LOCUS       Exported                5303 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 5303)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Friday, May 30, 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..5303
                     /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
     gene            complement(278..751)
                     /label=LgBiT
                     /note="LgBiT"
     CDS             complement(752..796)
                     /codon_start=1
                     /label=15GS linker
                     /note="15GS linker"
                     /translation="GSSGGGGSGGGGSSG"
     misc_feature    complement(806..1144)
                     /label=DAP12 insert
                     /note="Recieved from Kazusa in August 2010. Entered into 
                     database semi automatically by Ethan in August 2010. 
                     Accession number NM_003332 added 11/4/2010 from BLAST and 
                     length similarity. (note added in Automated Blaster by 
                     11/4/2010 3:28:40 PM"
     misc_feature    complement(1146..1153)
                     /label=SgfI Site
                     /note="Used in Flexi Vector Cloning"
     misc_feature    1158..1184
                     /label=B globin leader
                     /note="B globin leader"
     misc_feature    complement(1185..1201)
                     /label=SP6 promoter
                     /note="SP6 promoter"
     misc_feature    complement(1204..1249)
                     /label=Minimal CMV Promoter
     misc_feature    1227..1249
                     /label=core
                     /note="core"
     promoter        1259..1813
                     /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        1831..1897
                     /label=EM7 bacterial promoter
     CDS             1911..2705
                     /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."
     misc_feature    2808..3730
                     /label=P15 ori
                     /note="P15 ori"
     misc_feature    2985
                     /label=C2129
                     /note="C2129"
     polyA_signal    complement(3752..3799)
                     /label=Synthetic PolyAdenylation Signal
     CDS             complement(3823..4860)
                     /label=HygroR
                     /note=" Feature found by back translation of amino acid 
                     feature definition."
     misc_feature    complement(4885..5303)
                     /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 ggccttagct gttgatggtt actcggaaca
      301 gcatggagcc gtcgggggtg atcaggcgct cgtcgataat tttgttgccg ttccacaggg
      361 tccctgttac agtgatcttt ttgccgtcga acacggcgat gccttcatac ggccgtccga
      421 aatagttcag catgttcggc gtaaccccgt cgattaccag tgtgccatag ggcaggatca
      481 ccttaaagtg atgatcatcc acagggtaca ccaccttaaa cacctcttcg atctgggcca
      541 tttggtcggc gctcagacct tcatacggga tgatgacatg gatgtcgatc ttcagggcat
      601 tttcaccgct ccggacaatc ctttggatcg gagttacgga cacggcgaga ttctgcagca
      661 aactggacac acctccctgt tcaaggactt ggtccaggtt gtaggcggct gtctgttccc
      721 agtccccaac gaaatcttcg agtgtgaaga caccgctcga gcctccacct ccgctcccgc
      781 caccacctga actgccttga gaaactttgt aatacggcct ctgtgtgttg aggtcgctgt
      841 agacatccga cctctgaccc tggagctcct gataaggcga ctcggtctca gtgatacgct
      901 gtttccgggt cgctgcctcc gcagcccctc gcccccgagg gaccagccgg cccaggaagt
      961 acacggccag ggcaatgagc actgtcagca ccaggtctcc catcacgatc cctgccagca
     1021 cgcccgggct caccgtagag caactgcaat cgctctgggc ctgggcctgg acaggacgga
     1081 gaccacttac agccagcagg agaggcagga gcaggagcct gctgcagggt tcaagtcccc
     1141 ccatggcgat cgctagctgc aaaaagaaca agtagcttgt attctatagt gtcacctaaa
     1201 tgggcgatct gacggttcac taaaccagct ctgcttatat aggcctccct ggcgcgccac
     1261 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
     1321 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
     1381 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
     1441 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
     1501 aagtccgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
     1561 catgacctta cgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
     1621 catggtgatg cggttttggc agtacaccaa tgggcgtgga tagcggtttg actcacgggg
     1681 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
