LOCUS       sequence-430115-        6411 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 6411)
  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 6411)
  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-ADGRF5 CTFdeltaTA-FLAG
FEATURES             Location/Qualifiers
     source          1..6411
                     /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             2856..2879
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    3594..3715
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3605..3624)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3659..3678
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     polyA_signal    3819..4213
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     CDS             complement(4227..4601)
                     /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        5194..5497
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(5577..6165)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5657..5676)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    6273..6321
                     /note="synthetic polyadenylation signal"
     misc_feature    join(6332..6411,1..12)
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     6372..6391
                     /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 aatcccctga ctccccagat cctagttctc tcctgggaat actcctggat attatttctt
     1861 atgttggggt gggcttttcc atcttgagct tggcagcctg tctagttgtg gaagctgtgg
     1921 tgtggaaatc ggtgaccaag aatcggactt cttatatgcg ccacacctgc atagtgaata
     1981 tcgctgcctc ccttctggtc gccaacacct ggttcattgt ggtcgctgcc atccaggaca
     2041 atcgctacat actctgcaag acagcctgtg tggctgccac cttcttcatc cacttcttct
     2101 acctcagcgt cttcttctgg atgctgacac tgggcctcat gctgttctat cgcctggttt
     2161 tcattctgca tgaaacaagc aggtccactc agaaagccat tgccttctgt cttggctatg
     2221 gctgcccact tgccatctcg gtcatcacgc tgggagccac ccagccccgg gaagtctata
     2281 cgaggaagaa tgtctgttgg ctcaactggg aggacaccaa ggccctgctg gctttcgcca
     2341 tcccagcact gatcattgtg gtggtgaaca taaccatcac tattgtggtc atcaccaaga
     2401 tcctgaggcc ttccattgga gacaagccat gcaagcagga gaagagcagc ctgtttcaga
     2461 tcagcaagag cattggggtc ctcacaccac tcttgggcct cacttggggt tttggtctca
     2521 ccactgtgtt cccagggacc aaccttgtgt tccatatcat atttgccatc ctcaatgtct
     2581 tccagggatt attcatttta ctctttggat gcctctggga tctgaaggta caggaagctt
     2641 tgctgaataa gttttcattg tcgagatggt cttcacagca ctcaaagtca acatccctgg
     2701 gttcatccac acctgtgttt tctatgagtt ctccaatatc aaggagattt aacaatttgt
     2761 ttggtaaaac aggaacgtat aatgtttcca ccccagaagc aaccagctca tccctggaaa
     2821 actcatccag tgcttcttcg ttgctcaacg gttctgacta caaagacgat gacgacaagt
     2881 gaagatctga caaaactcac acatgcccac cgtgcccagc acctgaactc ctggggggac
     2941 cgtcagtctt cctcttcccc ccaaaaccca aggacaccct catgatctcc cggacccctg
     3001 aggtcacatg cgtggtggtg gacgtgagcc acgaagaccc tgaggtcaag ttcaactggt
     3061 acgtggacgg cgtggaggtg cataatgcca agacaaagcc gcgggaggag cagtacaaca
     3121 gcacgtaccg tgtggtcagc gtcctcaccg tcctgcacca ggactggctg aatggcaagg
     3181 agtacaagtg caaggtctcc aacaaagccc tcccagcccc catcgagaaa accatctcca
     3241 aagccaaagg gcagccccga gaaccacagg tgtacaccct gcccccatcc cgggaggaga
     3301 tgaccaagaa ccaggtcagc ctgacctgcc tggtcaaagg cttctatccc agcgacatcg
     3361 ccgtggagtg ggagagcaat gggcagccgg agaacaacta caagaccacg cctcccgtgc
     3421 tggactccga cggctccttc ttcctctaca gcaagctcac cgtggacaag agcaggtggc
     3481 agcaggggaa cgtcttctca tgctccgtga tgcacgaggc tctgcacaac cactacacgc
