LOCUS       sequence-430133-        7386 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Adhesion G Protein-Coupled Receptor 
            transmembrane domains.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7386)
  AUTHORS   Dates AN, Jones DTD, Smith JS, Skiba MA, Rich MF, Burruss MM, Kruse 
            AC, Blacklow SC
  TITLE     Heterogeneity of Tethered Agonist Signaling in Adhesion G 
            Protein-Coupled Receptors
  JOURNAL   bioRxiv 2022.07.14.500097
REFERENCE   2  (bases 1 to 7386)
  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: "bioRxiv 
            2022.07.14.500097"
COMMENT     Sequence Label: HA-MBP-TEVpCS-ADGRB2 CTF-FLAG
FEATURES             Location/Qualifiers
     source          1..7386
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        49..260
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             273..541
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     sig_peptide     591..650
                     /codon_start=1
                     /product="human interleukin-2 signal sequence"
                     /label=IL-2 signal sequence
                     /translation="MYRMQLLSCIALSLALVTNS"
     CDS             651..677
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1785..1805
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             3831..3854
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    4569..4690
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4580..4599)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4634..4653
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     polyA_signal    4794..5188
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     CDS             complement(5202..5576)
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     enhancer        6169..6472
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(6552..7140)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6632..6651)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    7248..7296
                     /note="synthetic polyadenylation signal"
     misc_feature    join(7307..7386,1..12)
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
ORIGIN
        1 gaacatttct ctatcgaagg atctgcgatc gctccggtgc ccgtcagtgg gcagagcgca
       61 catcgcccac agtccccgag aagttggggg gaggggtcgg caattgaacg ggtgcctaga
      121 gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg
      181 agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg
      241 ggtttgccgc cagaacacag ctgaagcttc gaggggctcg catctctcct tcacgcgccc
      301 gccgccctac ctgaggccgc catccacgcc ggttgagtcg cgttctgccg cctcccgcct
      361 gtggtgcctc ctgaactgcg tccgccgtct aggtaagttt aaagctcagg tcgagaccgg
      421 gcctttgtcc ggcgctccct tggagcctac ctagactcag ccggctctcc acgctttgcc
      481 tgaccctgct tgctcaactc tacgtctttg tttcgttttc tgttctgcgc cgttacagat
      541 ccaagctgtg accggcgcct acctgagatc accggcgaag gagggccacc atgtacagga
      601 tgcaactcct gtcttgcatt gcactaagtc ttgcacttgt cacgaattcg tacccatacg
      661 atgttccaga ttacgctgga tccaaaattg aagaaggcaa actggtgatt tggattaacg
      721 gcgataaagg ctataacggc ctggcggaag tgggcaaaaa atttgaaaaa gataccggca
      781 ttaaagtgac cgtggaacat ccggataaac tggaagaaaa atttccgcag gtggcggcga
      841 ccggcgatgg cccggatatt attttttggg cgcatgatcg ctttggcggc tatgcgcaga
      901 gcggcctgct ggcggaaatt accccggata aagcgtttca ggataaactg tatccgttta
      961 cctgggatgc ggtgcgctat aacggcaaac tgattgcgta tccgattgcg gtggaagcgc
     1021 tgagcctgat ttataacaaa gatctgctgc cgaacccgcc gaaaacctgg gaagaaattc
     1081 cggcgctgga taaagaactg aaagcgaaag gcaaaagcgc gctgatgttt aacctgcagg
     1141 aaccgtattt tacctggccg ctgattgcgg cggatggcgg ctatgcgttt aaatatgaaa
     1201 acggcaaata tgatattaaa gatgtgggcg tggataacgc gggcgcgaaa gcgggcctga
