LOCUS       Exported                6284 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 6284)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Apr 15, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
COMMENT     pRSG193 designed by Brock F. Binkowski

            

            PCR amplify pF-BiBiTLn w/ BFBRSG11/BFBRSG4 (PCR #1; 2.0 kbp);

            PCR amplify pF-BiBiTsmC w/ BFBRSG5/BFBRSG9 (PCR #2; 2.8 kbp); 

            PCR amplify pF-4B w/ BFBRSG10/BFBRSG12 (464 bp);

            3X component Gibson Assembly;

            electroporation of BR610.
FEATURES             Location/Qualifiers
     source          1..6284
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    complement(15..202)
                     /label=SV40\polyA\signal
     misc_feature    complement(206..255)
                     /label=MCS 2
     misc_feature    complement(399..406)
                     /label=PmeI site
                     /note="Used in Flexi Vector Cloning"
     misc_feature    complement(407..1594)
                     /label=SLC35A2
                     /note="SLC35A2"
     misc_feature    complement(1596..1603)
                     /label=SgfI Site
                     /note="Used in Flexi Vector Cloning"
     CDS             complement(1604..1648)
                     /codon_start=1
                     /label=15GS linker
                     /note="15GS linker"
                     /translation="GSSGGGGSGGGGSSG"
     gene            complement(1649..2125)
                     /label=LgBiT
                     /note="LgBiT"
     misc_feature    2139..2165
                     /label=B globin leader
                     /note="B globin leader"
     misc_feature    complement(2166..2182)
                     /label=SP6 promoter
                     /note="SP6 promoter"
     misc_feature    complement(2185..2230)
                     /label=Minimal CMV Promoter
     misc_feature    2208..2230
                     /label=core
                     /note="core"
     promoter        2240..2794
                     /label=CMV Enhancer
                     /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."
     promoter        2812..2878
                     /label=EM7 bacterial promoter
     misc_feature    2892..3686
                     /label=KanR
                     /note="KanR"
     misc_feature    3789..4711
                     /label=P15 ori
                     /note="P15 ori"
     misc_feature    3966
                     /label=C2129
                     /note="C2129"
     polyA_signal    complement(4733..4780)
                     /label=Synthetic PolyAdenylation Signal
     CDS             complement(4804..5841)
                     /label=HygroR
                     /note=" Feature found by back translation of amino acid 
                     feature definition."
     misc_feature    complement(5866..6284)
                     /label=SV40 promoter
                     /note="SV40 promoter"
ORIGIN
        1 aagacgcgtg acgtcggcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
       61 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
      121 ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca
      181 aatgtggtat ggctgattat gatccgcggc cgcaatacgt cgacgttatc agctgacttc
      241 gtacgagagc ctaggtgttt ccccaagggg ttatgctagt tattgctcag cggtggcagc
      301 agccaactca gcttcctttc gggctttgtt agcagccgga tcagcttgca tgcctgcagg
      361 tcgactctag aggatccccg ggtaccgagc ccgaattcgt ttaaacggat cccttcactt
      421 tggtcagcaa tttcggcagg aagggctcgg ttatgaggtc accgcggtgg gaggacaact
