LOCUS       sequence-430004-        6195 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 6195)
  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 6195)
  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-ADGRE4 CTFdeltaTA-FLAG
FEATURES             Location/Qualifiers
     source          1..6195
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        31..242
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             255..523
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     sig_peptide     573..632
                     /codon_start=1
                     /product="human interleukin-2 signal sequence"
                     /label=IL-2 signal sequence
                     /translation="MYRMQLLSCIALSLALVTNS"
     CDS             633..659
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1767..1787
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQA"
     CDS             2622..2645
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    3360..3481
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3371..3390)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3425..3444
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     polyA_signal    3585..3979
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     CDS             complement(3993..4367)
                     /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        4960..5263
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(5343..5931)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5423..5442)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    6039..6087
                     /note="synthetic polyadenylation signal"
     misc_feature    6098..6189
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     6138..6157
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggatctgcga tcgctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg
       61 agaagttggg gggaggggtc ggcaattgaa cgggtgccta gagaaggtgg cgcggggtaa
      121 actgggaaag tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt
      181 atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac
      241 agctgaagct tcgaggggct cgcatctctc cttcacgcgc ccgccgccct acctgaggcc
      301 gccatccacg ccggttgagt cgcgttctgc cgcctcccgc ctgtggtgcc tcctgaactg
      361 cgtccgccgt ctaggtaagt ttaaagctca ggtcgagacc gggcctttgt ccggcgctcc
      421 cttggagcct acctagactc agccggctct ccacgctttg cctgaccctg cttgctcaac
      481 tctacgtctt tgtttcgttt tctgttctgc gccgttacag atccaagctg tgaccggcgc
      541 ctacctgaga tcaccggcga aggagggcca ccatgtacag gatgcaactc ctgtcttgca
      601 ttgcactaag tcttgcactt gtcacgaatt cgtacccata cgatgttcca gattacgctg
      661 gatccaaaat tgaagaaggc aaactggtga tttggattaa cggcgataaa ggctataacg
      721 gcctggcgga agtgggcaaa aaatttgaaa aagataccgg cattaaagtg accgtggaac
      781 atccggataa actggaagaa aaatttccgc aggtggcggc gaccggcgat ggcccggata
      841 ttattttttg ggcgcatgat cgctttggcg gctatgcgca gagcggcctg ctggcggaaa
      901 ttaccccgga taaagcgttt caggataaac tgtatccgtt tacctgggat gcggtgcgct
      961 ataacggcaa actgattgcg tatccgattg cggtggaagc gctgagcctg atttataaca
     1021 aagatctgct gccgaacccg ccgaaaacct gggaagaaat tccggcgctg gataaagaac
     1081 tgaaagcgaa aggcaaaagc gcgctgatgt ttaacctgca ggaaccgtat tttacctggc
     1141 cgctgattgc ggcggatggc ggctatgcgt ttaaatatga aaacggcaaa tatgatatta
     1201 aagatgtggg cgtggataac gcgggcgcga aagcgggcct gacctttctg gtggatctga
     1261 ttaaaaacaa acatatgaac gcggataccg attatagcat tgcggaagcg gcgtttaaca
     1321 aaggcgaaac cgcgatgacc attaacggcc cgtgggcgtg gagcaacatt gataccagca
     1381 aagtgaacta tggcgtgacc gtgctgccga cctttaaagg ccagccgagc aaaccgtttg
     1441 tgggcgtgct gagcgcgggc attaacgcgg cgagcccgaa caaagaactg gcgaaagaat
     1501 ttctggaaaa ctatctgctg accgatgaag gcctggaagc ggtgaacaaa gataaaccgc
     1561 tgggcgcggt ggcgctgaaa agctatgaag aagaactggc gaaagatccg cgcattgcgg
     1621 cgaccatgga aaacgcgcag aaaggcgaaa ttatgccgaa cattccgcag atgagcgcgt
     1681 tttggtatgc ggtgcgcacc gcggtgatta acgcggcgag cggccgccag accgtggatg
     1741 aagcgctgaa agatgcgcag accaacgaga acctttactt ccaagctctg gcccccaagg
     1801 aggatccggt gctgaccgtg atcacacagg tgggtctgac catctctctc ctttgcctct
     1861 ttctcgccat tctcaccttc ctgctctgcc gtcccatcca gaacacatcc acgtctctgc
     1921 atctggaact gtctctgtgt ctgtttctgg ctcacttgct gtttctgacc ggcattaaca
     1981 gaacggagcc cgaggtgctc tgtagcatta tcgctggcct gcttcacttc ctctatttgg
     2041 cctgtttcac ttggatgctc ctggaaggac tgcatctctt cctgaccgtc cgcaacctga
     2101 aggttgccaa ctatacgtcc accggcaggt tcaaaaagag gttcatgtac cccgtgggct
     2161 atggcatccc agccgtcatt atcgctgtta gtgcaatcgt aggccctcag aactacggca
     2221 cttttacctg ttggctgaag ctggataagg gcttcatctg gtccttcatg gggcccgtcg
     2281 ccgtgattat cctgatcaac ttggtattct atttccaggt tctgtggatc ctgcggagta
