LOCUS       Exported                5729 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 5729)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Apr 15, 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..5729
                     /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..2472
                     /label=B4GALT1 (co)
                     /note="B4GALT1 (co)"
     misc_feature    2473..2480
                     /label=PmeI site
                     /note="Used in Flexi Vector Cloning"
     polyA_signal    2609..2830
                     /label=Sv40 late polyadenylation signal
     promoter        2896..2962
                     /label=EM7 bacterial promoter
     CDS             2976..3770
                     /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    3873..4795
                     /label=P15 ori
                     /note="P15 ori"
     misc_feature    4050
                     /label=C2129
                     /note="C2129"
     polyA_signal    complement(4817..4864)
                     /label=Synthetic PolyAdenylation Signal
     misc_feature    complement(4888..5286)
                     /label=BlastR
                     /note="BlastR"
     misc_feature    complement(5311..5729)
                     /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 gagacttcgg gaacccttgc tgtcagggtc agccgctatg
     1321 ccgggagcaa gcttgcagcg cgcttgccgc ctcctggtcg cagtgtgtgc gcttcacctg
     1381 ggagttacac ttgtgtatta tctcgcggga cgcgatctta gcagattgcc tcaactcgta
     1441 ggcgtatcta cccccttgca aggtggtagt aattcagcag ccgctatcgg tcaaagcagt
     1501 ggagagctcc gaacgggggg agctcggcca cctcctccac tcggcgctag ctcccaacct
     1561 cgaccagggg gtgatagcag cccagttgtt gatagtgggc cagggccggc ttcaaacctc
     1621 acctctgtcc ctgttcctca caccaccgcc ctctctcttc cagcctgtcc agaagagagt
     1681 ccattgttgg taggtccaat gcttatagag tttaacatgc ctgtcgacct ggagctcgtg
     1741 gcgaagcaga atcctaacgt aaagatgggg ggccgatacg ctccgcggga ctgtgtcagt
     1801 ccacacaagg tggccataat cattcccttt cgcaacaggc aggaacacct caaatactgg
     1861 ttgtattacc tccatcccgt gcttcagcga cagcagctgg actatgggat atacgtcatt
     1921 aatcaagcgg gtgatacgat tttcaaccgg gccaagctcc tcaacgtagg ttttcaggag
     1981 gcgctcaagg actatgacta cacctgcttc gtcttctctg atgtagacct tattcctatg
     2041 aatgatcata atgcgtacag atgtttttca caacccagac acatatcagt tgctatggat
     2101 aaattcggtt tcagcctgcc atacgttcag tatttcgggg gcgttagcgc actgtcaaag
     2161 cagcaattct tgaccatcaa tggcttccct aataattact gggggtgggg tggggaagat
     2221 gatgatatct ttaatcgctt ggtgtttcgc ggtatgagta tatctcgacc taacgcggtt
     2281 gtgggaaggt gccgcatgat acgccactct agagacaaga aaaatgagcc aaacccgcag
     2341 cgctttgacc ggatcgccca tacaaaggaa actatgttgt ccgatggctt gaatagtctc
     2401 acgtatcaag tgttggatgt ccaaagatac cccctgtaca cacaaattac cgttgacatt
     2461 gggactccta gcgtttaaac gaattcgggc tcggtacccg gggatcctct agagtcgacc
     2521 tgcaggcatg caagctgatc cggctgctaa caaagcccga aaggaagctg agttggctgc
     2581 tgccaccgct gagcaaggcc gcttcgagca gacatgataa gatacattga tgagtttgga
     2641 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
     2701 gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat
     2761 tttatgtttc aggttcaggg ggagatgtgg gaggtttttt taagcaagta aaacctctac
     2821 aaatgtggta aaatcttaat taagaattct aatataactt cgtatagcat acattatacg
     2881 aagttatgga tccactgttg acaattaatc atcggcatag tatatcggca tagtataata
