LOCUS       sequence-430281-        6306 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 6306)
  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 6306)
  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-ADGRE3 CTF-FLAG
FEATURES             Location/Qualifiers
     source          1..6306
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    28..149
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(39..58)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     93..112
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     polyA_signal    253..647
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     CDS             complement(661..1035)
                     /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        1628..1931
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(2011..2599)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2091..2110)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    2707..2755
                     /note="synthetic polyadenylation signal"
     misc_feature    2766..2857
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     2806..2825
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        2894..3105
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             3118..3386
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     sig_peptide     3436..3495
                     /codon_start=1
                     /product="human interleukin-2 signal sequence"
                     /label=IL-2 signal sequence
                     /translation="MYRMQLLSCIALSLALVTNS"
     CDS             3496..3522
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4630..4650
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             5596..5619
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
ORIGIN
        1 aaatgagtgc tagctggcca gacatgataa gatacattga tgagtttgga caaaccacaa
       61 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
      121 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
      181 aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta
      241 tggaattaat tctaaaatac agcatagcaa aactttaacc tccaaatcaa gcctctactt
      301 gaatcctttt ctgagggatg aataaggcat aggcatcagg ggctgttgcc aatgtgcatt
      361 agctgtttgc agcctcacct tctttcatgg agtttaagat atagtgtatt ttcccaaggt
      421 ttgaactagc tcttcatttc tttatgtttt aaatgcactg acctcccaca ttcccttttt
      481 agtaaaatat tcagaaataa tttaaataca tcattgcaat gaaaataaat gttttttatt
      541 aggcagaatc cagatgctca aggcccttca taatatcccc cagtttagta gttggactta
      601 gggaacaaag gaacctttaa tagaaattgg acagcaagaa agcgagcttc tagcttatcc
      661 tcagtcctgc tcctctgcca caaagtgcac gcagttgccg gccgggtcgc gcagggcgaa
      721 ctcccgcccc cacggctgct cgccgatctc ggtcatggcc ggcccggagg cgtcccggaa
      781 gttcgtggac acgacctccg accactcggc gtacagctcg tccaggccgc gcacccacac
      841 ccaggccagg gtgttgtccg gcaccacctg gtcctggacc gcgctgatga acagggtcac
      901 gtcgtcccgg accacaccgg cgaagtcgtc ctccacgaag tcccgggaga acccgagccg
      961 gtcggtccag aactcgaccg ctccggcgac gtcgcgcgcg gtgagcaccg gaacggcact
     1021 ggtcaacttg gccatgatgg ctcctcctgt caggagagga aagagaagaa ggttagtaca
     1081 attgctatag tgagttgtat tatactatgc agatatacta tgccaatgat taattgtcaa
     1141 actagggctg cagggttcat agtgccactt ttcctgcact gccccatctc ctgcccaccc
     1201 tttcccaggc atagacagtc agtgacttac caaactcaca ggagggagaa ggcagaagct
     1261 tgagacagac ccgcgggacc gccgaactgc gaggggacgt ggctagggcg gcttctttta
     1321 tggtgcgccg gccctcggag gcagggcgct cggggaggcc tagcggccaa tctgcggtgg
     1381 caggaggcgg ggccgaaggc cgtgcctgac caatccggag cacataggag tctcagcccc
     1441 ccgccccaaa gcaaggggaa gtcacgcgcc tgtagcgcca gcgtgttgtg aaatgggggc
     1501 ttgggggggt tggggccctg actagtcaaa acaaactccc attgacgtca atggggtgga
     1561 gacttggaaa tccccgtgag tcaaaccgct atccacgccc attgatgtac tgccaaaacc
     1621 gcatcatcat ggtaatagcg atgactaata cgtagatgta ctgccaagta ggaaagtccc
     1681 ataaggtcat gtactgggca taatgccagg cgggccattt accgtcattg acgtcaatag
     1741 ggggcgtact tggcatatga tacacttgat gtactgccaa gtgggcagtt taccgtaaat
     1801 actccaccca ttgacgtcaa tggaaagtcc ctattggcgt tactatggga acatacgtca
     1861 ttattgacgt caatgggcgg gggtcgttgg gcggtcagcc aggcgggcca tttaccgtaa
     1921 gttatgtaac gcctgcaggt taattaagaa catgtgagca aaaggccagc aaaaggccag
     1981 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     2041 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     2101 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     2161 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     2221 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     2281 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     2341 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     2401 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     2461 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     2521 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     2581 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     2641 gaacgaaaac tcacgttaag ggattttggt catggctagt taattaacat ttaaatcagc
