LOCUS       sequence-429999-        7044 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 7044)
  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 7044)
  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-ADGRA1-FLAG
FEATURES             Location/Qualifiers
     source          1..7044
                     /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="ENLYFQM"
     CDS             3471..3494
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    4209..4330
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4220..4239)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4274..4293
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     polyA_signal    4434..4828
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     CDS             complement(4842..5216)
                     /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        5809..6112
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(6192..6780)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6272..6291)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    6888..6936
                     /note="synthetic polyadenylation signal"
     misc_feature    6947..7038
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     6987..7006
                     /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 ccaaatggac ctgaagactg
     1801 tactctcctt gccacgctat cccggcgagt ttcttcatcc cgtcgtgtac gcctgcaccg
     1861 ccgttatgct tctgtgcctc ctggcttctt tcgtgaccta cattgtccat cagtctgcga
     1921 tcagaatttc ccgcaaaggg cgccataccc tcctgaactt ctgcttccac gcagctctga
     1981 cctttacggt gttcgccgga ggcatcaacc gtactaagta cccaatcctc tgtcaagcag
     2041 ttgggattgt cctgcactac agcaccctta gtaccatgct ttggattggc gtaaccgccc
     2101 gtaacattta caagcaggtc accaaaaagg cgccactgtg cctggacaca gatcaacctc
     2161 cctacccaag acagcctctg cttagattct acctcgtgag cggaggcgtg ccgtttatca
     2221 tttgtggagt cactgccgca accaacatcc gcaactacgg tacagaggat gaggatactg
     2281 cttactgttg gatggcatgg gaaccatctc tgggcgcctt ctacggtccc gctgccatta
     2341 tcacactggt aacgtgtgtg tacttcctgg gcacttacgt gcaactgcgc agacatccgg
     2401 gcaggcgcta cgagctgaga actcagcccg aagagcagcg ccggttggct actccagagg
     2461 gtggcagagg tatccgccca ggcacacctc cagcccacga cgctccaggt gcaagtgtgc
     2521 ttcagaacga acactcattc caggcccaac tgagggcagc ggcctttact ctgttcctct
     2581 tcaccgctac ctgggcgttt ggagccctgg ctgtgtccca agggcacttc ctggacatgg
     2641 tgttctcctg tctgtacggc gcgttctgcg ttaccctcgg actcttcgtg ctgattcatc
     2701 actgcgccaa acgcgaagac gtttggcagt gctggtgggc atgctgtcca cctcggaaag
     2761 acgcgcatcc tgccctggac gctaacggag ccgctcttgg tcgggcagct tgcctgcaca
     2821 gccctggact cgggcagcct cgcggcttcg ctcatccacc cggtccgtgt aagatgacca
     2881 acctccaggc ggctcagggc cacgcctcct gcctgagccc tgctacaccc tgttgcgcga
     2941 agatgcactg cgagccgctg accgcagacg aggctcacgt gcatctccag gaagagggcg
     3001 ccttcggcca cgatccgcat ctgcatgggt gcttgcaggg ccgtactaag ccgccctact
     3061 tctcccgcca tcccgccgag gaacccgaat acgcatacca cattcccagt tccttggacg
     3121 gcagtccaag atccagccgc acggactcac caccgagctc tctggacggc ccagccggta
     3181 ctcacacctt ggcctgttgc acccagggag acccttttcc catggtgacc cagcctgagg
     3241 gctctgacgg ttcaccggca ctgtattcct gcccaaccca gcccggtagg gaggcagccc
     3301 tcgggccagg ccacctggaa atgctgcgca ggactcagag tcttcccttc ggcggtccct
     3361 cccagaatgg tctgccaaaa gggaagttgc tggaaggcct gccatttggc accgatggca
     3421 ccggaaacat tcgcacggga ccttggaaga acgagacgac tgtgggttct gactacaaag
     3481 acgatgacga caagtgaaga tctgacaaaa ctcacacatg cccaccgtgc ccagcacctg
     3541 aactcctggg gggaccgtca gtcttcctct tccccccaaa acccaaggac accctcatga
     3601 tctcccggac ccctgaggtc acatgcgtgg tggtggacgt gagccacgaa gaccctgagg
     3661 tcaagttcaa ctggtacgtg gacggcgtgg aggtgcataa tgccaagaca aagccgcggg
     3721 aggagcagta caacagcacg taccgtgtgg tcagcgtcct caccgtcctg caccaggact
     3781 ggctgaatgg caaggagtac aagtgcaagg tctccaacaa agccctccca gcccccatcg
