LOCUS       sequence-430274-        7065 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 7065)
  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 7065)
  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-ADGRC2 CTF-FLAG
FEATURES             Location/Qualifiers
     source          1..7065
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    342..736
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     polyA_signal    840..961
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(877..896)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     931..950
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(1676..1699)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(4532..4558)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     sig_peptide     complement(4559..4618)
                     /codon_start=1
                     /product="human interleukin-2 signal sequence"
                     /label=IL-2 signal sequence
                     /translation="MYRMQLLSCIALSLALVTNS"
     LTR             4668..4936
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     promoter        complement(4949..5160)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     misc_feature    5197..5288
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     complement(5229..5248)
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     polyA_signal    5299..5347
                     /note="synthetic polyadenylation signal"
     rep_origin      5455..6043
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5944..5963
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        6123..6426
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             join(7019..7065,1..328)
                     /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"
ORIGIN
        1 cgccggagcg gtcgagttct ggaccgaccg gctcgggttc tcccgggact tcgtggagga
       61 cgacttcgcc ggtgtggtcc gggacgacgt gaccctgttc atcagcgcgg tccaggacca
      121 ggtggtgccg gacaacaccc tggcctgggt gtgggtgcgc ggcctggacg agctgtacgc
      181 cgagtggtcg gaggtcgtgt ccacgaactt ccgggacgcc tccgggccgg ccatgaccga
      241 gatcggcgag cagccgtggg ggcgggagtt cgccctgcgc gacccggccg gcaactgcgt
      301 gcactttgtg gcagaggagc aggactgagg ataagctaga agctcgcttt cttgctgtcc
      361 aatttctatt aaaggttcct ttgttcccta agtccaacta ctaaactggg ggatattatg
      421 aagggccttg agcatctgga ttctgcctaa taaaaaacat ttattttcat tgcaatgatg
      481 tatttaaatt atttctgaat attttactaa aaagggaatg tgggaggtca gtgcatttaa
      541 aacataaaga aatgaagagc tagttcaaac cttgggaaaa tacactatat cttaaactcc
      601 atgaaagaag gtgaggctgc aaacagctaa tgcacattgg caacagcccc tgatgcctat
      661 gccttattca tccctcagaa aaggattcaa gtagaggctt gatttggagg ttaaagtttt
      721 gctatgctgt attttagaat taattccata ccacatttgt agaggtttta cttgctttaa
      781 aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta
      841 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
      901 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
      961 atcatgtctg gccagctagc actcatttac ccggagacag ggagaggctc ttctgcgtgt
     1021 agtggttgtg cagagcctcg tgcatcacgg agcatgagaa gacgttcccc tgctgccacc
     1081 tgctcttgtc cacggtgagc ttgctgtaga ggaagaagga gccgtcggag tccagcacgg
     1141 gaggcgtggt cttgtagttg ttctccggct gcccattgct ctcccactcc acggcgatgt
     1201 cgctgggata gaagcctttg accaggcagg tcaggctgac ctggttcttg gtcatctcct
     1261 cccgggatgg gggcagggtg tacacctgtg gttctcgggg ctgccctttg gctttggaga
     1321 tggttttctc gatgggggct gggagggctt tgttggagac cttgcacttg tactccttgc
