LOCUS       Exported                5691 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 5691)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Apr 15, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
COMMENT     pRSG194 designed by Brock F. Binkowski

            

            PCR amplify pF-BiBiTsmC 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..5691
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          1185..2622
                     /label=GLP1R-SmBiT (pFC220K)
     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
     misc_feature    1180..1187
                     /label=SgfI Site
                     /note="Used in Flexi Vector Cloning"
     CDS             1189..2577
                     /codon_start=1
                     /label=GLP1R
                     /translation="MAGAPGPLRLALLLLGMVGRAGPRPQGATVSLWETVQKWREYRRQ
                     CQRSLTEDPPPATDLFCNRTFDEYACWPDGEPGSFVNVSCPWYLPWASSVPQGHVYRFC
                     TAEGLWLQKDNSSLPWRDLSECEESKRGERSSPEEQLLFLYIIYTVGYALSFSALVIAS
                     AILLGFRHLHCTRNYIHLNLFASFILRALSVFIKDAALKWMYSTAAQQHQWDGLLSYQD
                     SLSCRLVFLLMQYCVAANYYWLLVEGVYLYTLLAFSVLSEQWIFRLYVSIGWGVPLLFV
                     VPWGIVKYLYEDEGCWTRNSNMNYWLIIRLPILFAIGVNFLIFVRVICIVVSKLKANLM
                     CKTDIKCRLAKSTLTLIPLLGTHEVIFAFVMDEHARGTLRFIKLFTELSFTSFQGLMVA
                     ILYCFVNNEVQLEFRKSWERWRLEHLHIQRDSSMKPLKCPTSSLSSGATAGSSMYTATC
                     QASCS"
     CDS             2587..2631
                     /codon_start=1
                     /label=15GS linker
                     /translation="GSSGGGGSGGGGSSG"
     CDS             2632..2664
                     /codon_start=1
                     /label=SmBiT
                     /note="SmBiT"
                     /translation="VTGYRLFEEIL"
     polyA_signal    2745..2966
                     /label=Sv40 late polyadenylation signal
     misc_feature    complement(2990..3023)
                     /label=Cre site
                     /note="Cre site"
     promoter        3032..3098
                     /label=EM7 bacterial promoter
     CDS             3112..3906
                     /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      4061..4097
                     /direction=RIGHT
                     /label=ColE1-derived plasmid replication origin
                     /note="The actual origin is the first base."
     polyA_signal    complement(4779..4826)
                     /label=Synthetic PolyAdenylation Signal
     misc_feature    complement(4850..5248)
                     /label=BlastR
                     /note="BlastR"
     misc_feature    complement(5273..5691)
                     /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 cgatcgccat ggccggcgcc
     1201 cccggcccgc tgcgccttgc gctgctgctg ctcgggatgg tgggcagggc cggcccccgc
     1261 ccccagggtg ccactgtgtc cctctgggag acggtgcaga aatggcgaga ataccgacgc
     1321 cagtgccagc gctccctgac tgaggatcca cctcctgcca cagacttgtt ctgcaaccgg
     1381 accttcgatg aatacgcctg ctggccagat ggggagccag gctcgttcgt gaatgtcagc
     1441 tgcccctggt acctgccctg ggccagcagt gtgccgcagg gccacgtgta ccggttctgc
     1501 acagctgaag gcctctggct gcagaaggac aactccagcc tgccctggag ggacttgtcg
     1561 gagtgcgagg agtccaagcg aggggaaaga agctccccgg aggagcagct cctgttcctc
     1621 tacatcatct acacggtggg ctacgcactc tccttctctg ctctggttat cgcctctgcg
     1681 atcctcctcg gcttcagaca cctgcactgc accaggaact acatccacct gaacctgttt
     1741 gcatccttca tcctgcgagc attgtccgtc ttcatcaagg acgcagccct gaagtggatg
     1801 tatagcacag ccgcccagca gcaccagtgg gatgggctcc tctcctacca ggactctctg
     1861 agctgccgcc tggtgtttct gctcatgcag tactgtgtgg cggccaatta ctactggctc
     1921 ttggtggagg gcgtgtacct gtacacactg ctggccttct cggtcttatc tgagcaatgg
     1981 atcttcaggc tctacgtgag cataggctgg ggtgttcccc tgctgtttgt tgtcccctgg
     2041 ggcattgtca agtacctcta tgaggacgag ggctgctgga ccaggaactc caacatgaac
     2101 tactggctca ttatccggct gcccattctc tttgccattg gggtgaactt cctcatcttt
     2161 gttcgggtca tctgcatcgt ggtatccaaa ctgaaggcca atctcatgtg caagacagac
