LOCUS       sequence-430131-        7116 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 7116)
  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 7116)
  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-ADGRA3 CTF-FLAG
FEATURES             Location/Qualifiers
     source          1..7116
                     /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             3561..3584
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    4299..4420
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4310..4329)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4364..4383
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     polyA_signal    4524..4918
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     CDS             complement(4932..5306)
                     /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        5899..6202
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(6282..6870)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6362..6381)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    6978..7026
                     /note="synthetic polyadenylation signal"
     misc_feature    join(7037..7116,1..12)
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     7077..7096
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
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 aatccaacta cgccgtcctc atggatctga ccgggtccga gctgtacact caagcggctt
     1861 ctctgcttca ccccgttgtg tacaccactg ctattatcct gttgctgtgc ttgctggctg
     1921 ttatcgttag ctacatttat catcacagcc tcatccgtat ctctcttaag tcatggcata
     1981 tgctggttaa tctgtgtttc cacatctttc tgacctgcgt tgtgttcgtt gggggcatta
     2041 cacagaccag gaacgcttct atctgccagg cggttggcat tatcctccat tactccaccc
     2101 tggctactgt gctctgggta ggggtcaccg cccgcaatat ctacaagcag gtcacaaaga
     2161 aagctaagcg ctgccaggat ccggatgaac cccctccccc tccccgtcca atgttgcgtt
     2221 tttacctgat cgggggcggt atccctatta tcgtgtgcgg catcaccgca gccgctaaca
     2281 tcaagaatta cggctcccgc ccgaacgccc catactgctg gatggcgtgg gaaccctccc
     2341 ttggcgcttt ctatgggcca gcttctttca tcacattcgt caactgtatg tactttctgt
     2401 ccattttcat tcagttgaaa agacaccccg agcgtaagta cgagctcaag gagccgacag
     2461 aagagcaaca gcgcctggca gccaacgaga acggagaaat taaccaccag gacagcatga
     2521 gtctctccct gatcagcaca tctgccctcg agaacgaaca caccttccac tctcagctgt
     2581 tgggcgccag cttgaccctg ctcctgtacg tggcgctgtg gatgttcggc gctctggccg
     2641 tctccctgta ctatccactg gacctggtgt tctcatttgt gtttggtgcc acctccctga
     2701 gtttcagcgc ttttttcgtc gtgcatcact gtgtgaaccg cgaagacgtt cgtttggctt
     2761 ggatcatgac ctgttgcccg ggcaggtcta gctattccgt gcaggtgaat gtccagcccc
     2821 ctaactctaa cggtaccaac ggggaggcgc ccaagtgtcc gaactcttcc gctgagtcta
     2881 gttgcactaa caaatctgct tctagcttca aaaactcatc tcaggggtgc aagttgacaa
     2941 atcttcaggc cgcggctgcc cagtgccatg caaactctct gcctttgaac agcaccccac
     3001 agctggacaa ctccctgact gagcactcca tggacaacga cattaaaatg cacgtggctc
     3061 cattggaggt gcaattccgc actaatgtac actcttccag gcatcacaag aaccggagca
     3121 aaggccatag agcgtctcgc ctgactgtgc tccgcgagta tgcctacgat gtgccgacta
     3181 gtgtcgaggg ttccgtacaa aatggcctgc ccaagagtcg cctcggtaat aacgaggggc
     3241 acagcaggtc tcgtcgggcc tacctcgcat accgtgagcg tcagtacaat ccccctcaac
     3301 aggactcctc tgatgcctgt agtaccctgc caaagtctag cagaaacttc gaaaagccag
     3361 tcagcacaac cagcaaaaag gacgcccttc ggaagcctgc tgtggttgaa ctcgagaacc
     3421 aacagaaaag ttacggtctt aacctcgcga tccaaaacgg tccaatcaag tccaacggac
     3481 aggaggggcc cttgctgggc acagacagca caggcaacgt tcggaccggt ttgtggaagc
     3541 acgaaaccac cgtcggttct gactacaaag acgatgacga caagtgaaga tctgacaaaa
     3601 ctcacacatg cccaccgtgc ccagcacctg aactcctggg gggaccgtca gtcttcctct
     3661 tccccccaaa acccaaggac accctcatga tctcccggac ccctgaggtc acatgcgtgg
     3721 tggtggacgt gagccacgaa gaccctgagg tcaagttcaa ctggtacgtg gacggcgtgg
     3781 aggtgcataa tgccaagaca aagccgcggg aggagcagta caacagcacg taccgtgtgg
