LOCUS       sequence-430136-        7362 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 7362)
  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 7362)
  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-ADGRB3 CTF-FLAG
FEATURES             Location/Qualifiers
     source          1..7362
                     /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             3807..3830
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    4545..4666
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4556..4575)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4610..4629
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     polyA_signal    4770..5164
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     CDS             complement(5178..5552)
                     /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        6145..6448
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(6528..7116)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6608..6627)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    7224..7272
                     /note="synthetic polyadenylation signal"
     misc_feature    join(7283..7362,1..12)
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     7323..7342
                     /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 aatctacctt cgccattttg gctcagcaac ctagagaaat aatcatggaa tcctctggca
     1861 caccttcagt taccctaata gtaggcagtg gtctttcttg cttggccttg attaccctag
     1921 cagttgtcta tgcagcatta tggaggtaca tacgctctga gagatccata atactaatta
     1981 acttctgcct gtctatcatc tcatccaata tcctcatact ggttggacag actcagacac
     2041 ataataagag tatctgcaca accaccactg catttttgca ctttttcttc ctggcttcat
     2101 tctgttgggt tttgactgag gcgtggcaat catatatggc tgtaactgga aaaattagga
     2161 cacggcttat aagaaaacgc tttttgtgcc ttggatgggg tttaccagca ttagtagtgg
     2221 ccacatcagt aggcttcacc agaacaaaag gatatggcac tgatcactac tgctggctct
     2281 ctcttgaagg aggactactc tatgcttttg tgggacctgc agccgctgtt gtcctggtca
     2341 acatggtgat tggcattttg gtatttaata aacttgtttc cagagatgga atcctagata
     2401 aaaagctcaa acacagagcc ggtcagatga gtgagcctca tagcggtttg acgctcaaat
     2461 gtgccaagtg tggagtagtt tcaacaacag ctttgtcagc caccaccgcc agtaacgcca
     2521 tggcgtctct ttggagctcc tgtgtggtgt tgccccttct ggctttgacg tggatgtctg
     2581 cggttctggc catgacagat aaacgctcca tattgtttca aatacttttt gctgtgtttg
     2641 attcattgca aggctttgtt atagtcatgg tccactgcat tcttcggaga gaggttcagg
     2701 atgcatttag atgccgattg agaaactgtc aggatcccat caatgcagat tcttcgagtt
     2761 cgtttcctaa tgggcatgct caaatcatga cagactttga aaaggatgta gacattgcct
     2821 gtcgatcagt tcttcataag gatattggtc cttgccgagc agccacaata acaggaacac
     2881 tttctaggat ttctctaaat gatgatgaag aagaaaaggg aacaaaccct gaagggctaa
     2941 gctattcaac attgcctgga aatgtcattt ccaaagtcat catccagcaa cccacaggtt
     3001 tgcacatgcc catgagtatg aatgagctta gcaatccatg tttgaaaaaa gaaaatagtg
     3061 aattgcggag aactgtgtac ttatgtacgg atgataattt gagaggggct gacatggaca
     3121 tagtccatcc tcaagaaaga atgatggaaa gtgactatat tgtgatgccc agaagttctg
     3181 taaataacca gccttcaatg aaagaagaaa gcaaaatgaa tattggcatg gaaaccttgc
     3241 cgcatgaaag gctattgcac tacaaagtaa accctgaatt caatatgaat ccccctgtaa
     3301 tggaccagtt caatatgaac ttagagcaac atctcgcacc ccaggaacat atgcagaatt
     3361 tgccctttga acctcgcaca gctgtgaaga atttcatggc ctctgagttg gatgataatg
     3421 caggactatc aagaagtgaa actggatcaa cgatatcaat gagttcttta gagagaagaa
     3481 aatcacgata ttcagacctt gactttgaga aggtcatgca tacaaggaag aggcatatgg
     3541 aactatttca agaactaaat cagaaatttc aaactttgga cagatttcgg gatataccaa
     3601 atacaagcag tatggaaaac cccgcaccaa acaagaatcc atgggacact ttcaaaaacc
     3661 ccagtgaata cccgcattac accacaatca atgtcttaga cacagaggca aaggatgctt
     3721 tggaactgag gccagcagag tgggagaagt gtctgaattt gcctctggat gtgcaagagg
     3781 gtgactttca aacagaagtt ggttctgact acaaagacga tgacgacaag tgaagatctg
     3841 acaaaactca cacatgccca ccgtgcccag cacctgaact cctgggggga ccgtcagtct
     3901 tcctcttccc cccaaaaccc aaggacaccc tcatgatctc ccggacccct gaggtcacat
     3961 gcgtggtggt ggacgtgagc cacgaagacc ctgaggtcaa gttcaactgg tacgtggacg