      481 gtggaggcgg cggctgtccc ggaggttgct gatggacgca cggaccggag gcagacgcgg
      541 aagcagaggc tatggctttc gctgctcccc gaggcaggga gtacaaatag accgctccta
      601 tgacaagccc agcacccaag gcaaagagcg gatctacatg aaacccgaag agtcttattg
      661 aggcgacggt ggacaatacg atggacaagg aggtcgcgaa tcccttcagt atgttgtccg
      721 cgtatttcac aacaaccgcc acgagcaatc cgccaaatgc ttggttcaat acgacacccc
      781 aaacagctgg tgtgtatcca aaaaagaatc cgcgagttgc gactgcggtg ccctccgccc
      841 accacaaacc caccaacccg agggccgtcc caaagaggcc aagctgcaga tttctcaacc
      901 agacactacc agaagaaccc ttcaggatct tctcaaaata tacacccgcg aacccagagc
      961 taaggcagga ggccaccaca gccgcgagcc cggctcccgg attctgatcg agagggcggg
     1021 gacctcctcc tcccgcttgt tgagcttgaa ctatggcgac ccctgtaaag aggagaagga
     1081 gactggccca ttggagtcgg gacaaggacc ggttcagcat aagaacggag aacaatgccg
     1141 ttgtcaggat tttcagttgg taggtgactt ggaacgtagc tgccggcaaa tttgatatag
     1201 cgacatattg aagattattt tgcaaagtat aaatgaggct gggtaccgcc aacttcagtg
     1261 tgtcaacata ttggacaagc actgcctcgt gcagaaacag aaccaggtgt tttacatttc
     1321 ctcgtttttg cgcaaagaga aggagcaggc acgtcaatcc cttaaggacc tctgccatga
     1381 cgactgcggt tgttgcgaaa aatcgatcgc ccggcaacgt ccgagcgtat cttatagaaa
     1441 ggatgaggga agcattttgc actacgagaa cagcaagaga aatatacttc aatcttcggt
     1501 gggcggctga agcagtccca ggttcaagag cgcctgcaga caccgcgccc ggaccaggtg
     1561 ctgccgtact tcctcctgcg cccacggcag ccatggcgat cgcaccgctc gagcctccac
     1621 ctccgctccc gccaccaccg gagctcccac tgttgatggt tactcggaac agcatggagc
     1681 cgtcgggggt gatcaggcgc tcgtcgataa ttttgttgcc gttccacagg gtccctgtta
     1741 cagtgatctt tttgccgtcg aacacggcga tgccttcata cggccgtccg aaatagttca
     1801 gcatgttcgg cgtaaccccg tcgattacca gtgtgccata gggcaggatc accttaaagt
     1861 gatgatcatc cacagggtac accaccttaa acacctcttc gatctgggcc atttggtcgg
     1921 cgctcagacc ttcatacggg atgatgacat ggatgtcgat cttcagggca ttttcaccgc
     1981 tccggacaat cctttggatc ggagttacgg acacggcgag attctgcagc aaactggaca
     2041 cacctccctg ttcaaggact tggtccaggt tgtaggcggc tgtctgttcc cagtccccaa
     2101 cgaaatcttc gagtgtgaag accatggtga gcgctagctg caaaaagaac aagtagcttg
     2161 tattctatag tgtcacctaa atgggcgatc tgacggttca ctaaaccagc tctgcttata
     2221 taggcctccc tggcgcgcca ctagttatta atagtaatca attacggggt cattagttca
     2281 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
     2341 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
     2401 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
     2461 acatcaagtg tatcatatgc caagtccgcc ccctattgac gtcaatgacg gtaaatggcc
     2521 cgcctggcat tatgcccagt acatgacctt acgggacttt cctacttggc agtacatcta
     2581 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacacca atgggcgtgg
     2641 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
     2701 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aataaccccg ccccgttgac
     2761 gcaaatgggc ggtaggcgtg tacggtggga ggtcaaatcg aattctaata ctgttgacaa
     2821 ttaatcatcg gcatagtata tcggcatagt ataatacgac aaggtgagga actaaaccca
     2881 ggaggcagat catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     2941 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     3001 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     3061 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     3121 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     3181 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     3241 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     3301 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     3361 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     3421 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     3481 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     3541 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     3601 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     3661 atcgccttct tgacgagttc ttctgattcg aaatgaccga ccaagcgacg cccaaccggt
