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     3841 atagtccgga atcgtcacac aaaaaaccaa cacacagatg taatgaaaat aaagatattt
     3901 tattgcggcc atcgtgatgg ctagagttta gccctcccac acataaccag agggcagcaa
     3961 ttcacgaatc ccaactgccg tcggctgtcc atcactgtcc ttcactatgg ctttgatccc
     4021 aggatgcaga tcgagaagca cctgtcggca ccgtccgcag gggctcaaga tgcccctgtt
     4081 ctcatttccg atcgcgacga tacaagtcag gttgccagct gccgcagcag cagcagtgcc
     4141 cagcaccacg agttctgcac aaggtccccc agtaaaatga tatacattga caccagtgaa
     4201 gatgcggccg tcgctagaga gagctgcgct ggcgacgctg tagtcttcag agatggggat
     4261 gctgttgatt gtagccgttg ctctttcaat gagggtggat tcttcttgag acaaaggctt
     4321 ggccatggtg gcgctagtgt cagaagaatc gagctttttg caaaagccta ggcctccaaa
     4381 aaagcctcct cactacttct ggaatagctc agaggccgag gcggcctcgg cctctgcata
     4441 aataaaaaaa attagtcagc catggggcgg agaatgggcg gaactgggcg gagttagggg
     4501 cgggatgggc ggagttaggg gcgggactat ggttgctgac taattgagat gcatgctttg
     4561 catacttctg cctgctgggg agcctgggga ctttccacac ctggttgctg actaattgag
     4621 atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac accctaactg
     4681 acacacattc cacagctggt tctttccgcc tcagaaggta cctaaccaag ttcctctttc
     4741 agaggttatt tcaggccatg gtgctgcgc
//