     3841 agaaaaccat ctccaaagcc aaagggcagc cccgagaacc acaggtgtac accctgcccc
     3901 catcccggga ggagatgacc aagaaccagg tcagcctgac ctgcctggtc aaaggcttct
     3961 atcccagcga catcgccgtg gagtgggaga gcaatgggca gccggagaac aactacaaga
     4021 ccacgcctcc cgtgctggac tccgacggct ccttcttcct ctacagcaag ctcaccgtgg
     4081 acaagagcag gtggcagcag gggaacgtct tctcatgctc cgtgatgcac gaggctctgc
     4141 acaaccacta cacgcagaag agcctctccc tgtctccggg taaatgagtg ctagctggcc
     4201 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     4261 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     4321 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg
     4381 ggaggttttt taaagcaagt aaaacctcta caaatgtggt atggaattaa ttctaaaata
     4441 cagcatagca aaactttaac ctccaaatca agcctctact tgaatccttt tctgagggat
     4501 gaataaggca taggcatcag gggctgttgc caatgtgcat tagctgtttg cagcctcacc
     4561 ttctttcatg gagtttaaga tatagtgtat tttcccaagg tttgaactag ctcttcattt
     4621 ctttatgttt taaatgcact gacctcccac attccctttt tagtaaaata ttcagaaata
     4681 atttaaatac atcattgcaa tgaaaataaa tgttttttat taggcagaat ccagatgctc
     4741 aaggcccttc ataatatccc ccagtttagt agttggactt agggaacaaa ggaaccttta
     4801 atagaaattg gacagcaaga aagcgagctt ctagcttatc ctcagtcctg ctcctctgcc
     4861 acaaagtgca cgcagttgcc ggccgggtcg cgcagggcga actcccgccc ccacggctgc
     4921 tcgccgatct cggtcatggc cggcccggag gcgtcccgga agttcgtgga cacgacctcc
     4981 gaccactcgg cgtacagctc gtccaggccg cgcacccaca cccaggccag ggtgttgtcc
     5041 ggcaccacct ggtcctggac cgcgctgatg aacagggtca cgtcgtcccg gaccacaccg
     5101 gcgaagtcgt cctccacgaa gtcccgggag aacccgagcc ggtcggtcca gaactcgacc
     5161 gctccggcga cgtcgcgcgc ggtgagcacc ggaacggcac tggtcaactt ggccatgatg
     5221 gctcctcctg tcaggagagg aaagagaaga aggttagtac aattgctata gtgagttgta
     5281 ttatactatg cagatatact atgccaatga ttaattgtca aactagggct gcagggttca
     5341 tagtgccact tttcctgcac tgccccatct cctgcccacc ctttcccagg catagacagt
     5401 cagtgactta ccaaactcac aggagggaga aggcagaagc ttgagacaga cccgcgggac
     5461 cgccgaactg cgaggggacg tggctagggc ggcttctttt atggtgcgcc ggccctcgga
     5521 ggcagggcgc tcggggaggc ctagcggcca atctgcggtg gcaggaggcg gggccgaagg
     5581 ccgtgcctga ccaatccgga gcacatagga gtctcagccc cccgccccaa agcaagggga
     5641 agtcacgcgc ctgtagcgcc agcgtgttgt gaaatggggg cttggggggg ttggggccct
     5701 gactagtcaa aacaaactcc cattgacgtc aatggggtgg agacttggaa atccccgtga
     5761 gtcaaaccgc tatccacgcc cattgatgta ctgccaaaac cgcatcatca tggtaatagc
     5821 gatgactaat acgtagatgt actgccaagt aggaaagtcc cataaggtca tgtactgggc
     5881 ataatgccag gcgggccatt taccgtcatt gacgtcaata gggggcgtac ttggcatatg
     5941 atacacttga tgtactgcca agtgggcagt ttaccgtaaa tactccaccc attgacgtca
     6001 atggaaagtc cctattggcg ttactatggg aacatacgtc attattgacg tcaatgggcg
     6061 ggggtcgttg ggcggtcagc caggcgggcc atttaccgta agttatgtaa cgcctgcagg
     6121 ttaattaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     6181 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     6241 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     6301 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     6361 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     6421 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     6481 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     6541 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     6601 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     6661 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     6721 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     6781 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     6841 gggattttgg tcatggctag ttaattaaca tttaaatcag cggccgcaat aaaatatctt
     6901 tattttcatt acatctgtgt gttggttttt tgtgtgaatc gtaactaaca tacgctctcc
     6961 atcaaaacaa aacgaaacaa aacaaactag caaaataggc tgtccccagt gcaagtgcag
     7021 gtgccagaac atttctctat cgaa
//
