LOCUS       Exported                5273 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 5273)
  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..5273
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    complement(15..202)
                     /label=SV40\polyA\signal
     misc_feature    97..116
                     /label=CP4 TTAACAACAACAATTGCATT
                     /note="CP4 TTAACAACAACAATTGCATT"
     misc_feature    complement(206..255)
                     /label=MCS 2
     gene            complement(278..751)
                     /label=LgBiT
                     /note="LgBiT"
     misc_feature    483..502
                     /label=CP10
                     /note="CP10"
     CDS             complement(752..796)
                     /codon_start=1
                     /label=15GS linker
                     /note="15GS linker"
                     /translation="GSSGGGGSGGGGSSG"
     misc_feature    783..802
                     /label=ECoRC1 site CP11 CCACCTGAACTGCCTTGAGC
                     /note="ECoRC1 site CP11 CCACCTGAACTGCCTTGAGC"
     misc_feature    803..1288
                     /label=MAFG
                     /note="MAFG"
     misc_feature    complement(1290..1297)
                     /label=SgfI Site
                     /note="Used in Flexi Vector Cloning"
     misc_feature    1302..1328
                     /label=B globin leader
                     /note="B globin leader"
     misc_feature    complement(1329..1345)
                     /label=SP6 promoter
                     /note="SP6 promoter"
     misc_feature    complement(1348..1393)
                     /label=Minimal CMV Promoter 2
     misc_feature    1371..1393
                     /label=core
                     /note="core"
     promoter        1403..1957
                     /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."
     misc_feature    complement(1424..1450)
                     /label=CMVR TGGGCTATGAACTAATGACCCCGTAAT
                     /note="CMVR TGGGCTATGAACTAATGACCCCGTAAT"
     promoter        1975..2041
                     /label=EM7 bacterial promoter
     misc_feature    complement(2031..2067)
                     /label=BFBRSG4
                     /note="BFBRSG4"
     misc_feature    2042..2079
                     /label=BFBRSG5
                     /note="BFBRSG5"
     CDS             2055..2849
                     /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    2308..2327
                     /label=PRO CP6
                     /note="PRO CP6"
     misc_feature    2808..2827
                     /label=CP7
                     /note="CP7"
     rep_origin      3004..3040
                     /direction=RIGHT
                     /label=ColE1-derived plasmid replication origin
                     /note="The actual origin is the first base."
     misc_feature    3115..3134
                     /label=CP8
                     /note="CP8"
     misc_feature    3615..3634
                     /label=CP9
                     /note="CP9"
     polyA_signal    complement(3722..3769)
                     /label=Synthetic PolyAdenylation Signal
     CDS             complement(3793..4830)
                     /label=HygroR
                     /note=" Feature found by back translation of amino acid 
                     feature definition."
     misc_feature    complement(4810..4841)
                     /label=BFBRSG9
                     /note="BFBRSG9"
     misc_feature    4820..4854
                     /label=BFBRSG10
                     /note="BFBRSG10"
     misc_feature    complement(4855..5273)
                     /label=SV40 promoter
                     /note="SV40 promoter"
     misc_feature    4981..5000
                     /label=CP 5 TAACTCCGCCCAGTTCCGCC
                     /note="CP 5 TAACTCCGCCCAGTTCCGCC"
     misc_feature    complement(join(5249..5273,1..10))
                     /label=BFBRSG12
                     /note="BFBRSG12"
     misc_feature    join(5264..5273,1..20)
                     /label=BFBRSG11
                     /note="BFBRSG11"
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 gccgatcggg catccgtctt ggactttact attgtgatga
      841 cgctggtggc ggccaccttg cctgggacga ggggccccag gccggcggca agggggcccc
      901 gggctggcgc cacggggctg cgggccaccg tccgggcgaa ggtctgcagc gcctcgtact
      961 tggagcgcag cgcgtcgagc tccagcttca tgctggcgtt ctctgaggcc agcttctcca
     1021 cctcctgctg cagctccgcc ttctgcttct ccagctcctc cttctgcgtc acccgcttca
     1081 cgcggcagct ggcagcgtag ccgcggttct tgagcgtgcg ccggcgctgc ttcagctgga
     1141 cgatctcctc cttggacagg ccccgcaggt gctggttcag ctcccgcacc gacatggtca
     1201 ccagctcctc atccgtcagg ctggtgccat tctcacccgg ctcccgcttc accttcaagg