     2221 atcaaatgca gacttgccaa gtccacgctg acactcatcc ccctgctggg gactcatgag
     2281 gtcatctttg cctttgtgat ggacgagcac gcccggggga ccctgcgctt catcaagctg
     2341 tttacagagc tctccttcac ctccttccag gggctgatgg tggccatatt atactgcttt
     2401 gtcaacaatg aggtccagct ggaatttcgg aagagctggg agcgctggcg gcttgagcac
     2461 ttgcacatcc agagggacag cagcatgaag cccctcaagt gtcccaccag cagcctgagc
     2521 agtggagcca cggcgggcag cagcatgtac acagccactt gccaggcctc ctgcagcgtt
     2581 tctcaaggca gttcaggtgg tggcgggagc ggaggtggag gctcgagcgg tgtgaccggc
     2641 taccggctgt tcgaggagat tctgtaaggc cgcgactcta gagtcgacct gcaggcatgc
     2701 aagctgattt ggctgctgcc accgctgagc aaggccgctt cgagcagaca tgataagata
     2761 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
     2821 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa
     2881 caacaattgc attcatttta tgtttcaggt tcagggggag atgtgggagg tttttttaag
     2941 caagtaaaac ctctacaaat gtggtaaaat cttaattaag aattctaata taacttcgta
     3001 tagcatacat tatacgaagt tatggatcca ctgttgacaa ttaatcatcg gcatagtata
     3061 tcggcatagt ataatacgac aaggtgagga actaaaccca ggaggcagat catgattgaa
     3121 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     3181 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     3241 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     3301 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     3361 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     3421 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     3481 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     3541 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     3601 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     3661 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     3721 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     3781 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     3841 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     3901 ttctgattcg aaatgaccga ccaagcgacg cccaaccggt atcagctcac tcaaaggcgg
     3961 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     4021 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     4081 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     4141 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     4201 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     4261 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     4321 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     4381 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     4441 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     4501 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     4561 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     4621 agcagattac gcgcagaaaa aaaggatttc aagaagatcc tttgatcttt tctacggggt
     4681 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     4741 ggatcttcac ctagatcctt ttatagtccg gaatcgtcac acaaaaaacc aacacacaga
     4801 tgtaatgaaa ataaagatat tttattgcgg ccatcgtgat ggctagagtt tagccctccc
     4861 acacataacc agagggcagc aattcacgaa tcccaactgc cgtcggctgt ccatcactgt
     4921 ccttcactat ggctttgatc ccaggatgca gatcgagaag cacctgtcgg caccgtccgc
     4981 aggggctcaa gatgcccctg ttctcatttc cgatcgcgac gatacaagtc aggttgccag
     5041 ctgccgcagc agcagcagtg cccagcacca cgagttctgc acaaggtccc ccagtaaaat
     5101 gatatacatt gacaccagtg aagatgcggc cgtcgctaga gagagctgcg ctggcgacgc
     5161 tgtagtcttc agagatgggg atgctgttga ttgtagccgt tgctctttca atgagggtgg
     5221 attcttcttg agacaaaggc ttggccatgg tggcgctagt gtcagaagaa tcgagctttt
     5281 tgcaaaagcc taggcctcca aaaaagcctc ctcactactt ctggaatagc tcagaggccg
     5341 aggcggcctc ggcctctgca taaataaaaa aaattagtca gccatggggc ggagaatggg
     5401 cggaactggg cggagttagg ggcgggatgg gcggagttag gggcgggact atggttgctg
     5461 actaattgag atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac
     5521 acctggttgc tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg
     5581 gggactttcc acaccctaac tgacacacat tccacagctg gttctttccg cctcagaagg
     5641 tacctaacca agttcctctt tcagaggtta tttcaggcca tggtgctgcg c
//
