LOCUS       sequence-300185-        8379 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses glucagon peptide on mammalian cell surface.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8379)
  AUTHORS   Cho KF, Branon TC, Rajeev S, Svinkina T, Udeshi ND, Thoudam T, Kwak 
            C, Rhee HW, Lee IK, Carr SA, Ting AY
  TITLE     Split-TurboID enables contact-dependent proximity labeling in cells.
  JOURNAL   Proc Natl Acad Sci U S A. 2020 May 18. pii: 1919528117. doi: 
            10.1073/pnas.1919528117.
  PUBMED    32424107
REFERENCE   2  (bases 1 to 8379)
  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: "Proc Natl 
            Acad Sci U S A. 2020 May 18. pii: 1919528117. doi: 
            10.1073/pnas.1919528117."
COMMENT     Sequence Label: pCAG CAG Glucagon-ICAM1-3xHA-pre-mGRASP
FEATURES             Location/Qualifiers
     source          1..8379
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          662..1678
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1602..1624
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1686..1705
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             3317..3406
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     intron          4275..4407
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     4390..4409
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    4416..5004
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4469..4489)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4887..4898)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5031..5048)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5037..5148
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5244..5264)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5244..5262)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5283..5299)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5401..5420)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5466..5521
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5520..5539)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5882..5898)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5906..5922
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5930..5936,5937..5954,5955..5960))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5975..5996
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6055..6250
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6101..6236
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6256..6390
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6293..6312)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6347..6366
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6458..6475)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6629..7217)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6709..6728)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7388..8179,8180..8248))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     8011..8030
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8249..8353)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggsgcgc gccrggsggg gsgggggggg ggggggggsg
      541 gggsggggsg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc
      721 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc
      781 tccgggctgt aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga
      841 aagccttgag gggctccggg agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg
      901 cgtgtgtgtg tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg
      961 ctgcgggcgc ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg
     1021 gggcggtgcc ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg tgcggggtgt
     1081 gtgcgtgggg gggtgagcag ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca
     1141 cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg
     1201 tggcgcgggg ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc
     1261 ggggccgcct cgggccgggg agggctcggg ggaggggcgc ggcggccccc ggagcgccgg
     1321 cggctgtcga ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc
     1381 gcagggactt cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac
     1441 cccctctagc gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga
     1501 gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc
     1561 gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt
     1621 gaccggcggc tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
     1681 cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tcggtaccat
     1741 gagtgcactt ctgatcctag cccttgtggg agctgctgtt gctcattcac agggcacatt
     1801 caccagtgac tacagcaagt atctggactc caagaaggcc caagagtttg tgcagtggtt
     1861 gatgaataca ggtaccggga acggaaatgg caatggtaac gggaatggaa acggcaatgg
     1921 taacgggaat actagtcaga catctgtgtc cccctcaaaa gtcatcctgc cccggggagg
     1981 ctccgtgctg gtgacatgca gcacctcctg tgaccagccc aagttgttgg gcatagagac
     2041 cccgttgcct aaaaaggagt tgctcctgcc tgggaacaac cggaaggtgt atgaactgag
     2101 caatgtgcaa gaagatagcc aaccaatgtg ctattcaaac tgccctgatg ggcagtcaac
     2161 agctaaaacc ttcctcaccg tgtactggac tccagaacgg gtggaactgg cacccctccc
     2221 ctcttggcag ccagtgggca agaaccttac cctacgctgc caggtggagg gtggggcacc
     2281 ccgggccaac ctcaccgtgg tgctgctccg tggggagaag gagctgaaac gggagccagc
     2341 tgtgggggag cccgctgagg tcacgaccac ggtgctggtg aggagagatc accatggagc
     2401 caatttctcg tgccgcactg aactggacct gcggccccaa gggctggagc tgtttgagaa
     2461 cacctcggcc ccctaccagc tccagacctt tgtcctgcca gcgactcccc cacaacttgt
     2521 cagcccccgg gtcctagagg tggacacgca ggggaccgtg gtctgttccc tggacgggct
     2581 gttcccagtc tcggaggccc aggtccacct ggcactgggg gaccagaggt tgaaccccac
     2641 agtcacctat ggcaacgact ccttctcggc caaggcctca gtcagtgtga ccgcagagga
     2701 cgagggcacc cagcggctga cgtgtgcagt aatactgggg aaccagagcc aggagacact
     2761 gcagacagtg accatctaca gctttccggc gcccaacgtg attctgacga agccagaggt
     2821 ctcagaaggg accgaggtga cagtgaagtg tgaggcccac cctagagcca aggtgacgct
     2881 gaatggggtt ccagcccagc cactgggccc gagggcccag ctcctgctga aggccacccc
     2941 agaggacaac gggcgcagct tctcctgctc tgcaaccctg gaggtggccg gccagcttat
     3001 acacaagaac cagacccggg agcttcgtgt cctgtatggc ccccgactgg acgagaggga