     1261 ccttgtttcc tttattgggg gtcgtcatgg cgatcgctag ctgcaaaaag aacaagtagc
     1321 ttgtattcta tagtgtcacc taaatgggcg atctgacggt tcactaaacc agctctgctt
     1381 atataggcct ccctggcgcg ccactagtta ttaatagtaa tcaattacgg ggtcattagt
     1441 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
     1501 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
     1561 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
     1621 agtacatcaa gtgtatcata tgccaagtcc gccccctatt gacgtcaatg acggtaaatg
     1681 gcccgcctgg cattatgccc agtacatgac cttacgggac tttcctactt ggcagtacat
     1741 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca ccaatgggcg
     1801 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
     1861 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaataacc ccgccccgtt
     1921 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtcaaa tcgaattcta atactgttga
     1981 caattaatca tcggcatagt atatcggcat agtataatac gacaaggtga ggaactaaac
     2041 ccaggaggca gatcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     2101 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     2161 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     2221 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     2281 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     2341 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     2401 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     2461 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     2521 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     2581 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     2641 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     2701 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     2761 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     2821 tctatcgcct tcttgacgag ttcttctgat tcgaaatgac cgaccaagcg acgcccaacc
     2881 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     2941 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     3001 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     3061 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     3121 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     3181 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     3241 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     3301 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     3361 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     3421 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     3481 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     3541 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ttcaagaaga
     3601 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     3661 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttatagt ccggaatcgt
     3721 cacacaaaaa accaacacac agatgtaatg aaaataaaga tattttattg cggccatcgt
     3781 gatggctaga gtctaaactc gacctacctc cttagcgcgc ggtcgtgtac tgggccggcg
     3841 gttgccgctg tcggccagca cctcgacgca gccgtcggtc catacggctg cgctccggcg
     3901 agcgatttgt gtgcgaccga cggtgccggc cccgctgcgg acgatggcat cgcagcggcc
     3961 ttgagcccag gcagcatcgt cgaagttgcc gtccacgagg ctctggtaga gctgatccag
     4021 gccgatgcgc agcatgtagg ctcgcagacg agggctgccg gccagctcgg gatgccggcg
     4081 ctcgaagtag cgagtctgct gctccatgca agccagccag ggccgccaga agaagatgtt
     4141 ggccacctcg tactgactgt ccccgaacat agcttcggac cagtcgatta cggcggtgat
     4201 gcggccgttg tcggtcagga cgttgttgct gccgaagtcg gcatggacga ggtggcgcac
     4261 ctcgggacag tcttcggccc acagcatcag ttcgtccagg gcttgagcta cgctggcgga
     4321 cacggtgtcg tccatcacgg tctgccagtg gtagacatgg ggatcagcaa tggcgcaaat
     4381 gaaatcccgc caggttgtgt actggccgat gccttggggc ccgaatgggc cgaagccgct
     4441 ggtttgactg aggtcggcgg cggcaatagc atccatagct tcggcgacag gctgtaacac
     4501 agcaggcagc tctgtttcgg ggaggtcttg gagagtaacg ccttgggcgc gtctactgat
     4561 gcagtatgtc aggctctcgc tgaactcgcc gatgtccaac acttcgggga tgggtagtgc
     4621 agcgctggcg aagtggcggt acacgtagcg gtctttgtag aagccatcag cgcagctatt
     4681 cacccgcagt acatagccgc gtccgccgac atcgaagctg aaggctcggc tctcttcgcc
     4741 ctccgacaac tgcatcaggt cgctcacact gtcgaacttc tcgatgagaa atttttcaac
     4801 gctggtagcg gtgagttcgg gcttcttcat ggtggcgcta gtgtcagaag aatcgagctt
     4861 tttgcaaaag cctaggcctc caaaaaagcc tcctcactac ttctggaata gctcagaggc
     4921 cgaggcggcc tcggcctctg cataaataaa aaaaattagt cagccatggg gcggagaatg
     4981 ggcggaactg ggcggagtta ggggcgggat gggcggagtt aggggcggga ctatggttgc
     5041 tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc
     5101 acacctggtt gctgactaat tgagatgcat gctttgcata cttctgcctg ctggggagcc
     5161 tggggacttt ccacacccta actgacacac attccacagc tggttctttc cgcctcagaa
     5221 ggtacctaac caagttcctc tttcagaggt tatttcaggc catggtgctg cgc
//
