LOCUS       Exported                5501 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 5501)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Friday, May 30, 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..5501
                     /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..985)
                     /label=BCL2L2
                     /note="BCL2L2"
     misc_feature    complement(987..994)
                     /label=SgfI Site
                     /note="Used in Flexi Vector Cloning"
     CDS             complement(995..1039)
                     /codon_start=1
                     /label=15GS linker
                     /note="15GS linker"
                     /translation="GSSGGGGSGGGGSSG"
     gene            complement(1040..1516)
                     /label=LgBiT
                     /note="LgBiT"
     misc_feature    1530..1556
                     /label=B globin leader
                     /note="B globin leader"
     misc_feature    complement(1557..1573)
                     /label=SP6 promoter
                     /note="SP6 promoter"
     misc_feature    complement(1576..1621)
                     /label=Minimal CMV Promoter 2
     misc_feature    1599..1621
                     /label=core
                     /note="core"
     promoter        1631..2185
                     /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."
     promoter        2203..2269
                     /label=EM7 bacterial promoter
     misc_feature    2283..3077
                     /label=KanR
                     /note="KanR"
     rep_origin      3232..3268
                     /direction=RIGHT
                     /label=ColE1-derived plasmid replication origin
                     /note="The actual origin is the first base."
     polyA_signal    complement(3950..3997)
                     /label=Synthetic PolyAdenylation Signal
     CDS             complement(4021..5058)
                     /label=HygroR
                     /note=" Feature found by back translation of amino acid 
                     feature definition."
     misc_feature    complement(5083..5501)
                     /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 ttaaaccttg ctagcaaaaa
      421 aggcccctac agttaccagg gcccccagtg ccacggcccc cgtcagaact gtcctcactg
      481 atgcccagtt accctcccgc agacgccgcg cctcttccag agccccgtca ccgtatagag
      541 ctgtgaactc cgcccagccc ccactgctgt ggatccagtc agccagccgc gtctccaggt
      601 aggccaccat ccactcctgc acttgtccca ccagtggttc catctccttg ttgacactct
      661 cagcacacag tgcagcccca aagacaaaga aggctacaag gcggccccag ttggggcccc
      721 cttgaaaaag ttcatcggag acctgggtga agcgttgttg ggctgagcct ggggtcacat
      781 gcagctgagc cgccagatca gagaaggtgc gccggaagcg ggtctcgaac tcatctccag
      841 ctgcccgcat ggcttggtgc agcgggtcag ctgctgggcc ctccccgggg ccagctccac
      901 agacataacc cttctgcctc agcttataac ctacaaagtc tgccaccaga gcccgtgtgt
      961 ctggggccga ggctggggtc gccatggcga tcgcaccgct cgagcctcca cctccgctcc
     1021 cgccaccacc ggagctccca ctgttgatgg ttactcggaa cagcatggag ccgtcggggg
     1081 tgatcaggcg ctcgtcgata attttgttgc cgttccacag ggtccctgtt acagtgatct
     1141 ttttgccgtc gaacacggcg atgccttcat acggccgtcc gaaatagttc agcatgttcg
     1201 gcgtaacccc gtcgattacc agtgtgccat agggcaggat caccttaaag tgatgatcat
     1261 ccacagggta caccacctta aacacctctt cgatctgggc catttggtcg gcgctcagac
     1321 cttcatacgg gatgatgaca tggatgtcga tcttcagggc attttcaccg ctccggacaa
     1381 tcctttggat cggagttacg gacacggcga gattctgcag caaactggac acacctccct
     1441 gttcaaggac ttggtccagg ttgtaggcgg ctgtctgttc ccagtcccca acgaaatctt
     1501 cgagtgtgaa gaccatggtg agcgctagct gcaaaaagaa caagtagctt gtattctata
     1561 gtgtcaccta aatgggcgat ctgacggttc actaaaccag ctctgcttat ataggcctcc
     1621 ctggcgcgcc actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
     1681 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
     1741 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
     1801 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
     1861 gtatcatatg ccaagtccgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     1921 ttatgcccag tacatgacct tacgggactt tcctacttgg cagtacatct acgtattagt
