LOCUS       Exported                7799 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 7799)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Friday, May 30, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
COMMENT     pRSG195 designed by Brock F. Binkowski

            

            PCR amplify pF-BiBiTsmN w/ BFBRSG15/BFBRSG14 (PCR #1; 3.9 kbp);

            PCR amplify pF-4B w/ BFBRSG13/BFBRSG16 (PCR #2; 871 bp); 

            2X component Gibson Assembly;

            electroporation of BR610.
FEATURES             Location/Qualifiers
     source          1..7799
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    complement(22..209)
                     /label=SV40\polyA\signal
     misc_feature    complement(213..275)
                     /label=MCS 2
     promoter        276..866
                     /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."
     intron          981..1113
                     /label=chimeric intron
                     /note="Note that this is the actual sequence spliced out. 
                     The 5' donor site includes a 5' GA and the 3' acceptor site
                     adds a 3'G."
     promoter        1157..1176
                     /label=T7 promoter
     CDS             1189..1224
                     /codon_start=1
                     /label=SmBiT
                     /note="SmBiT"
                     /translation="MVTGYRLFEEIL"
     CDS             1225..1269
                     /codon_start=1
                     /label=15GS linker
                     /note="15GS linker"
                     /translation="GSSGGGGSGGGGSSG"
     misc_feature    1270..1277
                     /label=SgfI Site
                     /note="Used in Flexi Vector Cloning"
     misc_feature    1279..4542
                     /label=LATS2
                     /note="LATS2"
     misc_feature    4543..4550
                     /label=PmeI site
                     /note="Used in Flexi Vector Cloning"
     polyA_signal    4679..4900
                     /label=Sv40 late polyadenylation signal
     promoter        4966..5032
                     /label=EM7 bacterial promoter
     CDS             5046..5840
                     /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    5943..6865
                     /label=P15 ori
                     /note="P15 ori"
     misc_feature    6120
                     /label=C2129
                     /note="C2129"
     polyA_signal    complement(6887..6934)
                     /label=Synthetic PolyAdenylation Signal
     misc_feature    complement(6958..7356)
                     /label=BlastR
                     /note="BlastR"
     misc_feature    complement(7381..7799)
                     /label=SV40 promoter
                     /note="SV40 promoter"
ORIGIN
        1 aagacgcgtt aattaagacg tcggcagtga aaaaaatgct ttatttgtga aatttgtgat
       61 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
      121 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac
      181 ctctacaaat gtggtatggc tgattatgat ccgcggccgc aatacgtcga cgttatcagc
      241 tgacttcgta cgagagccta ggattatggc gcgccactag ttattaatag taatcaatta
      301 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      361 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      421 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      481 ctgcccactt ggcagtacat caagtgtatc atatgccaag tccgccccct attgacgtca
      541 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttacgg gactttccta
      601 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      661 acaccaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      721 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaata
      781 accccgcccc gttgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      841 gagctggttt agtgaaccgt cagatcacta gaagctttat tgcggtagtt tatcacagtt
      901 aaattgctaa cgcagtcagt gcttctgaca caacagtctc gaacttaagc tgcagaagtt
      961 ggtcgtgagg cactgggcag gtaagtatca aggttacaag acaggtttaa ggagaccaat
     1021 agaaactggg cttgtcgaga cagagaagac tcttgcgttt ctgataggca cctattggtc
     1081 ttactgacat ccactttgcc tttctctcca caggtgtcca ctcccagttc aattacagct