     3541 agaagagcct ctccctgtct ccgggtaaat gagtgctagc tggccagaca tgataagata
     3601 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
     3661 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa
     3721 caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag
     3781 caagtaaaac ctctacaaat gtggtatgga attaattcta aaatacagca tagcaaaact
     3841 ttaacctcca aatcaagcct ctacttgaat ccttttctga gggatgaata aggcataggc
     3901 atcaggggct gttgccaatg tgcattagct gtttgcagcc tcaccttctt tcatggagtt
     3961 taagatatag tgtattttcc caaggtttga actagctctt catttcttta tgttttaaat
     4021 gcactgacct cccacattcc ctttttagta aaatattcag aaataattta aatacatcat
     4081 tgcaatgaaa ataaatgttt tttattaggc agaatccaga tgctcaaggc ccttcataat
     4141 atcccccagt ttagtagttg gacttaggga acaaaggaac ctttaataga aattggacag
     4201 caagaaagcg agcttctagc ttatcctcag tcctgctcct ctgccacaaa gtgcacgcag
     4261 ttgccggccg ggtcgcgcag ggcgaactcc cgcccccacg gctgctcgcc gatctcggtc
     4321 atggccggcc cggaggcgtc ccggaagttc gtggacacga cctccgacca ctcggcgtac
     4381 agctcgtcca ggccgcgcac ccacacccag gccagggtgt tgtccggcac cacctggtcc
     4441 tggaccgcgc tgatgaacag ggtcacgtcg tcccggacca caccggcgaa gtcgtcctcc
     4501 acgaagtccc gggagaaccc gagccggtcg gtccagaact cgaccgctcc ggcgacgtcg
     4561 cgcgcggtga gcaccggaac ggcactggtc aacttggcca tgatggctcc tcctgtcagg
     4621 agaggaaaga gaagaaggtt agtacaattg ctatagtgag ttgtattata ctatgcagat
     4681 atactatgcc aatgattaat tgtcaaacta gggctgcagg gttcatagtg ccacttttcc
     4741 tgcactgccc catctcctgc ccaccctttc ccaggcatag acagtcagtg acttaccaaa
     4801 ctcacaggag ggagaaggca gaagcttgag acagacccgc gggaccgccg aactgcgagg
     4861 ggacgtggct agggcggctt cttttatggt gcgccggccc tcggaggcag ggcgctcggg
     4921 gaggcctagc ggccaatctg cggtggcagg aggcggggcc gaaggccgtg cctgaccaat
     4981 ccggagcaca taggagtctc agccccccgc cccaaagcaa ggggaagtca cgcgcctgta
     5041 gcgccagcgt gttgtgaaat gggggcttgg gggggttggg gccctgacta gtcaaaacaa
     5101 actcccattg acgtcaatgg ggtggagact tggaaatccc cgtgagtcaa accgctatcc
     5161 acgcccattg atgtactgcc aaaaccgcat catcatggta atagcgatga ctaatacgta
     5221 gatgtactgc caagtaggaa agtcccataa ggtcatgtac tgggcataat gccaggcggg
     5281 ccatttaccg tcattgacgt caataggggg cgtacttggc atatgataca cttgatgtac
     5341 tgccaagtgg gcagtttacc gtaaatactc cacccattga cgtcaatgga aagtccctat
     5401 tggcgttact atgggaacat acgtcattat tgacgtcaat gggcgggggt cgttgggcgg
     5461 tcagccaggc gggccattta ccgtaagtta tgtaacgcct gcaggttaat taagaacatg
     5521 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     5581 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     5641 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     5701 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     5761 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     5821 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     5881 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     5941 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     6001 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     6061 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     6121 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     6181 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     6241 gctagttaat taacatttaa atcagcggcc gcaataaaat atctttattt tcattacatc
     6301 tgtgtgttgg ttttttgtgt gaatcgtaac taacatacgc tctccatcaa aacaaaacga
     6361 aacaaaacaa actagcaaaa taggctgtcc ccagtgcaag tgcaggtgcc a
//