     1381 cattcagcca gtcctggtgc aggacggtga ggacgctgac cacacggtac gtgctgttgt
     1441 actgctcctc ccgcggcttt gtcttggcat tatgcacctc cacgccgtcc acgtaccagt
     1501 tgaacttgac ctcagggtct tcgtggctca cgtccaccac cacgcatgtg acctcagggg
     1561 tccgggagat catgagggtg tccttgggtt ttggggggaa gaggaagact gacggtcccc
     1621 ccaggagttc aggtgctggg cacggtgggc atgtgtgagt tttgtcagat cttcacttgt
     1681 cgtcatcgtc tttgtagtca gaaccatgca ggaagttgaa aaaaagaaat tcggaaccgg
     1741 aactatcttc gtcgacggtc cccgccttaa tagacatggc aataggcatc acgccgttca
     1801 gttgctcctg caaagactgg cgcggagggg gccgaggcgg aggacctccc cgggagcctt
     1861 cggatgcaga actaccgcga gagctgccag tgcactgttc cagaggcaat ctcagcaaag
     1921 aggacttctc ggagatagtt gggagacatt tctttttcaa gatgccctta tggggttgag
     1981 cactgctacc tggcaaaggg cccaggctac cttcacggct cagggcatcg ccgttctccc
     2041 taagacgttc ctctggggcg ccgttgccgg agctctcctt ggcggtcgtg ccgaaatccc
     2101 caggccaggg ggccttaccc ggtccgggac cgccgtcttt cggagtgcta tggagaggca
     2161 ggcgttcagc gccggggccc agaagggaat cccagccctg ctctcctggg aaggccgcct
     2221 cttcctcttc ttcctcttcc tcctcttcgc tgtcagagga atgggtagag gcgtatgacc
     2281 cagactggtc atcctcgagg gacaaatcgg aatcggagtc ggtatcaggg tcatgctgtt
     2341 ggtcctgtcc ctccaggaag agtgatccgg ggtcacccag gcccggtgga ccttggcccg
     2401 ggttcagggc ggactcttcg cgcaggagga acgggatgta ggatggctgg gacttgcctg
     2461 accggctagt ggagtgcagg gacccggcag aatcaccata tggctgatag agtctgccat
     2521 cagcgtaggg gctagggcag ttgtagctac tggtgagagt ggacttggta gtcagggcgg
     2581 gatctgggct aggcttgcgg ctgcaggcga gcttcagagc ctttctgact tctttggaca
     2641 gaaccacgta agacaaaaag atgaaggggc cctggataca gttacaggta gcgaaaagat
     2701 agtgaaacag caacgtgtcg ctattgacgc tcaggagtgc gagaagccat gtggcggaga
     2761 gcagcaacag aacggcgaag gaaggctgca gtccgcttac tgggcctttc ttttcgaatc
     2821 cctgcctctg tgcggcacag gaggcccggg ctgcgaggat gtaaaggaag acggacatgg
     2881 acacagcgaa cgccactggt ccggcgaagg accagatcag ggtgtcgtag atggagagcc
     2941 aacagaagtc ggggttcccg taaccctcgg ggtccagacc aacggcgagg ccggtgatga
     3001 acgctgggac gccccagccc aacatgtaat agaagcgcat aggtcctgtg ttgacatcgc
     3061 gaacctctgt aagagccctg tacaggtgca gagcctccag gagggcccaa gagaaggtgc
     3121 acaaatacag gaagtgcagc aagatcgcga tcactgtgca ggcgaagggc aggtctgcct
     3181 ggttgatgcc caggaggaag acgagctggg ccaatcccag ggccgctgtc aggttcctac
     3241 gaataccatg ctggttagaa cggaggatcc tcagcaaggt caggaaaaag aaggtcagga
     3301 gcaatgcggc gagagtgact ccgagcgcca cgtaggtcaa agttttcaga ggcaggatct
     3361 cgccgttttc gcgacgggaa acgtccatga gtacggcgaa tgatgtttgg aagtaaaggt
     3421 tctcgttggt ctgcgcatct ttcagcgctt catccacggt ctggcggccg ctcgccgcgt
     3481 taatcaccgc ggtgcgcacc gcataccaaa acgcgctcat ctgcggaatg ttcggcataa
     3541 tttcgccttt ctgcgcgttt tccatggtcg ccgcaatgcg cggatctttc gccagttctt
     3601 cttcatagct tttcagcgcc accgcgccca gcggtttatc tttgttcacc gcttccaggc
     3661 cttcatcggt cagcagatag ttttccagaa attctttcgc cagttctttg ttcgggctcg
     3721 ccgcgttaat gcccgcgctc agcacgccca caaacggttt gctcggctgg cctttaaagg
     3781 tcggcagcac ggtcacgcca tagttcactt tgctggtatc aatgttgctc cacgcccacg
     3841 ggccgttaat ggtcatcgcg gtttcgcctt tgttaaacgc cgcttccgca atgctataat
     3901 cggtatccgc gttcatatgt ttgtttttaa tcagatccac cagaaaggtc aggcccgctt
     3961 tcgcgcccgc gttatccacg cccacatctt taatatcata tttgccgttt tcatatttaa
     4021 acgcatagcc gccatccgcc gcaatcagcg gccaggtaaa atacggttcc tgcaggttaa
     4081 acatcagcgc gcttttgcct ttcgctttca gttctttatc cagcgccgga atttcttccc
