LOCUS       sequence-430301-        6258 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 6258)
  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 6258)
  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-ADGRE5 CTFdeltaTA-FLAG
FEATURES             Location/Qualifiers
     source          1..6258
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(559..585)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     sig_peptide     complement(586..645)
                     /codon_start=1
                     /product="human interleukin-2 signal sequence"
                     /label=IL-2 signal sequence
                     /translation="MYRMQLLSCIALSLALVTNS"
     LTR             695..963
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     promoter        complement(976..1187)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     misc_feature    1224..1315
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     complement(1256..1275)
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     polyA_signal    1326..1374
                     /note="synthetic polyadenylation signal"
     rep_origin      1482..2070
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1971..1990
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2150..2453
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             3046..3420
                     /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"
     polyA_signal    3434..3828
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     polyA_signal    3932..4053
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3969..3988)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4023..4042
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(4768..4791)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(5689..5709)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQA"
ORIGIN
        1 atccacgccc acatctttaa tatcatattt gccgttttca tatttaaacg catagccgcc
       61 atccgccgca atcagcggcc aggtaaaata cggttcctgc aggttaaaca tcagcgcgct
      121 tttgcctttc gctttcagtt ctttatccag cgccggaatt tcttcccagg ttttcggcgg
      181 gttcggcagc agatctttgt tataaatcag gctcagcgct tccaccgcaa tcggatacgc
      241 aatcagtttg ccgttatagc gcaccgcatc ccaggtaaac ggatacagtt tatcctgaaa
      301 cgctttatcc ggggtaattt ccgccagcag gccgctctgc gcatagccgc caaagcgatc
      361 atgcgcccaa aaaataatat ccgggccatc gccggtcgcc gccacctgcg gaaatttttc
      421 ttccagttta tccggatgtt ccacggtcac tttaatgccg gtatcttttt caaatttttt
      481 gcccacttcc gccaggccgt tatagccttt atcgccgtta atccaaatca ccagtttgcc
      541 ttcttcaatt ttggatccag cgtaatctgg aacatcgtat gggtacgaat tcgtgacaag
      601 tgcaagactt agtgcaatgc aagacaggag ttgcatcctg tacatggtgg ccctccttcg
      661 ccggtgatct caggtaggcg ccggtcacag cttggatctg taacggcgca gaacagaaaa
      721 cgaaacaaag acgtagagtt gagcaagcag ggtcaggcaa agcgtggaga gccggctgag
      781 tctaggtagg ctccaaggga gcgccggaca aaggcccggt ctcgacctga gctttaaact
      841 tacctagacg gcggacgcag ttcaggaggc accacaggcg ggaggcggca gaacgcgact
      901 caaccggcgt ggatggcggc ctcaggtagg gcggcgggcg cgtgaaggag agatgcgagc
      961 ccctcgaagc ttcagctgtg ttctggcggc aaacccgttg cgaaaaagaa cgttcacggc
     1021 gactactgca cttatatacg gttctccccc accctcggga aaaaggcgga gccagtacac
     1081 gacatcactt tcccagttta ccccgcgcca ccttctctag gcacccgttc aattgccgac
     1141 ccctcccccc aacttctcgg ggactgtggg cgatgtgcgc tctgcccact gacgggcacc
     1201 ggagcgatcg cagatccttc gatagagaaa tgttctggca cctgcacttg cactggggac
     1261 agcctatttt gctagtttgt tttgtttcgt tttgttttga tggagagcgt atgttagtta
     1321 cgattcacac aaaaaaccaa cacacagatg taatgaaaat aaagatattt tattgcggcc
     1381 gctgatttaa atgttaatta actagccatg accaaaatcc cttaacgtga gttttcgttc
     1441 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     1501 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     1561 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     1621 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     1681 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     1741 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     1801 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     1861 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     1921 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     1981 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     2041 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     2101 ctggcctttt gctggccttt tgctcacatg ttcttaatta acctgcaggc gttacataac
     2161 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
     2221 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
     2281 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
     2341 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat
     2401 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atgatgatgc