     1261 cctttctggt ggatctgatt aaaaacaaac atatgaacgc ggataccgat tatagcattg
     1321 cggaagcggc gtttaacaaa ggcgaaaccg cgatgaccat taacggcccg tgggcgtgga
     1381 gcaacattga taccagcaaa gtgaactatg gcgtgaccgt gctgccgacc tttaaaggcc
     1441 agccgagcaa accgtttgtg ggcgtgctga gcgcgggcat taacgcggcg agcccgaaca
     1501 aagaactggc gaaagaattt ctggaaaact atctgctgac cgatgaaggc ctggaagcgg
     1561 tgaacaaaga taaaccgctg ggcgcggtgg cgctgaaaag ctatgaagaa gaactggcga
     1621 aagatccgcg cattgcggcg accatggaaa acgcgcagaa aggcgaaatt atgccgaaca
     1681 ttccgcagat gagcgcgttt tggtatgcgg tgcgcaccgc ggtgattaac gcggcgagcg
     1741 gccgccagac cgtggatgaa gcgctgaaag atgcgcagac caacgagaac ctttacttcc
     1801 aatctacatt cgccgtactc gctcagccgc ccaaggacct gacgctggag ctggccggca
     1861 gccccagcgt cccgctcgtc atcggctgtg cagtgtcttg tatggcgttg ctcaccctcc
     1921 tggccattta tgcggctttc tggcgcttta tcaagtccga gcgttctatc attctcctga
     1981 acttctgtct tagtatcctg gcctccaaca tcctcattct cgtgggccag agtcgcgtcc
     2041 tgagtaaagg cgtatgtact atgactgctg cgttcctcca cttctttttc ctcagcagtt
     2101 tctgctgggt gctcaccgaa gcatggcagt cttacctggc cgtgatcggc cgtatgagaa
     2161 cacgcctggt gcgcaagcgc tttctgtgtc tgggctgggg cctgccagct ctggtggtag
     2221 ctgtgagcgt ggggtttaca cgcacaaagg gctacggcac ctccagctac tgttggctga
     2281 gcctggaggg agggctgctc tatgcattcg taggcccggc cgctgtgatc gtcctggtca
     2341 acatgctcat cggtattatc gtcttcaata aactgatggc ccgggatggg atctctgaca
     2401 agtccaagaa acagagggca gggagtgaga ggtgcccctg ggcgtctctg cttctgcctt
     2461 gtagtgcctg cggggctgtt ccaagccccc tgcttagctc tgcttctgct agaaacgcga
     2521 tggccagcct gtggagctct tgcgtggtcc tgccactcct ggctctgact tggatgagtg
     2581 ccgttcttgc gatgactgac aggcgctcag tgctgttcca ggctctgttc gcagtattca
     2641 attccgccca gggctttgtg atcaccgcag ttcactgctt cctccgccgt gaggtgcagg
     2701 acgtcgtgaa atgtcagatg ggcgtgtgca gagccgatga atccgaggat tcccccgaca
     2761 gctgcaagaa cggacagctg cagatcctgt ccgacttcga gaaagatgtt gaccttgcct
     2821 gccagacagt tctgttcaag gaggtgaata cttgtaaccc gagtaccatc actgggactc
     2881 tgtccaggct gtccctggat gaagacgaag agccaaagtc ttgtcttgtg ggccccgagg
     2941 gaagcctgtc cttctcaccc ttgcccggca acatccttgt acccatggcc gctagccctg
     3001 gccttggtga gccccctccc cctcaggaag ccaacccggt gtatatgtgc ggcgagggtg
     3061 gcctccgcca attggacctg acctggctgc gtcctacgga accaggcagc gagggcgatt
     3121 acatggttct gcctagacgg actctgagcc tgcagcccgg aggcggtgga ggcggtggag
     3181 aggacgcacc acgggctagg ccggaaggca caccgagacg cgcggccaag actgtcgctc
     3241 atacagaggg gtatccatcc tttctttccg tggaccattc cgggctgggc ttgggaccgg
     3301 cttacggttc cttgcagaac ccttacggca tgacgtttca gccacctccc cctaccccat
     3361 ccgccagaca ggttcctgag cctggcgaga ggtccagaac aatgccacgg accgtacccg
     3421 ggtccacaat gaagatggga agtcttgaaa gaaagaaact gcgctacagt gacctggatt
     3481 tcgagaaggt gatgcatacc cgcaaaagac actccgagct gtatcatgag ctgaaccaga
     3541 aatttcacac attcgaccgg taccggtccc agtccaccgc caagcgggag aagcgctggt
     3601 ctgtcagctc tggaggtgct gccgagcgta gcgtatgtac cgataaacca agccctggag
     3661 agcgcccctc cctgtcccag cacaggcgcc atcagagctg gagcacattt aaatccatga
     3721 ctttgggtag cttgccacct aagccacgcg aacgtctgac gctgcaccgt gctgccgcgt
     3781 gggagcctac cgaaccgcca gacggtgatt ttcaaaccga ggttggttct gactacaaag
     3841 acgatgacga caagtgaaga tctgacaaaa ctcacacatg cccaccgtgc ccagcacctg
     3901 aactcctggg gggaccgtca gtcttcctct tccccccaaa acccaaggac accctcatga
     3961 tctcccggac ccctgaggtc acatgcgtgg tggtggacgt gagccacgaa gaccctgagg
     4021 tcaagttcaa ctggtacgtg gacggcgtgg aggtgcataa tgccaagaca aagccgcggg
     4081 aggagcagta caacagcacg taccgtgtgg tcagcgtcct caccgtcctg caccaggact
     4141 ggctgaatgg caaggagtac aagtgcaagg tctccaacaa agccctccca gcccccatcg
     4201 agaaaaccat ctccaaagcc aaagggcagc cccgagaacc acaggtgtac accctgcccc
