LOCUS       Exported                5345 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 5345)
  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..5345
                     /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..754)
                     /label=LgBiT
                     /note="LgBiT"
     misc_feature    complement(755..802)
                     /label=Geneious type: polylinker
                     /note="Geneious type: polylinker"
     gene            complement(803..925)
                     /label=HHR23A UBA1
     misc_feature    complement(926..946)
                     /label=Geneious type: polylinker
                     /note="Geneious type: polylinker"
     gene            complement(947..1069)
                     /label=HHR23A UBA1
     misc_feature    complement(1070..1090)
                     /label=Geneious type: polylinker
                     /note="Geneious type: polylinker"
     gene            complement(1091..1213)
                     /label=HHR23A UBA1
     misc_feature    complement(1214..1234)
                     /label=Geneious type: polylinker
                     /note="Geneious type: polylinker"
     gene            complement(1235..1357)
                     /label=HHR23A UBA1
     promoter        complement(1358..1360)
                     /label=START
     misc_feature    complement(1362..1369)
                     /label=SgfI Site
                     /note="Used in Flexi Vector Cloning"
     misc_feature    1374..1400
                     /label=B globin leader
                     /note="B globin leader"
     misc_feature    complement(1401..1417)
                     /label=SP6 promoter
                     /note="SP6 promoter"
     misc_feature    complement(1420..1465)
                     /label=Minimal CMV Promoter
     misc_feature    1443..1465
                     /label=core
                     /note="core"
     promoter        1475..2029
                     /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        2047..2113
                     /label=EM7 bacterial promoter
     CDS             2127..2921
                     /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      3076..3112
                     /direction=RIGHT
                     /label=ColE1-derived plasmid replication origin
                     /note="The actual origin is the first base."
     polyA_signal    complement(3794..3841)
                     /label=Synthetic PolyAdenylation Signal
     CDS             complement(3865..4902)
                     /label=HygroR
                     /note=" Feature found by back translation of amino acid 
                     feature definition."
     misc_feature    complement(4927..5345)
                     /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 ccatgccaga acctcctccc ccggatcctc
      781 ctccgccgga ccctccacca ccgcctgtca gcaaatactc gacagctcga tgaggattgt
      841 tataggaggc ccgcagggca gctacaacgc gctcgcgttc gtaacccatt gacattatct
      901 cagtaagcat agtctcgtat tcggatccgc tgccacttcc gctcccaccg gtcagcaggt
      961 attcaaccgc cctatgcgga ttattatatg aggctcgcag cgcagcgacc actctctccc
     1021 tctcgtatcc catactcata atctcggtga gcatcgtctc atattcactg ccgctacctg
     1081 atccactacc acccgtcagc agatactcaa cagctctatg ggggttatta taggaagctc
     1141 ggagcgctgc gacaactcgt tcccgttcat agcccatgga cataatctct gtcagcatgg
     1201 tctcatactc tgaacctgac ccgcttccac tccctcctgt cagaagatat tccaccgctc
     1261 ggtgaggatt gttgtagctc gcgcgcagag ccgccacaac gcgctcccgt tcatatccca
     1321 ttgacattat ttccgtgagc atggtttcat attcggacat ggcgatcgct agctgcaaaa
     1381 agaacaagta gcttgtattc tatagtgtca cctaaatggg cgatctgacg gttcactaaa
     1441 ccagctctgc ttatataggc ctccctggcg cgccactagt tattaatagt aatcaattac
     1501 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
     1561 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
     1621 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
     1681 tgcccacttg gcagtacatc aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa
     1741 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttacggg actttcctac
     1801 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta
     1861 caccaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
     1921 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa
     1981 ccccgccccg ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtca aatcgaattc
     2041 taatactgtt gacaattaat catcggcata gtatatcggc atagtataat acgacaaggt
     2101 gaggaactaa acccaggagg cagatcatga ttgaacaaga tggattgcac gcaggttctc
     2161 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     2221 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     2281 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca
     2341 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     2401 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     2461 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     2521 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     2581 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     2641 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     2701 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     2761 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     2821 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     2881 agcgcatcgc cttctatcgc cttcttgacg agttcttctg attcgaaatg accgaccaag
     2941 cgacgcccaa ccggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     3001 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     3061 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     3121 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     3181 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     3241 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     3301 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     3361 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     3421 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     3481 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     3541 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     3601 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     3661 atttcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     3721 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttata
     3781 gtccggaatc gtcacacaaa aaaccaacac acagatgtaa tgaaaataaa gatattttat
     3841 tgcggccatc gtgatggcta gagtctaaac tcgacctacc tccttagcgc gcggtcgtgt
     3901 actgggccgg cggttgccgc tgtcggccag cacctcgacg cagccgtcgg tccatacggc
     3961 tgcgctccgg cgagcgattt gtgtgcgacc gacggtgccg gccccgctgc ggacgatggc
     4021 atcgcagcgg ccttgagccc aggcagcatc gtcgaagttg ccgtccacga ggctctggta
     4081 gagctgatcc aggccgatgc gcagcatgta ggctcgcaga cgagggctgc cggccagctc
     4141 gggatgccgg cgctcgaagt agcgagtctg ctgctccatg caagccagcc agggccgcca
     4201 gaagaagatg ttggccacct cgtactgact gtccccgaac atagcttcgg accagtcgat
     4261 tacggcggtg atgcggccgt tgtcggtcag gacgttgttg ctgccgaagt cggcatggac
     4321 gaggtggcgc acctcgggac agtcttcggc ccacagcatc agttcgtcca gggcttgagc
     4381 tacgctggcg gacacggtgt cgtccatcac ggtctgccag tggtagacat ggggatcagc
     4441 aatggcgcaa atgaaatccc gccaggttgt gtactggccg atgccttggg gcccgaatgg
     4501 gccgaagccg ctggtttgac tgaggtcggc ggcggcaata gcatccatag cttcggcgac
     4561 aggctgtaac acagcaggca gctctgtttc ggggaggtct tggagagtaa cgccttgggc
     4621 gcgtctactg atgcagtatg tcaggctctc gctgaactcg ccgatgtcca acacttcggg
     4681 gatgggtagt gcagcgctgg cgaagtggcg gtacacgtag cggtctttgt agaagccatc
     4741 agcgcagcta ttcacccgca gtacatagcc gcgtccgccg acatcgaagc tgaaggctcg
     4801 gctctcttcg ccctccgaca actgcatcag gtcgctcaca ctgtcgaact tctcgatgag
     4861 aaatttttca acgctggtag cggtgagttc gggcttcttc atggtggcgc tagtgtcaga
     4921 agaatcgagc tttttgcaaa agcctaggcc tccaaaaaag cctcctcact acttctggaa
     4981 tagctcagag gccgaggcgg cctcggcctc tgcataaata aaaaaaatta gtcagccatg
     5041 gggcggagaa tgggcggaac tgggcggagt taggggcggg atgggcggag ttaggggcgg
     5101 gactatggtt gctgactaat tgagatgcat gctttgcata cttctgcctg ctggggagcc
     5161 tggggacttt ccacacctgg ttgctgacta attgagatgc atgctttgca tacttctgcc
     5221 tgctggggag cctggggact ttccacaccc taactgacac acattccaca gctggttctt
     5281 tccgcctcag aaggtaccta accaagttcc tctttcagag gttatttcag gccatggtgc
     5341 tgcgc
//
