LOCUS       sequence-430006-        7140 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 7140)
  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 7140)
  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-ADGRA2 CTF-FLAG
FEATURES             Location/Qualifiers
     source          1..7140
                     /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="ENLYFQG"
     CDS             3567..3590
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    4305..4426
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4316..4335)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4370..4389
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     polyA_signal    4530..4924
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     CDS             complement(4938..5312)
                     /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        5905..6208
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(6288..6876)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6368..6387)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    6984..7032
                     /note="synthetic polyadenylation signal"
     misc_feature    7043..7134
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     7083..7102
                     /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 ccaaggcaat gtggcggtgc
     1801 tgatggaact gtctgccttt cctcgcgagg tcggtggcgc tggagcggga ctccaccctg
     1861 ttgtgtatcc ctgtacagcc ttgctgttgc tgtgcttgtt cgccactatc attacttaca
     1921 ttctgaatca ctcctcaatt cgggtatccc gcaagggatg gcacatgctc ctgaacctct
     1981 gtttccacat cgccatgaca agcgccgtct tcgccggtgg catcaccctg accaattatc
     2041 agatggtgtg ccaggccgta ggtatcaccc tgcactattc ctctctctcc accctcctgt
     2101 ggatgggcgt aaaggcccgt gtgcttcaca aggaactgac gtggagggct cctccaccgc
     2161 aggagggaga tcccgccctg cctacaccaa gtcccatgct gcgcttctac ctgatcgccg
     2221 gcggtatccc attgatcatt tgcggcatca cggctgcggt caatatccac aattacagag
     2281 atcatagtcc ctattgctgg ctcgtgtggc gtccttcact gggagctttc tacattcccg
     2341 ttgctctcat ccttctgatt acttggattt acttcctgtg tgccgggttg cgcctccgtg
     2401 ggcctctcgc gcagaatcct aaggccggga actccagggc ctccctggag gccggagagg
     2461 aactgcgtgg ctccacacgt ctgagaggaa gcggcccact gttgtctgat tccggctccc
     2521 ttctggccac tggatctgct cgtgtgggaa ctcccggtcc gccagaagat ggtgactcct
     2581 tgtacagtcc cggcgtccag ctgggagcct tggtcacaac ccattttctg tatctggcta
     2641 tgtgggcctg tggtgccctg gcggttagcc agcgttggct gcctcgcgtt gtgtgcagct
     2701 gtctctacgg agtcgcagcc tccgctttgg gcctcttcgt ttttactcat cactgtgcta
     2761 gacggcgcga cgtgagagca tcatggcgtg cctgttgccc tccggcatca ccagccgctc
     2821 cgcacgcacc acctcgcgct ctgccggcag ctgccgaaga tggttctcct gtctttggcg
     2881 agggcccgcc cagcctcaag tctagcccta gtggaagctc cggccacccg ctggcactgg
     2941 gaccgtgcaa gttgaccaac ctgcagttgg ctcagagcca agtgtgcgaa gccggtgctg
     3001 cggctggcgg tgagggcgaa ccagaacccg ccggtacaag aggcaatctg gcccataggc
     3061 acccgaacaa tgttcatcac ggtaggcgtg ctcataagtc tcgtgccaaa gggcaccgtg
     3121 cgggtgaagc ctgtggtaag aaccgcctca aagctttgcg cggtggcgcg gccggtgctc
     3181 tggagcttct gagcagtgaa tctggctctc tgcacaacag tcccactgac tcttatctcg
     3241 ggtcctcacg caactcacca ggagccggtc tccaactgga gggcgaacca atgctgactc
     3301 catccgaagg tagcgacacc tcagcggcac cgttgtccga agccggaagg gcagggcaac
     3361 gtcgcagcgc ctcacgtgac agcctcaagg gcggaggcgc cctggaaaag gaaagtcata
     3421 ggcgcagcta cccgctgaac gctgccagcc tgaatggtgc tcccaaggga ggtaagtatg
     3481 atgacgtgac ccttatgggc gccgaggttg ccagtggcgg ttgcatgaag accggtctgt
     3541 ggaaatcaga gactaccgtc ggttctgact acaaagacga tgacgacaag tgaagatctg
     3601 acaaaactca cacatgccca ccgtgcccag cacctgaact cctgggggga ccgtcagtct
     3661 tcctcttccc cccaaaaccc aaggacaccc tcatgatctc ccggacccct gaggtcacat
     3721 gcgtggtggt ggacgtgagc cacgaagacc ctgaggtcaa gttcaactgg tacgtggacg
     3781 gcgtggaggt gcataatgcc aagacaaagc cgcgggagga gcagtacaac agcacgtacc
