LOCUS       Exported                5714 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 5714)
  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..5714
                     /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..1729)
                     /label=VTA1
                     /note="VTA1"
     misc_feature    complement(1731..1738)
                     /label=SgfI Site
                     /note="Used in Flexi Vector Cloning"
     misc_feature    1743..1769
                     /label=B globin leader
                     /note="B globin leader"
     misc_feature    complement(1770..1786)
                     /label=SP6 promoter
                     /note="SP6 promoter"
     misc_feature    complement(1789..1834)
                     /label=Minimal CMV Promoter
     promoter        1844..2398
                     /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        2416..2482
                     /label=EM7 bacterial promoter
     CDS             2496..3290
                     /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      3445..3481
                     /direction=RIGHT
                     /label=ColE1-derived plasmid replication origin
                     /note="The actual origin is the first base."
     polyA_signal    complement(4163..4210)
                     /label=Synthetic PolyAdenylation Signal
     CDS             complement(4234..5271)
                     /label=HygroR
                     /note=" Feature found by back translation of amino acid 
                     feature definition."
     misc_feature    complement(5296..5714)
                     /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 agagaaactt ctctgcctgt cgtcagtaac ttgagagcct
      841 tttgtagatt ctggacagca gtgcttacat cttcatactg caaagcactg ccagcatatt
      901 tgcagtactt ctgagctcta gcaaagtctt ctggggttag acgaacatcc ccctgggaaa
      961 ttgtattgaa aagtgcggga tcaatggcag gtatagtctg tggagtaggt tggatagtat
     1021 tacttgctac acctgtgctg tgaggcactt ctgctggtgt attagctgga gcgtgtgcac
     1081 ccggaggaat ctgtattcca gtatagttgc ctgatggcat gttgcttggg tcataagttg
     1141 aagatgatga tggctgagtt ggctgagtgg gcagagaggc tgctccagca tcttcatttt
     1201 cttcaatatc attatcttct tcaattccaa cagggcctgc ttgaggagtc tccccattct
     1261 ttaaacaatt atggatgtat gttgccttcc atctggcata cttcctgtgt ttcacatttt
     1321 catcagtgag ttctccaaat actgttatga catctatcaa aagacttgca gtatagaagg
     1381 acttgatcat gtttttgtga aatcgtccag cacgatcttc attgtctgca tacaaaaaca
     1441 ttttcaaagc ataattctcc aaatgggcac agcccactat ttcttgagta atagcttcat
     1501 tatcacccaa ctgcttcttt agagcttcta actgatccat taactttgat aaaaatttgc
     1561 gacattcagg agttttacta tcgatcttca ttccagtctg cattgcgtat aaacgacagt
     1621 aataagccac cacagggtct cgcttgtcat gctcctgagc cgtcctcaga tgatgctgta
     1681 tgctcttgaa ctgtgcgggg agcgggggca gcggtgcaag cgcggccatg gcgatcgcta
     1741 gctgcaaaaa gaacaagtag cttgtattct atagtgtcac ctaaatgggc gatctgacgg
     1801 ttcactaaac cagctctgct tatataggcc tccctggcgc gccactagtt attaatagta
     1861 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
     1921 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
     1981 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
     2041 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagtc cgccccctat
     2101 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttacggga
     2161 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
     2221 ttggcagtac accaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
     2281 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
     2341 tcgtaataac cccgccccgt tgacgcaaat gggcggtagg cgtgtacggt gggaggtcaa
     2401 atcgaattct aatactgttg acaattaatc atcggcatag tatatcggca tagtataata
     2461 cgacaaggtg aggaactaaa cccaggaggc agatcatgat tgaacaagat ggattgcacg
     2521 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     2581 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     2641 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt
     2701 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     2761 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     2821 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     2881 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     2941 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     3001 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg
     3061 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     3121 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     3181 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     3241 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga ttcgaaatga
     3301 ccgaccaagc gacgcccaac cggtatcagc tcactcaaag gcggtaatac ggttatccac
     3361 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     3421 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     3481 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     3541 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     3601 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     3661 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     3721 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     3781 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     3841 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     3901 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     3961 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     4021 aaaaaaagga tttcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     4081 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     4141 ccttttatag tccggaatcg tcacacaaaa aaccaacaca cagatgtaat gaaaataaag
     4201 atattttatt gcggccatcg tgatggctag agtctaaact cgacctacct ccttagcgcg
     4261 cggtcgtgta ctgggccggc ggttgccgct gtcggccagc acctcgacgc agccgtcggt
     4321 ccatacggct gcgctccggc gagcgatttg tgtgcgaccg acggtgccgg ccccgctgcg
     4381 gacgatggca tcgcagcggc cttgagccca ggcagcatcg tcgaagttgc cgtccacgag
     4441 gctctggtag agctgatcca ggccgatgcg cagcatgtag gctcgcagac gagggctgcc
     4501 ggccagctcg ggatgccggc gctcgaagta gcgagtctgc tgctccatgc aagccagcca
     4561 gggccgccag aagaagatgt tggccacctc gtactgactg tccccgaaca tagcttcgga
     4621 ccagtcgatt acggcggtga tgcggccgtt gtcggtcagg acgttgttgc tgccgaagtc
     4681 ggcatggacg aggtggcgca cctcgggaca gtcttcggcc cacagcatca gttcgtccag
     4741 ggcttgagct acgctggcgg acacggtgtc gtccatcacg gtctgccagt ggtagacatg
     4801 gggatcagca atggcgcaaa tgaaatcccg ccaggttgtg tactggccga tgccttgggg
     4861 cccgaatggg ccgaagccgc tggtttgact gaggtcggcg gcggcaatag catccatagc
     4921 ttcggcgaca ggctgtaaca cagcaggcag ctctgtttcg gggaggtctt ggagagtaac
     4981 gccttgggcg cgtctactga tgcagtatgt caggctctcg ctgaactcgc cgatgtccaa
     5041 cacttcgggg atgggtagtg cagcgctggc gaagtggcgg tacacgtagc ggtctttgta
     5101 gaagccatca gcgcagctat tcacccgcag tacatagccg cgtccgccga catcgaagct
     5161 gaaggctcgg ctctcttcgc cctccgacaa ctgcatcagg tcgctcacac tgtcgaactt
     5221 ctcgatgaga aatttttcaa cgctggtagc ggtgagttcg ggcttcttca tggtggcgct
     5281 agtgtcagaa gaatcgagct ttttgcaaaa gcctaggcct ccaaaaaagc ctcctcacta
     5341 cttctggaat agctcagagg ccgaggcggc ctcggcctct gcataaataa aaaaaattag
     5401 tcagccatgg ggcggagaat gggcggaact gggcggagtt aggggcggga tgggcggagt
     5461 taggggcggg actatggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc
     5521 tggggagcct ggggactttc cacacctggt tgctgactaa ttgagatgca tgctttgcat
     5581 acttctgcct gctggggagc ctggggactt tccacaccct aactgacaca cattccacag
     5641 ctggttcttt ccgcctcaga aggtacctaa ccaagttcct ctttcagagg ttatttcagg
     5701 ccatggtgct gcgc
//
