LOCUS       sequence-430005-        6249 bp    DNA     circular SYN 15-APR-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 6249)
  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 6249)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-ADGRE1 CTFdeltaTA-FLAG
FEATURES             Location/Qualifiers
     source          1..6249
                     /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="ENLYFQA"
     CDS             2676..2699
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    3414..3535
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3425..3444)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3479..3498
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     polyA_signal    3639..4033
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     CDS             complement(4047..4421)
                     /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        5014..5317
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(5397..5985)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5477..5496)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    6093..6141
                     /note="synthetic polyadenylation signal"
     misc_feature    6152..6243
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     6192..6211
                     /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 ccaagcttct ggcgaactga
     1801 ccatggactt cagcctttac attatctctc acgttggcat tatcatttcc ttggtgtgcc
     1861 tcgtcctggc tatcgctaca ttcctcctgt gtagatctat ccgcaaccat aacacttacc
     1921 tccacctgca cctctgtgtg tgtctgcttc tggccaagac cctgtttctc gccgggattc
     1981 acaagaccga caacaaaatg ggttgtgcta ttatcgctgg cttcctgcat tatctcttcc
     2041 tggcgtgctt tttctggatg ctggtggagg ctgtcatcct gttcctgatg gtcaggaacc
     2101 tcaaagtagt caactatttc agctcccgga acatcaaaat gttgcacatc tgcgcgttcg
     2161 gctacggcct cccgatgctg gtcgtagtga tttccgcttc cgtccagccc cagggctacg
     2221 gcatgcataa cagatgctgg ctgaacacgg aaacaggttt tatctggtct ttcctgggac
     2281 cagtttgcac tgttatcgtg atcaacagtc tcttgctcac ctggactctg tggatcctcc
     2341 ggcaaaggtt gtccagcgtg aacgctgagg tttccaccct gaaggacacc cgcctcctga
     2401 ccttcaaggc tttcgctcag ctctttatcc tgggctgcag ctgggtcctc ggcatctttc
     2461 agatcggccc cgtggcagga gtcatggcgt acctgtttac cattatcaac tccctgcagg
     2521 gcgcattcat cttccttatt cactgcctcc tgaatggaca ggttcgtgaa gagtacaaaa
     2581 gatggatcac tggcaagacc aagccgagct ctcagagcca aacatctaga atcctgctca
     2641 gcagtatgcc tagtgctagt aagacgggcg gttctgacta caaagacgat gacgacaagt
     2701 gaagatctga caaaactcac acatgcccac cgtgcccagc acctgaactc ctggggggac
     2761 cgtcagtctt cctcttcccc ccaaaaccca aggacaccct catgatctcc cggacccctg
     2821 aggtcacatg cgtggtggtg gacgtgagcc acgaagaccc tgaggtcaag ttcaactggt
     2881 acgtggacgg cgtggaggtg cataatgcca agacaaagcc gcgggaggag cagtacaaca
     2941 gcacgtaccg tgtggtcagc gtcctcaccg tcctgcacca ggactggctg aatggcaagg
     3001 agtacaagtg caaggtctcc aacaaagccc tcccagcccc catcgagaaa accatctcca
     3061 aagccaaagg gcagccccga gaaccacagg tgtacaccct gcccccatcc cgggaggaga
     3121 tgaccaagaa ccaggtcagc ctgacctgcc tggtcaaagg cttctatccc agcgacatcg
     3181 ccgtggagtg ggagagcaat gggcagccgg agaacaacta caagaccacg cctcccgtgc
     3241 tggactccga cggctccttc ttcctctaca gcaagctcac cgtggacaag agcaggtggc
     3301 agcaggggaa cgtcttctca tgctccgtga tgcacgaggc tctgcacaac cactacacgc
     3361 agaagagcct ctccctgtct ccgggtaaat gagtgctagc tggccagaca tgataagata