     1741 ggactttcca aaatgtcgta ataaccccgc cccgttgacg caaatgggcg gtaggcgtgt
     1801 acggtgggag gtcaaatcga attctaatac tgttgacaat taatcatcgg catagtatat
     1861 cggcatagta taatacgaca aggtgaggaa ctaaacccag gaggcagatc atgattgaac
     1921 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     1981 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     2041 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
     2101 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     2161 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     2221 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     2281 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     2341 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     2401 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc
     2461 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     2521 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     2581 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     2641 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     2701 tctgattcga aatgaccgac caagcgacgc ccaaccggta tcagctcact caaaggcggt
     2761 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtcaa acatgagaat
     2821 tacaacttat atcgtatggg gctgacttca ggtgctacat ttgaagagat aaattgcact
     2881 gaaatctaga aatattttat ctgattaata agatgatctt cttgagatcg ttttggtctg
     2941 cgcgtaatct cttgctctga aaacgaaaaa accgccttgc agggcggttt ttcgaaggtt
     3001 ctctgagcta ccaactcttt gaaccgaggt aactggcttg gaggagcgca gtcaccaaaa
     3061 cttgtccttt cagtttagcc ttaaccggcg catgacttca agactaactc ctctaaatca
     3121 attaccagtg gctgctgcca gtggtgcttt tgcatgtctt tccgggttgg actcaagacg
     3181 atagttaccg gataaggcgc agcggtcgga ctgaacgggg ggttcgtgca tacagtccag
     3241 cttggagcga actgcctacc cggaactgag tgtcaggcgt ggaatgagac aaacgcggcc
     3301 ataacagcgg aatgacaccg gtaaaccgaa aggcaggaac aggagagcgc acgagggagc
     3361 cgccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac cactgatttg
     3421 agcgtcagat ttcgtgatgc ttgtcagggg ggcggagcct atggaaaaac ggctttgccg
     3481 cggccctctc acttccctgt taagtatctt cctggcatct tccaggaaat ctccgccccg
     3541 ttcgtaagcc atttccgctc gccgcagtcg aacgaccgag cgtagcgagt cagtgagcga
     3601 ggaagcggaa tatatcctgt atcacatatt ctgctgacgc accggtgcag ccttttttct
     3661 cctgccacat gaagcacttc actgacaccc tcatcagtgc caacatagta agccagtata
     3721 cactccgcta gcgctgatgt ccggaatcgt cacacaaaaa accaacacac agatgtaatg
     3781 aaaataaaga tattttattg cggccatcgt gatggctaga gtctaaactc gacctacctc
     3841 cttagcgcgc ggtcgtgtac tgggccggcg gttgccgctg tcggccagca cctcgacgca
     3901 gccgtcggtc catacggctg cgctccggcg agcgatttgt gtgcgaccga cggtgccggc
     3961 cccgctgcgg acgatggcat cgcagcggcc ttgagcccag gcagcatcgt cgaagttgcc
     4021 gtccacgagg ctctggtaga gctgatccag gccgatgcgc agcatgtagg ctcgcagacg
     4081 agggctgccg gccagctcgg gatgccggcg ctcgaagtag cgagtctgct gctccatgca
     4141 agccagccag ggccgccaga agaagatgtt ggccacctcg tactgactgt ccccgaacat
     4201 agcttcggac cagtcgatta cggcggtgat gcggccgttg tcggtcagga cgttgttgct
     4261 gccgaagtcg gcatggacga ggtggcgcac ctcgggacag tcttcggccc acagcatcag
     4321 ttcgtccagg gcttgagcta cgctggcgga cacggtgtcg tccatcacgg tctgccagtg
     4381 gtagacatgg ggatcagcaa tggcgcaaat gaaatcccgc caggttgtgt actggccgat
     4441 gccttggggc ccgaatgggc cgaagccgct ggtttgactg aggtcggcgg cggcaatagc
     4501 atccatagct tcggcgacag gctgtaacac agcaggcagc tctgtttcgg ggaggtcttg
     4561 gagagtaacg ccttgggcgc gtctactgat gcagtatgtc aggctctcgc tgaactcgcc
     4621 gatgtccaac acttcgggga tgggtagtgc agcgctggcg aagtggcggt acacgtagcg
     4681 gtctttgtag aagccatcag cgcagctatt cacccgcagt acatagccgc gtccgccgac
     4741 atcgaagctg aaggctcggc tctcttcgcc ctccgacaac tgcatcaggt cgctcacact
     4801 gtcgaacttc tcgatgagaa atttttcaac gctggtagcg gtgagttcgg gcttcttcat
     4861 ggtggcgcta gtgtcagaag aatcgagctt tttgcaaaag cctaggcctc caaaaaagcc
     4921 tcctcactac ttctggaata gctcagaggc cgaggcggcc tcggcctctg cataaataaa
     4981 aaaaattagt cagccatggg gcggagaatg ggcggaactg ggcggagtta ggggcgggat
     5041 gggcggagtt aggggcggga ctatggttgc tgactaattg agatgcatgc tttgcatact
     5101 tctgcctgct ggggagcctg gggactttcc acacctggtt gctgactaat tgagatgcat
     5161 gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacccta actgacacac
     5221 attccacagc tggttctttc cgcctcagaa ggtacctaac caagttcctc tttcagaggt
     5281 tatttcaggc catggtgctg cgc
//