     2701 ggccgcaata aaatatcttt attttcatta catctgtgtg ttggtttttt gtgtgaatcg
     2761 taactaacat acgctctcca tcaaaacaaa acgaaacaaa acaaactagc aaaataggct
     2821 gtccccagtg caagtgcagg tgccagaaca tttctctatc gaaggatctg cgatcgctcc
     2881 ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg
     2941 gtcggcaatt gaacgggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc
     3001 gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc
     3061 gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacagctgaa gcttcgaggg
     3121 gctcgcatct ctccttcacg cgcccgccgc cctacctgag gccgccatcc acgccggttg
     3181 agtcgcgttc tgccgcctcc cgcctgtggt gcctcctgaa ctgcgtccgc cgtctaggta
     3241 agtttaaagc tcaggtcgag accgggcctt tgtccggcgc tcccttggag cctacctaga
     3301 ctcagccggc tctccacgct ttgcctgacc ctgcttgctc aactctacgt ctttgtttcg
     3361 ttttctgttc tgcgccgtta cagatccaag ctgtgaccgg cgcctacctg agatcaccgg
     3421 cgaaggaggg ccaccatgta caggatgcaa ctcctgtctt gcattgcact aagtcttgca
     3481 cttgtcacga attcgtaccc atacgatgtt ccagattacg ctggatccaa aattgaagaa
     3541 ggcaaactgg tgatttggat taacggcgat aaaggctata acggcctggc ggaagtgggc
     3601 aaaaaatttg aaaaagatac cggcattaaa gtgaccgtgg aacatccgga taaactggaa
     3661 gaaaaatttc cgcaggtggc ggcgaccggc gatggcccgg atattatttt ttgggcgcat
     3721 gatcgctttg gcggctatgc gcagagcggc ctgctggcgg aaattacccc ggataaagcg
     3781 tttcaggata aactgtatcc gtttacctgg gatgcggtgc gctataacgg caaactgatt
     3841 gcgtatccga ttgcggtgga agcgctgagc ctgatttata acaaagatct gctgccgaac
     3901 ccgccgaaaa cctgggaaga aattccggcg ctggataaag aactgaaagc gaaaggcaaa
     3961 agcgcgctga tgtttaacct gcaggaaccg tattttacct ggccgctgat tgcggcggat
     4021 ggcggctatg cgtttaaata tgaaaacggc aaatatgata ttaaagatgt gggcgtggat
     4081 aacgcgggcg cgaaagcggg cctgaccttt ctggtggatc tgattaaaaa caaacatatg
     4141 aacgcggata ccgattatag cattgcggaa gcggcgttta acaaaggcga aaccgcgatg
     4201 accattaacg gcccgtgggc gtggagcaac attgatacca gcaaagtgaa ctatggcgtg
     4261 accgtgctgc cgacctttaa aggccagccg agcaaaccgt ttgtgggcgt gctgagcgcg
     4321 ggcattaacg cggcgagccc gaacaaagaa ctggcgaaag aatttctgga aaactatctg
     4381 ctgaccgatg aaggcctgga agcggtgaac aaagataaac cgctgggcgc ggtggcgctg
     4441 aaaagctatg aagaagaact ggcgaaagat ccgcgcattg cggcgaccat ggaaaacgcg
     4501 cagaaaggcg aaattatgcc gaacattccg cagatgagcg cgttttggta tgcggtgcgc
     4561 accgcggtga ttaacgcggc gagcggccgc cagaccgtgg atgaagcgct gaaagatgcg
     4621 cagaccaacg agaaccttta cttccaatcc agctttgctg tcctgatggc gcttacttct
     4681 caagaagagg accctgtgct gaccgttatc acctacgtag gactctcagt ctccctcttg
     4741 tgccttctcc tggctgccct cactttcctc ctgtgtaagg caattcgcaa cacttctacc
     4801 agcctgcatc tccagctgtc tctgtgcctg ttcctcgccc acctcctgtt cctggttggg
     4861 atcgatcgta ctgaacctaa ggttctgtgt tccattatcg ccggtgctct ccattatctt
     4921 tatctggctg cctttacctg gatgcttctg gagggcgtgc acctgttcct gaccgctagg
     4981 aacctgactg tggttaacta cagctctatc aacagactca tgaagtggat tatgttccct
     5041 gtgggctatg gggtgcctgc cgtgactgtc gccatcagtg ctgcatcatg gccacacctg
     5101 tacggcacag ccgaccggtg ctggctgcac ctggaccagg gatttatgtg gagctttctg
     5161 gggcccgttt gcgctatttt ctctgctaac ctggtgctgt tcattttggt cttctggatc
     5221 ctcaaacgta agcttagctc tctgaactct gaagtgtcca cgatccagaa tacccgcatg
     5281 ctggccttca aagccacagc ccagctgttt atcctgggct gtacctggtg tctgggcttg
     5341 ctccaggtcg gaccggcagc ccaggtgatg gcctacctgt ttactattat caacagcttg
     5401 cagggttttt tcatctttct tgtgtattgc ttgctgtccc aacaggtgca gaagcaatac
     5461 cagaagtggt tccgggagat cgtaaagtcc aagagcgagt ccgaaaccta tacactgtct
     5521 agcaagatgg gccctgactc taaaccatcc gagggagatg tttttcccgg ccaggtaaaa
     5581 cgcaagtacg gttctgacta caaagacgat gacgacaagt gaagatctga caaaactcac
     5641 acatgcccac cgtgcccagc acctgaactc ctggggggac cgtcagtctt cctcttcccc
     5701 ccaaaaccca aggacaccct catgatctcc cggacccctg aggtcacatg cgtggtggtg
     5761 gacgtgagcc acgaagaccc tgaggtcaag ttcaactggt acgtggacgg cgtggaggtg
     5821 cataatgcca agacaaagcc gcgggaggag cagtacaaca gcacgtaccg tgtggtcagc
     5881 gtcctcaccg tcctgcacca ggactggctg aatggcaagg agtacaagtg caaggtctcc
     5941 aacaaagccc tcccagcccc catcgagaaa accatctcca aagccaaagg gcagccccga
     6001 gaaccacagg tgtacaccct gcccccatcc cgggaggaga tgaccaagaa ccaggtcagc
     6061 ctgacctgcc tggtcaaagg cttctatccc agcgacatcg ccgtggagtg ggagagcaat
     6121 gggcagccgg agaacaacta caagaccacg cctcccgtgc tggactccga cggctccttc
     6181 ttcctctaca gcaagctcac cgtggacaag agcaggtggc agcaggggaa cgtcttctca
     6241 tgctccgtga tgcacgaggc tctgcacaac cactacacgc agaagagcct ctccctgtct
     6301 ccgggt
//