     4261 catcccggga ggagatgacc aagaaccagg tcagcctgac ctgcctggtc aaaggcttct
     4321 atcccagcga catcgccgtg gagtgggaga gcaatgggca gccggagaac aactacaaga
     4381 ccacgcctcc cgtgctggac tccgacggct ccttcttcct ctacagcaag ctcaccgtgg
     4441 acaagagcag gtggcagcag gggaacgtct tctcatgctc cgtgatgcac gaggctctgc
     4501 acaaccacta cacgcagaag agcctctccc tgtctccggg taaatgagtg ctagctggcc
     4561 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     4621 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     4681 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg
     4741 ggaggttttt taaagcaagt aaaacctcta caaatgtggt atggaattaa ttctaaaata
     4801 cagcatagca aaactttaac ctccaaatca agcctctact tgaatccttt tctgagggat
     4861 gaataaggca taggcatcag gggctgttgc caatgtgcat tagctgtttg cagcctcacc
     4921 ttctttcatg gagtttaaga tatagtgtat tttcccaagg tttgaactag ctcttcattt
     4981 ctttatgttt taaatgcact gacctcccac attccctttt tagtaaaata ttcagaaata
     5041 atttaaatac atcattgcaa tgaaaataaa tgttttttat taggcagaat ccagatgctc
     5101 aaggcccttc ataatatccc ccagtttagt agttggactt agggaacaaa ggaaccttta
     5161 atagaaattg gacagcaaga aagcgagctt ctagcttatc ctcagtcctg ctcctctgcc
     5221 acaaagtgca cgcagttgcc ggccgggtcg cgcagggcga actcccgccc ccacggctgc
     5281 tcgccgatct cggtcatggc cggcccggag gcgtcccgga agttcgtgga cacgacctcc
     5341 gaccactcgg cgtacagctc gtccaggccg cgcacccaca cccaggccag ggtgttgtcc
     5401 ggcaccacct ggtcctggac cgcgctgatg aacagggtca cgtcgtcccg gaccacaccg
     5461 gcgaagtcgt cctccacgaa gtcccgggag aacccgagcc ggtcggtcca gaactcgacc
     5521 gctccggcga cgtcgcgcgc ggtgagcacc ggaacggcac tggtcaactt ggccatgatg
     5581 gctcctcctg tcaggagagg aaagagaaga aggttagtac aattgctata gtgagttgta
     5641 ttatactatg cagatatact atgccaatga ttaattgtca aactagggct gcagggttca
     5701 tagtgccact tttcctgcac tgccccatct cctgcccacc ctttcccagg catagacagt
     5761 cagtgactta ccaaactcac aggagggaga aggcagaagc ttgagacaga cccgcgggac
     5821 cgccgaactg cgaggggacg tggctagggc ggcttctttt atggtgcgcc ggccctcgga
     5881 ggcagggcgc tcggggaggc ctagcggcca atctgcggtg gcaggaggcg gggccgaagg
     5941 ccgtgcctga ccaatccgga gcacatagga gtctcagccc cccgccccaa agcaagggga
     6001 agtcacgcgc ctgtagcgcc agcgtgttgt gaaatggggg cttggggggg ttggggccct
     6061 gactagtcaa aacaaactcc cattgacgtc aatggggtgg agacttggaa atccccgtga
     6121 gtcaaaccgc tatccacgcc cattgatgta ctgccaaaac cgcatcatca tggtaatagc
     6181 gatgactaat acgtagatgt actgccaagt aggaaagtcc cataaggtca tgtactgggc
     6241 ataatgccag gcgggccatt taccgtcatt gacgtcaata gggggcgtac ttggcatatg
     6301 atacacttga tgtactgcca agtgggcagt ttaccgtaaa tactccaccc attgacgtca
     6361 atggaaagtc cctattggcg ttactatggg aacatacgtc attattgacg tcaatgggcg
     6421 ggggtcgttg ggcggtcagc caggcgggcc atttaccgta agttatgtaa cgcctgcagg
     6481 ttaattaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     6541 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     6601 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     6661 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     6721 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     6781 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     6841 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     6901 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     6961 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     7021 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     7081 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     7141 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     7201 gggattttgg tcatggctag ttaattaaca tttaaatcag cggccgcaat aaaatatctt
     7261 tattttcatt acatctgtgt gttggttttt tgtgtgaatc gtaactaaca tacgctctcc
     7321 atcaaaacaa aacgaaacaa aacaaactag caaaataggc tgtccccagt gcaagtgcag
     7381 gtgcca
//