     3841 tcagcgtcct caccgtcctg caccaggact ggctgaatgg caaggagtac aagtgcaagg
     3901 tctccaacaa agccctccca gcccccatcg agaaaaccat ctccaaagcc aaagggcagc
     3961 cccgagaacc acaggtgtac accctgcccc catcccggga ggagatgacc aagaaccagg
     4021 tcagcctgac ctgcctggtc aaaggcttct atcccagcga catcgccgtg gagtgggaga
     4081 gcaatgggca gccggagaac aactacaaga ccacgcctcc cgtgctggac tccgacggct
     4141 ccttcttcct ctacagcaag ctcaccgtgg acaagagcag gtggcagcag gggaacgtct
     4201 tctcatgctc cgtgatgcac gaggctctgc acaaccacta cacgcagaag agcctctccc
     4261 tgtctccggg taaatgagtg ctagctggcc agacatgata agatacattg atgagtttgg
     4321 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     4381 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     4441 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
     4501 caaatgtggt atggaattaa ttctaaaata cagcatagca aaactttaac ctccaaatca
     4561 agcctctact tgaatccttt tctgagggat gaataaggca taggcatcag gggctgttgc
     4621 caatgtgcat tagctgtttg cagcctcacc ttctttcatg gagtttaaga tatagtgtat
     4681 tttcccaagg tttgaactag ctcttcattt ctttatgttt taaatgcact gacctcccac
     4741 attccctttt tagtaaaata ttcagaaata atttaaatac atcattgcaa tgaaaataaa
     4801 tgttttttat taggcagaat ccagatgctc aaggcccttc ataatatccc ccagtttagt
     4861 agttggactt agggaacaaa ggaaccttta atagaaattg gacagcaaga aagcgagctt
     4921 ctagcttatc ctcagtcctg ctcctctgcc acaaagtgca cgcagttgcc ggccgggtcg
     4981 cgcagggcga actcccgccc ccacggctgc tcgccgatct cggtcatggc cggcccggag
     5041 gcgtcccgga agttcgtgga cacgacctcc gaccactcgg cgtacagctc gtccaggccg
     5101 cgcacccaca cccaggccag ggtgttgtcc ggcaccacct ggtcctggac cgcgctgatg
     5161 aacagggtca cgtcgtcccg gaccacaccg gcgaagtcgt cctccacgaa gtcccgggag
     5221 aacccgagcc ggtcggtcca gaactcgacc gctccggcga cgtcgcgcgc ggtgagcacc
     5281 ggaacggcac tggtcaactt ggccatgatg gctcctcctg tcaggagagg aaagagaaga
     5341 aggttagtac aattgctata gtgagttgta ttatactatg cagatatact atgccaatga
     5401 ttaattgtca aactagggct gcagggttca tagtgccact tttcctgcac tgccccatct
     5461 cctgcccacc ctttcccagg catagacagt cagtgactta ccaaactcac aggagggaga
     5521 aggcagaagc ttgagacaga cccgcgggac cgccgaactg cgaggggacg tggctagggc
     5581 ggcttctttt atggtgcgcc ggccctcgga ggcagggcgc tcggggaggc ctagcggcca
     5641 atctgcggtg gcaggaggcg gggccgaagg ccgtgcctga ccaatccgga gcacatagga
     5701 gtctcagccc cccgccccaa agcaagggga agtcacgcgc ctgtagcgcc agcgtgttgt
     5761 gaaatggggg cttggggggg ttggggccct gactagtcaa aacaaactcc cattgacgtc
     5821 aatggggtgg agacttggaa atccccgtga gtcaaaccgc tatccacgcc cattgatgta
     5881 ctgccaaaac cgcatcatca tggtaatagc gatgactaat acgtagatgt actgccaagt
     5941 aggaaagtcc cataaggtca tgtactgggc ataatgccag gcgggccatt taccgtcatt
     6001 gacgtcaata gggggcgtac ttggcatatg atacacttga tgtactgcca agtgggcagt
     6061 ttaccgtaaa tactccaccc attgacgtca atggaaagtc cctattggcg ttactatggg
     6121 aacatacgtc attattgacg tcaatgggcg ggggtcgttg ggcggtcagc caggcgggcc
     6181 atttaccgta agttatgtaa cgcctgcagg ttaattaaga acatgtgagc aaaaggccag
     6241 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     6301 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     6361 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     6421 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     6481 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     6541 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     6601 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     6661 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     6721 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     6781 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     6841 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     6901 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatggctag ttaattaaca
     6961 tttaaatcag cggccgcaat aaaatatctt tattttcatt acatctgtgt gttggttttt
     7021 tgtgtgaatc gtaactaaca tacgctctcc atcaaaacaa aacgaaacaa aacaaactag
     7081 caaaataggc tgtccccagt gcaagtgcag gtgcca
//