     3841 gtgtggtcag cgtcctcacc gtcctgcacc aggactggct gaatggcaag gagtacaagt
     3901 gcaaggtctc caacaaagcc ctcccagccc ccatcgagaa aaccatctcc aaagccaaag
     3961 ggcagccccg agaaccacag gtgtacaccc tgcccccatc ccgggaggag atgaccaaga
     4021 accaggtcag cctgacctgc ctggtcaaag gcttctatcc cagcgacatc gccgtggagt
     4081 gggagagcaa tgggcagccg gagaacaact acaagaccac gcctcccgtg ctggactccg
     4141 acggctcctt cttcctctac agcaagctca ccgtggacaa gagcaggtgg cagcagggga
     4201 acgtcttctc atgctccgtg atgcacgagg ctctgcacaa ccactacacg cagaagagcc
     4261 tctccctgtc tccgggtaaa tgagtgctag ctggccagac atgataagat acattgatga
     4321 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     4381 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     4441 cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa
     4501 cctctacaaa tgtggtatgg aattaattct aaaatacagc atagcaaaac tttaacctcc
     4561 aaatcaagcc tctacttgaa tccttttctg agggatgaat aaggcatagg catcaggggc
     4621 tgttgccaat gtgcattagc tgtttgcagc ctcaccttct ttcatggagt ttaagatata
     4681 gtgtattttc ccaaggtttg aactagctct tcatttcttt atgttttaaa tgcactgacc
     4741 tcccacattc cctttttagt aaaatattca gaaataattt aaatacatca ttgcaatgaa
     4801 aataaatgtt ttttattagg cagaatccag atgctcaagg cccttcataa tatcccccag
     4861 tttagtagtt ggacttaggg aacaaaggaa cctttaatag aaattggaca gcaagaaagc
     4921 gagcttctag cttatcctca gtcctgctcc tctgccacaa agtgcacgca gttgccggcc
     4981 gggtcgcgca gggcgaactc ccgcccccac ggctgctcgc cgatctcggt catggccggc
     5041 ccggaggcgt cccggaagtt cgtggacacg acctccgacc actcggcgta cagctcgtcc
     5101 aggccgcgca cccacaccca ggccagggtg ttgtccggca ccacctggtc ctggaccgcg
     5161 ctgatgaaca gggtcacgtc gtcccggacc acaccggcga agtcgtcctc cacgaagtcc
     5221 cgggagaacc cgagccggtc ggtccagaac tcgaccgctc cggcgacgtc gcgcgcggtg
     5281 agcaccggaa cggcactggt caacttggcc atgatggctc ctcctgtcag gagaggaaag
     5341 agaagaaggt tagtacaatt gctatagtga gttgtattat actatgcaga tatactatgc
     5401 caatgattaa ttgtcaaact agggctgcag ggttcatagt gccacttttc ctgcactgcc
     5461 ccatctcctg cccacccttt cccaggcata gacagtcagt gacttaccaa actcacagga
     5521 gggagaaggc agaagcttga gacagacccg cgggaccgcc gaactgcgag gggacgtggc
     5581 tagggcggct tcttttatgg tgcgccggcc ctcggaggca gggcgctcgg ggaggcctag
     5641 cggccaatct gcggtggcag gaggcggggc cgaaggccgt gcctgaccaa tccggagcac
     5701 ataggagtct cagccccccg ccccaaagca aggggaagtc acgcgcctgt agcgccagcg
     5761 tgttgtgaaa tgggggcttg ggggggttgg ggccctgact agtcaaaaca aactcccatt
     5821 gacgtcaatg gggtggagac ttggaaatcc ccgtgagtca aaccgctatc cacgcccatt
     5881 gatgtactgc caaaaccgca tcatcatggt aatagcgatg actaatacgt agatgtactg
     5941 ccaagtagga aagtcccata aggtcatgta ctgggcataa tgccaggcgg gccatttacc
     6001 gtcattgacg tcaatagggg gcgtacttgg catatgatac acttgatgta ctgccaagtg
     6061 ggcagtttac cgtaaatact ccacccattg acgtcaatgg aaagtcccta ttggcgttac
     6121 tatgggaaca tacgtcatta ttgacgtcaa tgggcggggg tcgttgggcg gtcagccagg
     6181 cgggccattt accgtaagtt atgtaacgcc tgcaggttaa ttaagaacat gtgagcaaaa
     6241 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     6301 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     6361 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     6421 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     6481 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     6541 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     6601 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     6661 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     6721 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     6781 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     6841 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     6901 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat ggctagttaa
     6961 ttaacattta aatcagcggc cgcaataaaa tatctttatt ttcattacat ctgtgtgttg
     7021 gttttttgtg tgaatcgtaa ctaacatacg ctctccatca aaacaaaacg aaacaaaaca
     7081 aactagcaaa ataggctgtc cccagtgcaa gtgcaggtgc cagaacattt ctctatcgaa
//