     2941 cgacaaggtg aggaactaaa cccaggaggc agatcatgat tgaacaagat ggattgcacg
     3001 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     3061 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     3121 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt
     3181 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     3241 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     3301 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     3361 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     3421 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     3481 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg
     3541 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     3601 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     3661 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     3721 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga ttcgaaatga
     3781 ccgaccaagc gacgcccaac cggtatcagc tcactcaaag gcggtaatac ggttatccac
     3841 agaatcaggg gataacgcag gaaagaacat gtcaaacatg agaattacaa cttatatcgt
     3901 atggggctga cttcaggtgc tacatttgaa gagataaatt gcactgaaat ctagaaatat
     3961 tttatctgat taataagatg atcttcttga gatcgttttg gtctgcgcgt aatctcttgc
     4021 tctgaaaacg aaaaaaccgc cttgcagggc ggtttttcga aggttctctg agctaccaac
     4081 tctttgaacc gaggtaactg gcttggagga gcgcagtcac caaaacttgt cctttcagtt
     4141 tagccttaac cggcgcatga cttcaagact aactcctcta aatcaattac cagtggctgc
     4201 tgccagtggt gcttttgcat gtctttccgg gttggactca agacgatagt taccggataa
     4261 ggcgcagcgg tcggactgaa cggggggttc gtgcatacag tccagcttgg agcgaactgc
     4321 ctacccggaa ctgagtgtca ggcgtggaat gagacaaacg cggccataac agcggaatga
     4381 caccggtaaa ccgaaaggca ggaacaggag agcgcacgag ggagccgcca gggggaaacg
     4441 cctggtatct ttatagtcct gtcgggtttc gccaccactg atttgagcgt cagatttcgt
     4501 gatgcttgtc aggggggcgg agcctatgga aaaacggctt tgccgcggcc ctctcacttc
     4561 cctgttaagt atcttcctgg catcttccag gaaatctccg ccccgttcgt aagccatttc
     4621 cgctcgccgc agtcgaacga ccgagcgtag cgagtcagtg agcgaggaag cggaatatat
     4681 cctgtatcac atattctgct gacgcaccgg tgcagccttt tttctcctgc cacatgaagc
     4741 acttcactga caccctcatc agtgccaaca tagtaagcca gtatacactc cgctagcgct
     4801 gatgtccgga atcgtcacac aaaaaaccaa cacacagatg taatgaaaat aaagatattt
     4861 tattgcggcc atcgtgatgg ctagagttta gccctcccac acataaccag agggcagcaa
     4921 ttcacgaatc ccaactgccg tcggctgtcc atcactgtcc ttcactatgg ctttgatccc
     4981 aggatgcaga tcgagaagca cctgtcggca ccgtccgcag gggctcaaga tgcccctgtt
     5041 ctcatttccg atcgcgacga tacaagtcag gttgccagct gccgcagcag cagcagtgcc
     5101 cagcaccacg agttctgcac aaggtccccc agtaaaatga tatacattga caccagtgaa
     5161 gatgcggccg tcgctagaga gagctgcgct ggcgacgctg tagtcttcag agatggggat
     5221 gctgttgatt gtagccgttg ctctttcaat gagggtggat tcttcttgag acaaaggctt
     5281 ggccatggtg gcgctagtgt cagaagaatc gagctttttg caaaagccta ggcctccaaa
     5341 aaagcctcct cactacttct ggaatagctc agaggccgag gcggcctcgg cctctgcata
     5401 aataaaaaaa attagtcagc catggggcgg agaatgggcg gaactgggcg gagttagggg
     5461 cgggatgggc ggagttaggg gcgggactat ggttgctgac taattgagat gcatgctttg
     5521 catacttctg cctgctgggg agcctgggga ctttccacac ctggttgctg actaattgag
     5581 atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac accctaactg
     5641 acacacattc cacagctggt tctttccgcc tcagaaggta cctaaccaag ttcctctttc
     5701 agaggttatt tcaggccatg gtgctgcgc
//