     3721 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     3781 gaacatgtca aacatgagaa ttacaactta tatcgtatgg ggctgacttc aggtgctaca
     3841 tttgaagaga taaattgcac tgaaatctag aaatatttta tctgattaat aagatgatct
     3901 tcttgagatc gttttggtct gcgcgtaatc tcttgctctg aaaacgaaaa aaccgccttg
     3961 cagggcggtt tttcgaaggt tctctgagct accaactctt tgaaccgagg taactggctt
     4021 ggaggagcgc agtcaccaaa acttgtcctt tcagtttagc cttaaccggc gcatgacttc
     4081 aagactaact cctctaaatc aattaccagt ggctgctgcc agtggtgctt ttgcatgtct
     4141 ttccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg actgaacggg
     4201 gggttcgtgc atacagtcca gcttggagcg aactgcctac ccggaactga gtgtcaggcg
     4261 tggaatgaga caaacgcggc cataacagcg gaatgacacc ggtaaaccga aaggcaggaa
     4321 caggagagcg cacgagggag ccgccagggg gaaacgcctg gtatctttat agtcctgtcg
     4381 ggtttcgcca ccactgattt gagcgtcaga tttcgtgatg cttgtcaggg gggcggagcc
     4441 tatggaaaaa cggctttgcc gcggccctct cacttccctg ttaagtatct tcctggcatc
     4501 ttccaggaaa tctccgcccc gttcgtaagc catttccgct cgccgcagtc gaacgaccga
     4561 gcgtagcgag tcagtgagcg aggaagcgga atatatcctg tatcacatat tctgctgacg
     4621 caccggtgca gccttttttc tcctgccaca tgaagcactt cactgacacc ctcatcagtg
     4681 ccaacatagt aagccagtat acactccgct agcgctgatg tccggaatcg tcacacaaaa
     4741 aaccaacaca cagatgtaat gaaaataaag atattttatt gcggccatcg tgatggctag
     4801 agtctaaact cgacctacct ccttagcgcg cggtcgtgta ctgggccggc ggttgccgct
     4861 gtcggccagc acctcgacgc agccgtcggt ccatacggct gcgctccggc gagcgatttg
     4921 tgtgcgaccg acggtgccgg ccccgctgcg gacgatggca tcgcagcggc cttgagccca
     4981 ggcagcatcg tcgaagttgc cgtccacgag gctctggtag agctgatcca ggccgatgcg
     5041 cagcatgtag gctcgcagac gagggctgcc ggccagctcg ggatgccggc gctcgaagta
     5101 gcgagtctgc tgctccatgc aagccagcca gggccgccag aagaagatgt tggccacctc
     5161 gtactgactg tccccgaaca tagcttcgga ccagtcgatt acggcggtga tgcggccgtt
     5221 gtcggtcagg acgttgttgc tgccgaagtc ggcatggacg aggtggcgca cctcgggaca
     5281 gtcttcggcc cacagcatca gttcgtccag ggcttgagct acgctggcgg acacggtgtc
     5341 gtccatcacg gtctgccagt ggtagacatg gggatcagca atggcgcaaa tgaaatcccg
     5401 ccaggttgtg tactggccga tgccttgggg cccgaatggg ccgaagccgc tggtttgact
     5461 gaggtcggcg gcggcaatag catccatagc ttcggcgaca ggctgtaaca cagcaggcag
     5521 ctctgtttcg gggaggtctt ggagagtaac gccttgggcg cgtctactga tgcagtatgt
     5581 caggctctcg ctgaactcgc cgatgtccaa cacttcgggg atgggtagtg cagcgctggc
     5641 gaagtggcgg tacacgtagc ggtctttgta gaagccatca gcgcagctat tcacccgcag
     5701 tacatagccg cgtccgccga catcgaagct gaaggctcgg ctctcttcgc cctccgacaa
     5761 ctgcatcagg tcgctcacac tgtcgaactt ctcgatgaga aatttttcaa cgctggtagc
     5821 ggtgagttcg ggcttcttca tggtggcgct agtgtcagaa gaatcgagct ttttgcaaaa
     5881 gcctaggcct ccaaaaaagc ctcctcacta cttctggaat agctcagagg ccgaggcggc
     5941 ctcggcctct gcataaataa aaaaaattag tcagccatgg ggcggagaat gggcggaact
     6001 gggcggagtt aggggcggga tgggcggagt taggggcggg actatggttg ctgactaatt
     6061 gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc cacacctggt
     6121 tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt
     6181 tccacaccct aactgacaca cattccacag ctggttcttt ccgcctcaga aggtacctaa
     6241 ccaagttcct ctttcagagg ttatttcagg ccatggtgct gcgc
//