     4021 gcgtggaggt gcataatgcc aagacaaagc cgcgggagga gcagtacaac agcacgtacc
     4081 gtgtggtcag cgtcctcacc gtcctgcacc aggactggct gaatggcaag gagtacaagt
     4141 gcaaggtctc caacaaagcc ctcccagccc ccatcgagaa aaccatctcc aaagccaaag
     4201 ggcagccccg agaaccacag gtgtacaccc tgcccccatc ccgggaggag atgaccaaga
     4261 accaggtcag cctgacctgc ctggtcaaag gcttctatcc cagcgacatc gccgtggagt
     4321 gggagagcaa tgggcagccg gagaacaact acaagaccac gcctcccgtg ctggactccg
     4381 acggctcctt cttcctctac agcaagctca ccgtggacaa gagcaggtgg cagcagggga
     4441 acgtcttctc atgctccgtg atgcacgagg ctctgcacaa ccactacacg cagaagagcc
     4501 tctccctgtc tccgggtaaa tgagtgctag ctggccagac atgataagat acattgatga
     4561 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     4621 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     4681 cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa
     4741 cctctacaaa tgtggtatgg aattaattct aaaatacagc atagcaaaac tttaacctcc
     4801 aaatcaagcc tctacttgaa tccttttctg agggatgaat aaggcatagg catcaggggc
     4861 tgttgccaat gtgcattagc tgtttgcagc ctcaccttct ttcatggagt ttaagatata
     4921 gtgtattttc ccaaggtttg aactagctct tcatttcttt atgttttaaa tgcactgacc
     4981 tcccacattc cctttttagt aaaatattca gaaataattt aaatacatca ttgcaatgaa
     5041 aataaatgtt ttttattagg cagaatccag atgctcaagg cccttcataa tatcccccag
     5101 tttagtagtt ggacttaggg aacaaaggaa cctttaatag aaattggaca gcaagaaagc
     5161 gagcttctag cttatcctca gtcctgctcc tctgccacaa agtgcacgca gttgccggcc
     5221 gggtcgcgca gggcgaactc ccgcccccac ggctgctcgc cgatctcggt catggccggc
     5281 ccggaggcgt cccggaagtt cgtggacacg acctccgacc actcggcgta cagctcgtcc
     5341 aggccgcgca cccacaccca ggccagggtg ttgtccggca ccacctggtc ctggaccgcg
     5401 ctgatgaaca gggtcacgtc gtcccggacc acaccggcga agtcgtcctc cacgaagtcc
     5461 cgggagaacc cgagccggtc ggtccagaac tcgaccgctc cggcgacgtc gcgcgcggtg
     5521 agcaccggaa cggcactggt caacttggcc atgatggctc ctcctgtcag gagaggaaag
     5581 agaagaaggt tagtacaatt gctatagtga gttgtattat actatgcaga tatactatgc
     5641 caatgattaa ttgtcaaact agggctgcag ggttcatagt gccacttttc ctgcactgcc
     5701 ccatctcctg cccacccttt cccaggcata gacagtcagt gacttaccaa actcacagga
     5761 gggagaaggc agaagcttga gacagacccg cgggaccgcc gaactgcgag gggacgtggc
     5821 tagggcggct tcttttatgg tgcgccggcc ctcggaggca gggcgctcgg ggaggcctag
     5881 cggccaatct gcggtggcag gaggcggggc cgaaggccgt gcctgaccaa tccggagcac
     5941 ataggagtct cagccccccg ccccaaagca aggggaagtc acgcgcctgt agcgccagcg
     6001 tgttgtgaaa tgggggcttg ggggggttgg ggccctgact agtcaaaaca aactcccatt
     6061 gacgtcaatg gggtggagac ttggaaatcc ccgtgagtca aaccgctatc cacgcccatt
     6121 gatgtactgc caaaaccgca tcatcatggt aatagcgatg actaatacgt agatgtactg
     6181 ccaagtagga aagtcccata aggtcatgta ctgggcataa tgccaggcgg gccatttacc
     6241 gtcattgacg tcaatagggg gcgtacttgg catatgatac acttgatgta ctgccaagtg
     6301 ggcagtttac cgtaaatact ccacccattg acgtcaatgg aaagtcccta ttggcgttac
     6361 tatgggaaca tacgtcatta ttgacgtcaa tgggcggggg tcgttgggcg gtcagccagg
     6421 cgggccattt accgtaagtt atgtaacgcc tgcaggttaa ttaagaacat gtgagcaaaa
     6481 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     6541 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     6601 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     6661 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     6721 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     6781 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     6841 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     6901 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     6961 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     7021 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     7081 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     7141 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat ggctagttaa
     7201 ttaacattta aatcagcggc cgcaataaaa tatctttatt ttcattacat ctgtgtgttg
     7261 gttttttgtg tgaatcgtaa ctaacatacg ctctccatca aaacaaaacg aaacaaaaca
     7321 aactagcaaa ataggctgtc cccagtgcaa gtgcaggtgc ca
//