     3421 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
     3481 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa
     3541 caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag
     3601 caagtaaaac ctctacaaat gtggtatgga attaattcta aaatacagca tagcaaaact
     3661 ttaacctcca aatcaagcct ctacttgaat ccttttctga gggatgaata aggcataggc
     3721 atcaggggct gttgccaatg tgcattagct gtttgcagcc tcaccttctt tcatggagtt
     3781 taagatatag tgtattttcc caaggtttga actagctctt catttcttta tgttttaaat
     3841 gcactgacct cccacattcc ctttttagta aaatattcag aaataattta aatacatcat
     3901 tgcaatgaaa ataaatgttt tttattaggc agaatccaga tgctcaaggc ccttcataat
     3961 atcccccagt ttagtagttg gacttaggga acaaaggaac ctttaataga aattggacag
     4021 caagaaagcg agcttctagc ttatcctcag tcctgctcct ctgccacaaa gtgcacgcag
     4081 ttgccggccg ggtcgcgcag ggcgaactcc cgcccccacg gctgctcgcc gatctcggtc
     4141 atggccggcc cggaggcgtc ccggaagttc gtggacacga cctccgacca ctcggcgtac
     4201 agctcgtcca ggccgcgcac ccacacccag gccagggtgt tgtccggcac cacctggtcc
     4261 tggaccgcgc tgatgaacag ggtcacgtcg tcccggacca caccggcgaa gtcgtcctcc
     4321 acgaagtccc gggagaaccc gagccggtcg gtccagaact cgaccgctcc ggcgacgtcg
     4381 cgcgcggtga gcaccggaac ggcactggtc aacttggcca tgatggctcc tcctgtcagg
     4441 agaggaaaga gaagaaggtt agtacaattg ctatagtgag ttgtattata ctatgcagat
     4501 atactatgcc aatgattaat tgtcaaacta gggctgcagg gttcatagtg ccacttttcc
     4561 tgcactgccc catctcctgc ccaccctttc ccaggcatag acagtcagtg acttaccaaa
     4621 ctcacaggag ggagaaggca gaagcttgag acagacccgc gggaccgccg aactgcgagg
     4681 ggacgtggct agggcggctt cttttatggt gcgccggccc tcggaggcag ggcgctcggg
     4741 gaggcctagc ggccaatctg cggtggcagg aggcggggcc gaaggccgtg cctgaccaat
     4801 ccggagcaca taggagtctc agccccccgc cccaaagcaa ggggaagtca cgcgcctgta
     4861 gcgccagcgt gttgtgaaat gggggcttgg gggggttggg gccctgacta gtcaaaacaa
     4921 actcccattg acgtcaatgg ggtggagact tggaaatccc cgtgagtcaa accgctatcc
     4981 acgcccattg atgtactgcc aaaaccgcat catcatggta atagcgatga ctaatacgta
     5041 gatgtactgc caagtaggaa agtcccataa ggtcatgtac tgggcataat gccaggcggg
     5101 ccatttaccg tcattgacgt caataggggg cgtacttggc atatgataca cttgatgtac
     5161 tgccaagtgg gcagtttacc gtaaatactc cacccattga cgtcaatgga aagtccctat
     5221 tggcgttact atgggaacat acgtcattat tgacgtcaat gggcgggggt cgttgggcgg
     5281 tcagccaggc gggccattta ccgtaagtta tgtaacgcct gcaggttaat taagaacatg
     5341 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     5401 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     5461 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     5521 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     5581 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     5641 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     5701 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     5761 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     5821 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     5881 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     5941 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     6001 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     6061 gctagttaat taacatttaa atcagcggcc gcaataaaat atctttattt tcattacatc
     6121 tgtgtgttgg ttttttgtgt gaatcgtaac taacatacgc tctccatcaa aacaaaacga
     6181 aacaaaacaa actagcaaaa taggctgtcc ccagtgcaag tgcaggtgcc agaacatttc
     6241 tctatcgaa
//