     1141 cttaaggcta gagtattaat acgactcact atagggctag cgctcaccat ggtgaccggc
     1201 taccggctgt tcgaggagat tctcgggagc tccggtggtg gcgggagcgg aggtggaggc
     1261 tcgagcggtg cgatcgccat gaggccaaag acttttcctg ccacgactta ttctggaaat
     1321 agccggcagc gactgcaaga gattcgtgag gggttaaaac agccatccaa gtcttcggtt
     1381 caggggctac ccgcaggacc aaacagtgac acttccctgg atgccaaagt cctggggagc
     1441 aaagatgcca ccaggcagca gcagcagatg agagccaccc caaagttcgg accttatcag
     1501 aaagccttga gggaaatcag atattccttg ttgccttttg ctaatgaatc gggcacctct
     1561 gcagctgcag aagtgaaccg gcaaatgctg caggaactgg tgaacgcagg atgcgaccag
     1621 gagatggctg gccgagctct caagcagact ggcagcagga gcatcgaggc cgccctggag
     1681 tacatcagca agatgggcta cctggacccg aggaatgagc agattgtgcg ggtcattaag
     1741 cagacctccc caggaaaggg gctcatgcca accccagtga cgcggaggcc cagcttcgaa
     1801 ggaaccggcg attcgtttgc gtcctaccac cagctgagcg gtacccccta cgagggccca
     1861 agcttcggcg ctgacggccc cacggcgctg gaggagatgc cgcggccgta cgtggactac
     1921 cttttccccg gagtcggccc ccacgggccc ggccaccagc accagcaccc acccaagggc
     1981 tacggtgcca gcgtagaggc agcaggggca cacttcccgc tgcagggcgc gcactacggg
     2041 cggccgcacc tgctggtgcc tggggaaccc ctgggctacg gagtgcagcg cagcccctcc
     2101 ttccagagca agacgccgcc ggagaccggg ggttacgcca gcctgcccac gaagggccag
     2161 ggaggaccgc caggcgccgg cctcgctttc ccaccccctg ccgccgggct ctacgtgccg
     2221 cacccacacc acaagcaggc cggtcccgcg gcccaccagc tgcatgtgct gggctcccgc
     2281 agccaggtgt tcgccagcga cagccccccg cagagcctgc tcactccctc gcggaacagc
     2341 ctcaacgtgg acctgtatga attgggcagc acctccgtcc agcagtggcc ggctgccacc
     2401 ctggcccgcc gggactccct gcagaagccg ggcctggagg cgccgccgcg cgcgcacgtg
     2461 gccttccggc ctgactgccc agtgcccagc aggaccaact ccttcaacag ccaccagccg
     2521 cggcccggtc cgcctggcaa ggccgagccc tccctgcccg cccccaacac cgtgacggct
     2581 gtcacggccg cgcacatctt gcacccggtg aagagcgtgc gtgtgctgag gccggagccg
     2641 cagacggctg tggggccctc gcaccccgcc tgggtgcccg cgcctgcccc ggcccccgcc
     2701 cccgcccccg ccccggctgc ggagggcttg gacgccaagg aggagcatgc cctggcgctg
     2761 ggcggcgcag gcgccttccc gctggacgtg gagtacggag gcccagaccg gaggtgcccg
     2821 cctccgccct acccgaagca cctgctgctg cgcagcaagt cggagcagta cgacctggac
     2881 agcctgtgcg caggcatgga gcagagcctc cgtgcgggcc ccaacgagcc cgagggcggc
     2941 gacaagagcc gcaaaagcgc caagggggac aaaggcggaa aggataaaaa gcagattcag
     3001 acctctcccg ttcccgtccg caaaaacagc agagacgaag agaagagaga gtcacgcatc
     3061 aagagctact cgccatacgc ctttaagttc ttcatggagc agcacgtgga gaatgtcatc
     3121 aaaacctacc agcagaaggt taaccggagg ctgcagctgg agcaagaaat ggccaaagct
     3181 ggactctgtg aagctgagca ggagcagatg cggaagatcc tctaccagaa agagtctaat
     3241 tacaacaggt taaagagggc caagatggac aagtctatgt ttgtcaagat caaaaccctg
     3301 gggatcggtg cctttggaga agtgtgcctt gcttgtaagg tggacactca cgccctgtac
     3361 gccatgaaga ccctaaggaa aaaggatgtc ctgaaccgga atcaggtggc ccacgtcaag
     3421 gccgagaggg acatcctggc cgaggcagac aatgagtggg tggtcaaact ctactactcc
     3481 ttccaagaca aagacagcct gtactttgtg atggactaca tccctggtgg ggacatgatg
     3541 agcctgctga tccggatgga ggtcttccct gagcacctgg cccggttcta catcgcagag
     3601 ctgactttgg ccattgagag tgtccacaag atgggcttca tccaccgaga catcaagcct
     3661 gataacattt tgatagatct ggatggtcac attaaactca cagatttcgg cctctgcact
     3721 gggttcaggt ggactcacaa ttccaaatat taccagaaag ggagccatgt cagacaggac
     3781 agcatggagc ccagcgacct ctgggatgat gtgtctaact gtcggtgtgg ggacaggctg
     3841 aagaccctag agcagagggc gcggaagcag caccagaggt gcctggcaca ttcactggtg
     3901 gggactccaa actacatcgc acccgaggtg ctcctccgca aagggtacac tcaactctgt
     3961 gactggtgga gtgttggagt gattctcttc gagatgctgg tggggcagcc gccctttttg
     4021 gcacctactc ccacagaaac ccagctgaag gtgatcaact gggagaacac gctccacatt
     4081 ccagcccagg tgaagctgag ccctgaggcc agggacctca tcaccaagct gtgctgctcc
     4141 gcagaccacc gcctggggcg gaatggggcc gatgacctga aggcccaccc cttcttcagc
     4201 gccattgact tctccagtga catccggaag cagccagccc cctacgttcc caccatcagc
     4261 caccccatgg acacctcgaa tttcgacccc gtagatgaag aaagcccttg gaacgatgcc
     4321 agcgaaggta gcaccaaggc ctgggacaca ctcacctcgc ccaataacaa gcatcctgag
