LOCUS       sequence-471845-        5691 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express GLP-1R-SmBiT in Mammalian Cells under a CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5691)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5691)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: GLP-1R-SmBiT Fusion Vector
FEATURES             Location/Qualifiers
     source          1..5691
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    24..105
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     enhancer        338..641
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        642..845
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     796..816
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          981..1113
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1096..1115
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1157..1176
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1157..1175
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    2754..2875
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2765..2784)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(2990..3023)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3033..3080
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3112..3906
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3166..3185)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3776..3795
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4064..4652)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4144..4163)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4778..4826
                     /note="synthetic polyadenylation signal"
     CDS             complement(4850..5248)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5245..5254
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5279..5636)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5293..5428
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5445..5636
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5593..5613
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
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
//