     1981 catcgctatt accatggtga tgcggttttg gcagtacacc aatgggcgtg gatagcggtt
     2041 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
     2101 ccaaaatcaa cgggactttc caaaatgtcg taataacccc gccccgttga cgcaaatggg
     2161 cggtaggcgt gtacggtggg aggtcaaatc gaattctaat actgttgaca attaatcatc
     2221 ggcatagtat atcggcatag tataatacga caaggtgagg aactaaaccc aggaggcaga
     2281 tcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     2341 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     2401 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     2461 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     2521 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     2581 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     2641 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     2701 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     2761 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
     2821 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     2881 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     2941 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     3001 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     3061 ttgacgagtt cttctgattc gaaatgaccg accaagcgac gcccaaccgg tatcagctca
     3121 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     3181 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     3241 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     3301 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     3361 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     3421 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     3481 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     3541 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     3601 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     3661 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     3721 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     3781 tgtttgcaag cagcagatta cgcgcagaaa aaaaggattt caagaagatc ctttgatctt
     3841 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     3901 attatcaaaa aggatcttca cctagatcct tttatagtcc ggaatcgtca cacaaaaaac
     3961 caacacacag atgtaatgaa aataaagata ttttattgcg gccatcgtga tggctagagt
     4021 ctaaactcga cctacctcct tagcgcgcgg tcgtgtactg ggccggcggt tgccgctgtc
     4081 ggccagcacc tcgacgcagc cgtcggtcca tacggctgcg ctccggcgag cgatttgtgt
     4141 gcgaccgacg gtgccggccc cgctgcggac gatggcatcg cagcggcctt gagcccaggc
     4201 agcatcgtcg aagttgccgt ccacgaggct ctggtagagc tgatccaggc cgatgcgcag
     4261 catgtaggct cgcagacgag ggctgccggc cagctcggga tgccggcgct cgaagtagcg
     4321 agtctgctgc tccatgcaag ccagccaggg ccgccagaag aagatgttgg ccacctcgta
     4381 ctgactgtcc ccgaacatag cttcggacca gtcgattacg gcggtgatgc ggccgttgtc
     4441 ggtcaggacg ttgttgctgc cgaagtcggc atggacgagg tggcgcacct cgggacagtc
     4501 ttcggcccac agcatcagtt cgtccagggc ttgagctacg ctggcggaca cggtgtcgtc
     4561 catcacggtc tgccagtggt agacatgggg atcagcaatg gcgcaaatga aatcccgcca
     4621 ggttgtgtac tggccgatgc cttggggccc gaatgggccg aagccgctgg tttgactgag
     4681 gtcggcggcg gcaatagcat ccatagcttc ggcgacaggc tgtaacacag caggcagctc
     4741 tgtttcgggg aggtcttgga gagtaacgcc ttgggcgcgt ctactgatgc agtatgtcag
     4801 gctctcgctg aactcgccga tgtccaacac ttcggggatg ggtagtgcag cgctggcgaa
     4861 gtggcggtac acgtagcggt ctttgtagaa gccatcagcg cagctattca cccgcagtac
     4921 atagccgcgt ccgccgacat cgaagctgaa ggctcggctc tcttcgccct ccgacaactg
     4981 catcaggtcg ctcacactgt cgaacttctc gatgagaaat ttttcaacgc tggtagcggt
     5041 gagttcgggc ttcttcatgg tggcgctagt gtcagaagaa tcgagctttt tgcaaaagcc
     5101 taggcctcca aaaaagcctc ctcactactt ctggaatagc tcagaggccg aggcggcctc
     5161 ggcctctgca taaataaaaa aaattagtca gccatggggc ggagaatggg cggaactggg
     5221 cggagttagg ggcgggatgg gcggagttag gggcgggact atggttgctg actaattgag
     5281 atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac acctggttgc
     5341 tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc
     5401 acaccctaac tgacacacat tccacagctg gttctttccg cctcagaagg tacctaacca
     5461 agttcctctt tcagaggtta tttcaggcca tggtgctgcg c
//