     3061 ttgtccggga aactggacgt ggccagaaaa ttcccagcag actccaatgt gccaggcttg
     3121 ggggaaccca ttgcccgagc tcaagtgtct aaaggatggc actttcccac tgcccatcgg
     3181 ggaatcagtg actgtcactc gagatcttga gggcacctac ctctgtcggg ccaggagcac
     3241 tcaaggggag gtcacccgcg aggtgaccgt gaatgtgctc tccccccggt atgaggttaa
     3301 caccggtgcg gccgtttacc catacgatgt tcctgactat gcgggctatc cctatgacgt
     3361 cccggactat gcaggatcct atccatatga cgttccagat tacgctccgg ccgccaccgg
     3421 tcttaaggtc gactttcaga aagctagctc tattgtttat aagaaggagg gggaacaggt
     3481 ggagttcagt ttcccattgg ccttcacagt cgagaagctc acaggatcag gagagttgtg
     3541 gtggcaggcc gaaagagcct cctccagcaa gagttggatc acttttgatc tgaaaaacaa
     3601 agaagtgagc gtgaagagag tcacacagga ccctaagctg cagatgggca agaagttgcc
     3661 tttgcacttg acactccccc aagccctgcc ccagtatgct ggctccggga acctcaccct
     3721 cgccttggag gctaagaccg gaaagttgca tcaggaggtc aacctcgtgg tgatgagggc
     3781 aacacagctg cagaagaacc tgacctgcga agtgtggggt cccacctcac ctaaactgat
     3841 gctgtctctg aagctggaga ataaggaggc taaagtcagt aaaagagaga aggctgtgtg
     3901 ggtccttaac ccggaggcag ggatgtggca gtgtcttctt agcgactctg gacaggtgct
     3961 gctggagagc aacatcaagg tgcttcctac ttggtcaaca ccagtacagc cgatggccct
     4021 gatcgtactc gggggagtcg ccggcctgct tctgtttatc ggcctgggca tcttcttctg
     4081 cgtacgctac aagtatcgca acagggatga gggatcttac cacgttgacg aatctaggaa
     4141 ctatatcagt aacagcgctc agagtaatgg cgccgtggtt aaagagaaac agccctccag
     4201 cgccaaatca gcaaacaaaa acaagaaaaa caaggataaa gagtactacg tacttaagta
     4261 actcgaggcc gcaggtaagt atcaaggtta caagacaggt ttaaggagac caatagaaac
     4321 tgggcttgtc gagacagaga agactcttgc gtttctgata ggcacctatt ggtcttactg
     4381 acatccactt tgcctttctc tccacaggtg tcgacaatca acctctggat tacaaaattt
     4441 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     4501 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     4561 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
     4621 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
     4681 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
     4741 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
     4801 tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg tgttgccacc tggattctgc
     4861 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
     4921 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
     4981 tctccctttg ggccgcctcc ccgcctggaa ttcgagctcg gtacgatcag cctcgactgt
     5041 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     5101 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     5161 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     5221 agacaatagc ccagcttttg ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca
     5281 tggtcatagc tgtttcctgt gtgaaattgt tatccgctaa ttcactcctc aggtgcaggc
     5341 tgcctatcag aaggtggtgg ctggtgtggc caatgccctg gctcacaaat accactgaga
     5401 tctttttccc tctgccaaaa attatgggga catcatgaag ccccttgagc atctgacttc
     5461 tggctaataa aggaaattta ttttcattgc aatagtgtgt tggaattttt tgtgtctctc
     5521 actcggaagg acatatggga gggcaaatca tttaaaacat cagaatgagt atttggttta
     5581 gagtttggca acatatgccc atatgctggc tgccatgaac aaaggttggc tataaagagg
     5641 tcatcagtat atgaaacagc cccctgctgt ccattcctta ttccatagaa aagccttgac
     5701 ttgaggttag atttttttta tattttgttt tgtgttattt ttttctttaa catccctaaa
     5761 attttcctta catgttttac tagccagatt tttcctcctc tcctgactac tcccagtcat
     5821 agctgtccct cttctcttat ggagatccct cgacctgcag cccaagcttg gcgtaatcat
     5881 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     5941 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     6001 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagcgg atccgcatct
     6061 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     6121 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     6181 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     6241 cttttgcaaa aagctaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     6301 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     6361 ctcatcaatg tatcttatca tgtctggatc cgctgcatta atgaatcggc caacgcgcgg
     6421 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     6481 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     6541 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     6601 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     6661 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     6721 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     6781 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     6841 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     6901 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     6961 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     7021 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     7081 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     7141 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     7201 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     7261 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     7321 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     7381 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     7441 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     7501 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     7561 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     7621 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     7681 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     7741 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     7801 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     7861 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     7921 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     7981 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     8041 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     8101 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     8161 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     8221 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     8281 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     8341 taggggttcc gcgcacattt ccccgaaaag tgccacctg
//