     4141 aggttttcgg cgggttcggc agcagatctt tgttataaat caggctcagc gcttccaccg
     4201 caatcggata cgcaatcagt ttgccgttat agcgcaccgc atcccaggta aacggataca
     4261 gtttatcctg aaacgcttta tccggggtaa tttccgccag caggccgctc tgcgcatagc
     4321 cgccaaagcg atcatgcgcc caaaaaataa tatccgggcc atcgccggtc gccgccacct
     4381 gcggaaattt ttcttccagt ttatccggat gttccacggt cactttaatg ccggtatctt
     4441 tttcaaattt tttgcccact tccgccaggc cgttatagcc tttatcgccg ttaatccaaa
     4501 tcaccagttt gccttcttca attttggatc cagcgtaatc tggaacatcg tatgggtacg
     4561 aattcgtgac aagtgcaaga cttagtgcaa tgcaagacag gagttgcatc ctgtacatgg
     4621 tggccctcct tcgccggtga tctcaggtag gcgccggtca cagcttggat ctgtaacggc
     4681 gcagaacaga aaacgaaaca aagacgtaga gttgagcaag cagggtcagg caaagcgtgg
     4741 agagccggct gagtctaggt aggctccaag ggagcgccgg acaaaggccc ggtctcgacc
     4801 tgagctttaa acttacctag acggcggacg cagttcagga ggcaccacag gcgggaggcg
     4861 gcagaacgcg actcaaccgg cgtggatggc ggcctcaggt agggcggcgg gcgcgtgaag
     4921 gagagatgcg agcccctcga agcttcagct gtgttctggc ggcaaacccg ttgcgaaaaa
     4981 gaacgttcac ggcgactact gcacttatat acggttctcc cccaccctcg ggaaaaaggc
     5041 ggagccagta cacgacatca ctttcccagt ttaccccgcg ccaccttctc taggcacccg
     5101 ttcaattgcc gacccctccc cccaacttct cggggactgt gggcgatgtg cgctctgccc
     5161 actgacgggc accggagcga tcgcagatcc ttcgatagag aaatgttctg gcacctgcac
     5221 ttgcactggg gacagcctat tttgctagtt tgttttgttt cgttttgttt tgatggagag
     5281 cgtatgttag ttacgattca cacaaaaaac caacacacag atgtaatgaa aataaagata
     5341 ttttattgcg gccgctgatt taaatgttaa ttaactagcc atgaccaaaa tcccttaacg
     5401 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     5461 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     5521 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     5581 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     5641 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     5701 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     5761 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     5821 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     5881 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     5941 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     6001 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     6061 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttcttaa ttaacctgca
     6121 ggcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
     6181 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
     6241 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
     6301 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
     6361 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
     6421 accatgatga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg
     6481 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttgact agtcagggcc
     6541 ccaacccccc caagccccca tttcacaaca cgctggcgct acaggcgcgt gacttcccct
     6601 tgctttgggg cggggggctg agactcctat gtgctccgga ttggtcaggc acggccttcg
     6661 gccccgcctc ctgccaccgc agattggccg ctaggcctcc ccgagcgccc tgcctccgag
     6721 ggccggcgca ccataaaaga agccgcccta gccacgtccc ctcgcagttc ggcggtcccg
     6781 cgggtctgtc tcaagcttct gccttctccc tcctgtgagt ttggtaagtc actgactgtc
     6841 tatgcctggg aaagggtggg caggagatgg ggcagtgcag gaaaagtggc actatgaacc
     6901 ctgcagccct agtttgacaa ttaatcattg gcatagtata tctgcatagt ataatacaac
     6961 tcactatagc aattgtacta accttcttct ctttcctctc ctgacaggag gagccatcat
     7021 ggccaagttg accagtgccg ttccggtgct caccgcgcgc gacgt
//