     2341 aactctcatc cctgaacaag gaagtatcaa caatccagga cacccgcgtc atgactttca
     2401 aggcgatctc ccagttgttc atcctggggt gtagctgggg gctggggttt ttcatggtcg
     2461 aagaggtggg gaagaccatc gggtccatta tcgcctatag cttcacaatt atcaatactc
     2521 tgcagggtgt gcttctcttc gttgtgcact gcctgttgaa ccgccaggtg cgcctcatta
     2581 tcctgtccgt gatttctctc gtacccaaat ccaacggttc tgactacaaa gacgatgacg
     2641 acaagtgaag atctgacaaa actcacacat gcccaccgtg cccagcacct gaactcctgg
     2701 ggggaccgtc agtcttcctc ttccccccaa aacccaagga caccctcatg atctcccgga
     2761 cccctgaggt cacatgcgtg gtggtggacg tgagccacga agaccctgag gtcaagttca
     2821 actggtacgt ggacggcgtg gaggtgcata atgccaagac aaagccgcgg gaggagcagt
     2881 acaacagcac gtaccgtgtg gtcagcgtcc tcaccgtcct gcaccaggac tggctgaatg
     2941 gcaaggagta caagtgcaag gtctccaaca aagccctccc agcccccatc gagaaaacca
     3001 tctccaaagc caaagggcag ccccgagaac cacaggtgta caccctgccc ccatcccggg
     3061 aggagatgac caagaaccag gtcagcctga cctgcctggt caaaggcttc tatcccagcg
     3121 acatcgccgt ggagtgggag agcaatgggc agccggagaa caactacaag accacgcctc
     3181 ccgtgctgga ctccgacggc tccttcttcc tctacagcaa gctcaccgtg gacaagagca
     3241 ggtggcagca ggggaacgtc ttctcatgct ccgtgatgca cgaggctctg cacaaccact
     3301 acacgcagaa gagcctctcc ctgtctccgg gtaaatgagt gctagctggc cagacatgat
     3361 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     3421 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     3481 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt
     3541 ttaaagcaag taaaacctct acaaatgtgg tatggaatta attctaaaat acagcatagc
     3601 aaaactttaa cctccaaatc aagcctctac ttgaatcctt ttctgaggga tgaataaggc
     3661 ataggcatca ggggctgttg ccaatgtgca ttagctgttt gcagcctcac cttctttcat
     3721 ggagtttaag atatagtgta ttttcccaag gtttgaacta gctcttcatt tctttatgtt
     3781 ttaaatgcac tgacctccca cattcccttt ttagtaaaat attcagaaat aatttaaata
     3841 catcattgca atgaaaataa atgtttttta ttaggcagaa tccagatgct caaggccctt
     3901 cataatatcc cccagtttag tagttggact tagggaacaa aggaaccttt aatagaaatt
     3961 ggacagcaag aaagcgagct tctagcttat cctcagtcct gctcctctgc cacaaagtgc
     4021 acgcagttgc cggccgggtc gcgcagggcg aactcccgcc cccacggctg ctcgccgatc
     4081 tcggtcatgg ccggcccgga ggcgtcccgg aagttcgtgg acacgacctc cgaccactcg
     4141 gcgtacagct cgtccaggcc gcgcacccac acccaggcca gggtgttgtc cggcaccacc
     4201 tggtcctgga ccgcgctgat gaacagggtc acgtcgtccc ggaccacacc ggcgaagtcg
     4261 tcctccacga agtcccggga gaacccgagc cggtcggtcc agaactcgac cgctccggcg
     4321 acgtcgcgcg cggtgagcac cggaacggca ctggtcaact tggccatgat ggctcctcct
     4381 gtcaggagag gaaagagaag aaggttagta caattgctat agtgagttgt attatactat
     4441 gcagatatac tatgccaatg attaattgtc aaactagggc tgcagggttc atagtgccac
     4501 ttttcctgca ctgccccatc tcctgcccac cctttcccag gcatagacag tcagtgactt
     4561 accaaactca caggagggag aaggcagaag cttgagacag acccgcggga ccgccgaact
     4621 gcgaggggac gtggctaggg cggcttcttt tatggtgcgc cggccctcgg aggcagggcg
     4681 ctcggggagg cctagcggcc aatctgcggt ggcaggaggc ggggccgaag gccgtgcctg
     4741 accaatccgg agcacatagg agtctcagcc ccccgcccca aagcaagggg aagtcacgcg
     4801 cctgtagcgc cagcgtgttg tgaaatgggg gcttgggggg gttggggccc tgactagtca
     4861 aaacaaactc ccattgacgt caatggggtg gagacttgga aatccccgtg agtcaaaccg
     4921 ctatccacgc ccattgatgt actgccaaaa ccgcatcatc atggtaatag cgatgactaa
     4981 tacgtagatg tactgccaag taggaaagtc ccataaggtc atgtactggg cataatgcca
     5041 ggcgggccat ttaccgtcat tgacgtcaat agggggcgta cttggcatat gatacacttg
     5101 atgtactgcc aagtgggcag tttaccgtaa atactccacc cattgacgtc aatggaaagt
     5161 ccctattggc gttactatgg gaacatacgt cattattgac gtcaatgggc gggggtcgtt
     5221 gggcggtcag ccaggcgggc catttaccgt aagttatgta acgcctgcag gttaattaag
     5281 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     5341 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     5401 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     5461 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     5521 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     5581 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     5641 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     5701 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     5761 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     5821 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     5881 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     5941 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     6001 gtcatggcta gttaattaac atttaaatca gcggccgcaa taaaatatct ttattttcat
     6061 tacatctgtg tgttggtttt ttgtgtgaat cgtaactaac atacgctctc catcaaaaca
     6121 aaacgaaaca aaacaaacta gcaaaatagg ctgtccccag tgcaagtgca ggtgccagaa
     6181 catttctcta tcgaa
//