     2461 ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc
     2521 tccaccccat tgacgtcaat gggagtttgt tttgactagt cagggcccca acccccccaa
     2581 gcccccattt cacaacacgc tggcgctaca ggcgcgtgac ttccccttgc tttggggcgg
     2641 ggggctgaga ctcctatgtg ctccggattg gtcaggcacg gccttcggcc ccgcctcctg
     2701 ccaccgcaga ttggccgcta ggcctccccg agcgccctgc ctccgagggc cggcgcacca
     2761 taaaagaagc cgccctagcc acgtcccctc gcagttcggc ggtcccgcgg gtctgtctca
     2821 agcttctgcc ttctccctcc tgtgagtttg gtaagtcact gactgtctat gcctgggaaa
     2881 gggtgggcag gagatggggc agtgcaggaa aagtggcact atgaaccctg cagccctagt
     2941 ttgacaatta atcattggca tagtatatct gcatagtata atacaactca ctatagcaat
     3001 tgtactaacc ttcttctctt tcctctcctg acaggaggag ccatcatggc caagttgacc
     3061 agtgccgttc cggtgctcac cgcgcgcgac gtcgccggag cggtcgagtt ctggaccgac
     3121 cggctcgggt tctcccggga cttcgtggag gacgacttcg ccggtgtggt ccgggacgac
     3181 gtgaccctgt tcatcagcgc ggtccaggac caggtggtgc cggacaacac cctggcctgg
     3241 gtgtgggtgc gcggcctgga cgagctgtac gccgagtggt cggaggtcgt gtccacgaac
     3301 ttccgggacg cctccgggcc ggccatgacc gagatcggcg agcagccgtg ggggcgggag
     3361 ttcgccctgc gcgacccggc cggcaactgc gtgcactttg tggcagagga gcaggactga
     3421 ggataagcta gaagctcgct ttcttgctgt ccaatttcta ttaaaggttc ctttgttccc
     3481 taagtccaac tactaaactg ggggatatta tgaagggcct tgagcatctg gattctgcct
     3541 aataaaaaac atttattttc attgcaatga tgtatttaaa ttatttctga atattttact
     3601 aaaaagggaa tgtgggaggt cagtgcattt aaaacataaa gaaatgaaga gctagttcaa
     3661 accttgggaa aatacactat atcttaaact ccatgaaaga aggtgaggct gcaaacagct
     3721 aatgcacatt ggcaacagcc cctgatgcct atgccttatt catccctcag aaaaggattc
     3781 aagtagaggc ttgatttgga ggttaaagtt ttgctatgct gtattttaga attaattcca
     3841 taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct
     3901 gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta
     3961 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     4021 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggccagcta gcactcattt
     4081 acccggagac agggagaggc tcttctgcgt gtagtggttg tgcagagcct cgtgcatcac
     4141 ggagcatgag aagacgttcc cctgctgcca cctgctcttg tccacggtga gcttgctgta
     4201 gaggaagaag gagccgtcgg agtccagcac gggaggcgtg gtcttgtagt tgttctccgg
     4261 ctgcccattg ctctcccact ccacggcgat gtcgctggga tagaagcctt tgaccaggca
     4321 ggtcaggctg acctggttct tggtcatctc ctcccgggat gggggcaggg tgtacacctg
     4381 tggttctcgg ggctgccctt tggctttgga gatggttttc tcgatggggg ctgggagggc
     4441 tttgttggag accttgcact tgtactcctt gccattcagc cagtcctggt gcaggacggt
     4501 gaggacgctg accacacggt acgtgctgtt gtactgctcc tcccgcggct ttgtcttggc
     4561 attatgcacc tccacgccgt ccacgtacca gttgaacttg acctcagggt cttcgtggct
     4621 cacgtccacc accacgcatg tgacctcagg ggtccgggag atcatgaggg tgtccttggg
     4681 ttttgggggg aagaggaaga ctgacggtcc ccccaggagt tcaggtgctg ggcacggtgg
     4741 gcatgtgtga gttttgtcag atcttcactt gtcgtcatcg tctttgtagt cagaacctat
     4801 gccggactct gatgccctga gggcccgggt ctgattgtgg ccagtgccag acgtggtgga
     4861 ggtgaattct gagtacttgc tccccccagc aactaggcag gcccacttcc ggtattcttc
     4921 ccgaaccttc ttgttgagca ggcagtgcag caggtagagg aaggcgccct gcaggcagtt
     4981 gaggatggta aacacatagg tcagcaccaa gctccgatcg tcgaagatga acaggccaaa
     5041 gacccaggtg cagcccaaca ggaagagctg cgcgatggcc gtgatggtca gcgccctcgc
     5101 cttctttaat ttcttcatgt ctggattgat ttcagaaaac ttctgagtga gcttccagac
     5161 ggtagtcacg aaaatgacag cattgcacaa aatgatgaag gtcacaggtc ccaagaagct
     5221 ccagaggaag ccctgctcaa agtccaacca gcagtatctg gggcggccgt agcccttgct
     5281 gtagatggca gccgagacgc ccacgatgag caggggcacg ccatagccga tcaggcagag
     5341 ccagcgcgta ctcaggccct ggccttggaa cacgcgcacc acaagaaagt agagctccag
     5401 gccttcgagg ctcatccagc agaaggcggc caggaaacag tagtgcagca gcccggccac
     5461 caggcggcag cgcagcccca cctggccgcc ttcgttctcg atgccggcca ggaagatggt
     5521 ggagcccacg aagaggcaga tgcagaggtg caggtgtatg gtggtgcgcg agccctggat
     5581 gggccgcacc agcaggaaag tgaggatgca cagcagcagg cagaagagtg acagcgccag
     5641 tcccaccctg gtgatcaggg tcagcttcca gtcctccacg tcataatgag cttggaagta
     5701 aaggttctcg ttggtctgcg catctttcag cgcttcatcc acggtctggc ggccgctcgc
     5761 cgcgttaatc accgcggtgc gcaccgcata ccaaaacgcg ctcatctgcg gaatgttcgg
     5821 cataatttcg cctttctgcg cgttttccat ggtcgccgca atgcgcggat ctttcgccag
     5881 ttcttcttca tagcttttca gcgccaccgc gcccagcggt ttatctttgt tcaccgcttc
     5941 caggccttca tcggtcagca gatagttttc cagaaattct ttcgccagtt ctttgttcgg
     6001 gctcgccgcg ttaatgcccg cgctcagcac gcccacaaac ggtttgctcg gctggccttt
     6061 aaaggtcggc agcacggtca cgccatagtt cactttgctg gtatcaatgt tgctccacgc
     6121 ccacgggccg ttaatggtca tcgcggtttc gcctttgtta aacgccgctt ccgcaatgct
     6181 ataatcggta tccgcgttca tatgtttgtt tttaatcaga tccaccagaa aggtcaggcc
     6241 cgctttcgcg cccgcgtt
//