     4381 cacgcatttt acgaattcac cttccgaagg ttctttgatg acaatggcta cccctttcga
     4441 tgcccaaagc cttcaggagc agaagcttca caggctgaga gctcagattt agaaagctct
     4501 gatctggtgg atcagactga aggctgccag cctgtgtacg tggtttaaac gaattcgggc
     4561 tcggtacccg gggatcctct agagtcgacc tgcaggcatg caagctgatc cggctgctaa
     4621 caaagcccga aaggaagctg agttggctgc tgccaccgct gagcaaggcc gcttcgagca
     4681 gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa
     4741 tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat
     4801 aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggagatgtgg
     4861 gaggtttttt taagcaagta aaacctctac aaatgtggta aaatcttaat taagaattct
     4921 aatataactt cgtatagcat acattatacg aagttatgga tccactgttg acaattaatc
     4981 atcggcatag tatatcggca tagtataata cgacaaggtg aggaactaaa cccaggaggc
     5041 agatcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     5101 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     5161 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     5221 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     5281 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     5341 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     5401 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     5461 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     5521 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc
     5581 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     5641 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     5701 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     5761 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     5821 ttcttgacga gttcttctga ttcgaaatga ccgaccaagc gacgcccaac cggtatcagc
     5881 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     5941 gtcaaacatg agaattacaa cttatatcgt atggggctga cttcaggtgc tacatttgaa
     6001 gagataaatt gcactgaaat ctagaaatat tttatctgat taataagatg atcttcttga
     6061 gatcgttttg gtctgcgcgt aatctcttgc tctgaaaacg aaaaaaccgc cttgcagggc
     6121 ggtttttcga aggttctctg agctaccaac tctttgaacc gaggtaactg gcttggagga
     6181 gcgcagtcac caaaacttgt cctttcagtt tagccttaac cggcgcatga cttcaagact
     6241 aactcctcta aatcaattac cagtggctgc tgccagtggt gcttttgcat gtctttccgg
     6301 gttggactca agacgatagt taccggataa ggcgcagcgg tcggactgaa cggggggttc
     6361 gtgcatacag tccagcttgg agcgaactgc ctacccggaa ctgagtgtca ggcgtggaat
     6421 gagacaaacg cggccataac agcggaatga caccggtaaa ccgaaaggca ggaacaggag
     6481 agcgcacgag ggagccgcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     6541 gccaccactg atttgagcgt cagatttcgt gatgcttgtc aggggggcgg agcctatgga
     6601 aaaacggctt tgccgcggcc ctctcacttc cctgttaagt atcttcctgg catcttccag
     6661 gaaatctccg ccccgttcgt aagccatttc cgctcgccgc agtcgaacga ccgagcgtag
     6721 cgagtcagtg agcgaggaag cggaatatat cctgtatcac atattctgct gacgcaccgg
     6781 tgcagccttt tttctcctgc cacatgaagc acttcactga caccctcatc agtgccaaca
     6841 tagtaagcca gtatacactc cgctagcgct gatgtccgga atcgtcacac aaaaaaccaa
     6901 cacacagatg taatgaaaat aaagatattt tattgcggcc atcgtgatgg ctagagttta
     6961 gccctcccac acataaccag agggcagcaa ttcacgaatc ccaactgccg tcggctgtcc
     7021 atcactgtcc ttcactatgg ctttgatccc aggatgcaga tcgagaagca cctgtcggca
     7081 ccgtccgcag gggctcaaga tgcccctgtt ctcatttccg atcgcgacga tacaagtcag
     7141 gttgccagct gccgcagcag cagcagtgcc cagcaccacg agttctgcac aaggtccccc
     7201 agtaaaatga tatacattga caccagtgaa gatgcggccg tcgctagaga gagctgcgct
     7261 ggcgacgctg tagtcttcag agatggggat gctgttgatt gtagccgttg ctctttcaat
     7321 gagggtggat tcttcttgag acaaaggctt ggccatggtg gcgctagtgt cagaagaatc
     7381 gagctttttg caaaagccta ggcctccaaa aaagcctcct cactacttct ggaatagctc
     7441 agaggccgag gcggcctcgg cctctgcata aataaaaaaa attagtcagc catggggcgg
     7501 agaatgggcg gaactgggcg gagttagggg cgggatgggc ggagttaggg gcgggactat
     7561 ggttgctgac taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga
     7621 ctttccacac ctggttgctg actaattgag atgcatgctt tgcatacttc tgcctgctgg
     7681 ggagcctggg gactttccac accctaactg acacacattc cacagctggt tctttccgcc
     7741 tcagaaggta cctaaccaag ttcctctttc agaggttatt tcaggccatg gtgctgcgc
//
