LOCUS       Exported                5465 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 5465)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Apr 15, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
COMMENT     pRSG192 designed by Brock F. Binkowski

            

            PCR amplify pF-BiBiTLc 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..5465
                     /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(278..787)
                     /label=NanoLuc
                     /note="NanoLuc"
     misc_feature    complement(788..802)
                     /label=linker
                     /note="linker"
     misc_feature    complement(809..1480)
                     /label=CHMP4B
                     /note="CHMP4B"
     misc_feature    complement(1482..1489)
                     /label=SgfI Site
                     /note="Used in Flexi Vector Cloning"
     misc_feature    1494..1520
                     /label=B globin leader
                     /note="B globin leader"
     misc_feature    complement(1521..1537)
                     /label=SP6 promoter
                     /note="SP6 promoter"
     misc_feature    complement(1540..1585)
                     /label=Minimal CMV Promoter
     promoter        1595..2149
                     /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        2167..2233
                     /label=EM7 bacterial promoter
     CDS             2247..3041
                     /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      3196..3232
                     /direction=RIGHT
                     /label=ColE1-derived plasmid replication origin
                     /note="The actual origin is the first base."
     polyA_signal    complement(3914..3961)
                     /label=Synthetic PolyAdenylation Signal
     CDS             complement(3985..5022)
                     /label=HygroR
                     /note=" Feature found by back translation of amino acid 
                     feature definition."
     misc_feature    complement(5047..5465)
                     /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 ctagagtcgc ggccttacgc cagaatgcgt tcgcacagcc
      301 gccagccggt cactccgttg atggttactc ggaacagcag ggagccgtcg gggttgatca
      361 ggcgctcgtc gataattttg ttgccgttcc acagggtccc tgttacagtg atctttttgc
      421 cgtcgaacac ggcgatgcct tcatacggcc gtccgaaata gtcgatcatg ttcggcgtaa
      481 ccccgtcgat taccagtgtg ccatagtgca ggatcacctt aaagtgatga tcatccacag
      541 ggtacaccac cttaaaaatt ttttcgatct ggcccatttg gtcgccgctc agaccttcat
      601 acgggatgat gacatggatg tcgatcttca gcccattttc accgctcagg acaatccttt
      661 ggatcggagt tacggacacc ccgagattct gaaacaaact ggacacacct ccctgttcaa
      721 ggacttggtc caggttgtag ccggctgtct gtcgccagtc cccaacgaaa tcttcgagtg
      781 tgaagacgcc gctcgagccg agagaaacca tggatccagc ccagttctcc aattccttca
      841 tgtcgtcgtc ctcctcttct ttcttcttgg cgggttttga tggtagggct atagagggaa
      901 catttggtag agggactgtt tcgggtccac tgatttccag caaattcttg tctagttcct
      961 cctgttctag ttcttctaat tccgccatga gctcatcctc gtcaaactct tctccaaacc
     1021 ctacaggttt cgaaattgct gttgaaatct cctctgcaag ttcttgctgg tcagcaatgt
     1081 cctgcattaa ctcatcaact ttatcgatgt ccatgttgtc atgggccgcc ttcatggcct
     1141 tggcggcata gcccatgttc ttgagcacct cggtgttggt gttggcattc tccagggcct
     1201 cccgctggaa ctcgatggtt gataatgtgc cgtcgatctg cgccagctgc ttctcatacc
     1261 tcttcttacg cttcagtgcc tggagggccg cgcgcttgtt tttggtgccg tgcttcttgg
     1321 cggccgtcag ctcctgctcg attttcttct ccaggaactc ctgtttcttg cttaacatct
     1381 cttccgtgtc ccgcagccgc tggatggcct cctggggggt cgggccgccc ttgccggcct
     1441 taccccctcc agccccgaac agcttcccga acaccgacat ggcgatcgct agctgcaaaa
     1501 agaacaagta gcttgtattc tatagtgtca cctaaatggg cgatctgacg gttcactaaa
     1561 ccagctctgc ttatataggc ctccctggcg cgccactagt tattaatagt aatcaattac
     1621 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
     1681 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
     1741 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
     1801 tgcccacttg gcagtacatc aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa
     1861 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttacggg actttcctac
