LOCUS       Exported                5459 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 5459)
  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..5459
                     /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..2409
                     /label=NCK1
                     /note="NCK1"
     misc_feature    1282..1461
                     /label=SH1
                     /note="SH1"
     misc_feature    2410..2417
                     /label=PmeI site
                     /note="Used in Flexi Vector Cloning"
     polyA_signal    2513..2734
                     /label=Sv40 late polyadenylation signal
     misc_feature    complement(2758..2791)
                     /label=Cre site
                     /note="Cre site"
     promoter        2800..2866
                     /label=EM7 bacterial promoter
     CDS             2880..3674
                     /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      3829..3865
                     /direction=RIGHT
                     /label=ColE1-derived plasmid replication origin
                     /note="The actual origin is the first base."
     polyA_signal    complement(4547..4594)
                     /label=Synthetic PolyAdenylation Signal
     misc_feature    complement(4618..5016)
                     /label=BlastR
                     /note="BlastR"
     misc_feature    complement(5041..5459)
                     /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 ggcagaagaa gtggtggtag tagccaaatt tgattatgtg
     1321 gcccaacaag aacaagagtt ggacatcaag aagaatgaga gattatggct tctggatgat
     1381 tctaagtcct ggtggcgagt tcgaaattcc atgaataaaa caggttttgt gccttctaac
     1441 tatgtggaaa ggaaaaacag tgctcggaaa gcatctattg tgaaaaacct aaaggatacc
     1501 ttaggcattg gaaaagtgaa aagaaaacct agtgtgccag attctgcatc tcctgctgat
     1561 gatagttttg ttgacccagg ggaacgtctc tatgacctca acatgcccgc ttatgtgaaa
     1621 tttaactaca tggctgagag agaggatgaa ttatcattga taaaggggac aaaggtgatc
     1681 gtcatggaga aatgcagtga tgggtggtgg cgtggtagct acaatggaca agttggatgg
     1741 ttcccttcaa actatgtaac tgaagaaggt gacagtcctt tgggtgacca tgtgggttct
     1801 ctgtcagaga aattagcagc agtcgtcaat aacctaaata ctgggcaagt gttgcatgtg
     1861 gtacaggctc tttacccatt cagctcatct aatgatgaag aacttaattt cgagaaagga
     1921 gatgtaatgg atgttattga aaaacctgaa aatgacccag agtggtggaa atgcaggaag
     1981 atcaatggta tggttggtct agtaccaaaa aactatgtta ccgttatgca gaataatcca
     2041 ttaacttcag gtttggaacc atcacctcca cagtgtgatt acattaggcc ttcactcact
     2101 ggaaagtttg ctggcaatcc ttggtattat ggcaaagtca ccaggcatca agcagaaatg
     2161 gcattaaatg aaagaggaca tgaaggggat ttcctcattc gtgatagtga atcttcgcca
     2221 aatgatttct cagtatcact aaaagcacaa gggaaaaaca agcattttaa agtccaacta
     2281 aaagagactg tctactgcat tgggcagcgt aaattcagca ccatggaaga acttgtagaa
     2341 cattacaaaa aggcaccaat ttttacaagt gaacaaggag aaaaattata tcttgtcaag
     2401 catttatcag tttaaacgaa ttcgggctcg gtacccgggg atcctctaga gtcgacctgc
     2461 aggcatgcaa gctgatttgg ctgctgccac cgctgagcaa ggccgcttcg agcagacatg
     2521 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
     2581 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
     2641 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggagat gtgggaggtt
     2701 tttttaagca agtaaaacct ctacaaatgt ggtaaaatct taattaagaa ttctaatata
     2761 acttcgtata gcatacatta tacgaagtta tggatccact gttgacaatt aatcatcggc
     2821 atagtatatc ggcatagtat aatacgacaa ggtgaggaac taaacccagg aggcagatca
     2881 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     2941 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     3001 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     3061 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     3121 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     3181 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     3241 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     3301 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     3361 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
     3421 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     3481 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     3541 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     3601 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     3661 acgagttctt ctgattcgaa atgaccgacc aagcgacgcc caaccggtat cagctcactc
     3721 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     3781 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     3841 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     3901 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     3961 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     4021 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     4081 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     4141 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     4201 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     4261 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     4321 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     4381 ttgcaagcag cagattacgc gcagaaaaaa aggatttcaa gaagatcctt tgatcttttc
     4441 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     4501 atcaaaaagg atcttcacct agatcctttt atagtccgga atcgtcacac aaaaaaccaa
     4561 cacacagatg taatgaaaat aaagatattt tattgcggcc atcgtgatgg ctagagttta
     4621 gccctcccac acataaccag agggcagcaa ttcacgaatc ccaactgccg tcggctgtcc
     4681 atcactgtcc ttcactatgg ctttgatccc aggatgcaga tcgagaagca cctgtcggca
     4741 ccgtccgcag gggctcaaga tgcccctgtt ctcatttccg atcgcgacga tacaagtcag
     4801 gttgccagct gccgcagcag cagcagtgcc cagcaccacg agttctgcac aaggtccccc
     4861 agtaaaatga tatacattga caccagtgaa gatgcggccg tcgctagaga gagctgcgct
     4921 ggcgacgctg tagtcttcag agatggggat gctgttgatt gtagccgttg ctctttcaat
     4981 gagggtggat tcttcttgag acaaaggctt ggccatggtg gcgctagtgt cagaagaatc
     5041 gagctttttg caaaagccta ggcctccaaa aaagcctcct cactacttct ggaatagctc
     5101 agaggccgag gcggcctcgg cctctgcata aataaaaaaa attagtcagc catggggcgg
     5161 agaatgggcg gaactgggcg gagttagggg cgggatgggc ggagttaggg gcgggactat
     5221 ggttgctgac taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga
     5281 ctttccacac ctggttgctg actaattgag atgcatgctt tgcatacttc tgcctgctgg
     5341 ggagcctggg gactttccac accctaactg acacacattc cacagctggt tctttccgcc
     5401 tcagaaggta cctaaccaag ttcctctttc agaggttatt tcaggccatg gtgctgcgc
//