     1921 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta
     1981 caccaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
     2041 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa
     2101 ccccgccccg ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtca aatcgaattc
     2161 taatactgtt gacaattaat catcggcata gtatatcggc atagtataat acgacaaggt
     2221 gaggaactaa acccaggagg cagatcatga ttgaacaaga tggattgcac gcaggttctc
     2281 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     2341 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     2401 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca
     2461 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     2521 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     2581 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     2641 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     2701 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     2761 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     2821 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     2881 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     2941 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     3001 agcgcatcgc cttctatcgc cttcttgacg agttcttctg attcgaaatg accgaccaag
     3061 cgacgcccaa ccggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     3121 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     3181 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     3241 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     3301 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     3361 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     3421 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     3481 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     3541 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     3601 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     3661 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     3721 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     3781 atttcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     3841 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttata
     3901 gtccggaatc gtcacacaaa aaaccaacac acagatgtaa tgaaaataaa gatattttat
     3961 tgcggccatc gtgatggcta gagtctaaac tcgacctacc tccttagcgc gcggtcgtgt
     4021 actgggccgg cggttgccgc tgtcggccag cacctcgacg cagccgtcgg tccatacggc
     4081 tgcgctccgg cgagcgattt gtgtgcgacc gacggtgccg gccccgctgc ggacgatggc
     4141 atcgcagcgg ccttgagccc aggcagcatc gtcgaagttg ccgtccacga ggctctggta
     4201 gagctgatcc aggccgatgc gcagcatgta ggctcgcaga cgagggctgc cggccagctc
     4261 gggatgccgg cgctcgaagt agcgagtctg ctgctccatg caagccagcc agggccgcca
     4321 gaagaagatg ttggccacct cgtactgact gtccccgaac atagcttcgg accagtcgat
     4381 tacggcggtg atgcggccgt tgtcggtcag gacgttgttg ctgccgaagt cggcatggac
     4441 gaggtggcgc acctcgggac agtcttcggc ccacagcatc agttcgtcca gggcttgagc
     4501 tacgctggcg gacacggtgt cgtccatcac ggtctgccag tggtagacat ggggatcagc
     4561 aatggcgcaa atgaaatccc gccaggttgt gtactggccg atgccttggg gcccgaatgg
     4621 gccgaagccg ctggtttgac tgaggtcggc ggcggcaata gcatccatag cttcggcgac
     4681 aggctgtaac acagcaggca gctctgtttc ggggaggtct tggagagtaa cgccttgggc
     4741 gcgtctactg atgcagtatg tcaggctctc gctgaactcg ccgatgtcca acacttcggg
     4801 gatgggtagt gcagcgctgg cgaagtggcg gtacacgtag cggtctttgt agaagccatc
     4861 agcgcagcta ttcacccgca gtacatagcc gcgtccgccg acatcgaagc tgaaggctcg
     4921 gctctcttcg ccctccgaca actgcatcag gtcgctcaca ctgtcgaact tctcgatgag
     4981 aaatttttca acgctggtag cggtgagttc gggcttcttc atggtggcgc tagtgtcaga
     5041 agaatcgagc tttttgcaaa agcctaggcc tccaaaaaag cctcctcact acttctggaa
     5101 tagctcagag gccgaggcgg cctcggcctc tgcataaata aaaaaaatta gtcagccatg
     5161 gggcggagaa tgggcggaac tgggcggagt taggggcggg atgggcggag ttaggggcgg
     5221 gactatggtt gctgactaat tgagatgcat gctttgcata cttctgcctg ctggggagcc
     5281 tggggacttt ccacacctgg ttgctgacta attgagatgc atgctttgca tacttctgcc
     5341 tgctggggag cctggggact ttccacaccc taactgacac acattccaca gctggttctt
     5401 tccgcctcag aaggtaccta accaagttcc tctttcagag gttatttcag gccatggtgc
     5461 tgcgc
//
