LOCUS       sequence-442647-        7365 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 7365)
  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 7365)
  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-ADGRB2 CTFdeltaTA-FLAG
FEATURES             Location/Qualifiers
     source          1..7365
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(2709..2735)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     sig_peptide     complement(2736..2795)
                     /codon_start=1
                     /product="human interleukin-2 signal sequence"
                     /label=IL-2 signal sequence
                     /translation="MYRMQLLSCIALSLALVTNS"
     LTR             2845..3113
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     promoter        complement(3126..3337)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     misc_feature    3374..3465
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     polyA_signal    3476..3524
                     /note="synthetic polyadenylation signal"
     rep_origin      3632..4220
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4121..4140
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        4300..4603
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             5196..5570
                     /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    5584..5978
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     polyA_signal    6082..6203
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6119..6138)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6173..6192
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(6918..6941)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
ORIGIN
        1 ggcattgttc tggacctctc gccaggctca ggaacctgtc tggcggatgg ggtaggggga
       61 ggtggctgaa acgtcatgcc gtaagggttc tgcaaggaac cgtaagccgg tcccaagccc
      121 agcccggaat ggtccacgga aagaaaggat ggatacccct ctgtatgagc gacagtcttg
      181 gccgcgcgtc tcggtgtgcc ttccggccta gcccgtggtg cgtcctctcc accgcctcca
      241 ccgcctccgg gctgcaggct cagagtccgt ctaggcagaa ccatgtaatc gccctcgctg
      301 cctggttccg taggacgcag ccaggtcagg tccaattggc ggaggccacc ctcgccgcac
      361 atatacaccg ggttggcttc ctgaggggga gggggctcac caaggccagg gctagcggcc
      421 atgggtacaa ggatgttgcc gggcaagggt gagaaggaca ggcttccctc ggggcccaca
      481 agacaagact ttggctcttc gtcttcatcc agggacagcc tggacagagt cccagtgatg
      541 gtactcgggt tacaagtatt cacctccttg aacagaactg tctggcaggc aaggtcaaca
      601 tctttctcga agtcggacag gatctgcagc tgtccgttct tgcagctgtc gggggaatcc
      661 tcggattcat cggctctgca cacgcccatc tgacatttca cgacgtcctg cacctcacgg
      721 cggaggaagc agtgaactgc ggtgatcaca aagccctggg cggaattgaa tactgcgaac
      781 agagcctgga acagcactga gcgcctgtca gtcatcgcaa gaacggcact catccaagtc
      841 agagccagga gtggcaggac cacgcaagag ctccacaggc tggccatcgc gtttctagca
      901 gaagcagagc taagcagggg gcttggaaca gccccgcagg cactacaagg cagaagcaga
      961 gacgcccagg ggcacctctc actccctgcc ctctgtttct tggacttgtc agagatccca
     1021 tcccgggcca tcagtttatt gaagacgata ataccgatga gcatgttgac caggacgatc
     1081 acagcggccg ggcctacgaa tgcatagagc agccctccct ccaggctcag ccaacagtag
     1141 ctggaggtgc cgtagccctt tgtgcgtgta aaccccacgc tcacagctac caccagagct
     1201 ggcaggcccc agcccagaca cagaaagcgc ttgcgcacca ggcgtgttct catacggccg
     1261 atcacggcca ggtaagactg ccatgcttcg gtgagcaccc agcagaaact gctgaggaaa
     1321 aagaagtgga ggaacgcagc agtcatagta catacgcctt tactcaggac gcgactctgg
     1381 cccacgagaa tgaggatgtt ggaggccagg atactaagac agaagttcag gagaatgata
     1441 gaacgctcgg acttgataaa gcgccagaaa gccgcataaa tggccaggag ggtgagcaac
     1501 gccatacaag acactgcaca gccgatgacg agcgggacgc tggggctgcc ggccagctcc
     1561 agcgtcaggt ccttgggcgg ctgttggaag taaaggttct cgttggtctg cgcatctttc
     1621 agcgcttcat ccacggtctg gcggccgctc gccgcgttaa tcaccgcggt gcgcaccgca
     1681 taccaaaacg cgctcatctg cggaatgttc ggcataattt cgcctttctg cgcgttttcc
     1741 atggtcgccg caatgcgcgg atctttcgcc agttcttctt catagctttt cagcgccacc
     1801 gcgcccagcg gtttatcttt gttcaccgct tccaggcctt catcggtcag cagatagttt
     1861 tccagaaatt ctttcgccag ttctttgttc gggctcgccg cgttaatgcc cgcgctcagc
     1921 acgcccacaa acggtttgct cggctggcct ttaaaggtcg gcagcacggt cacgccatag
     1981 ttcactttgc tggtatcaat gttgctccac gcccacgggc cgttaatggt catcgcggtt
     2041 tcgcctttgt taaacgccgc ttccgcaatg ctataatcgg tatccgcgtt catatgtttg
     2101 tttttaatca gatccaccag aaaggtcagg cccgctttcg cgcccgcgtt atccacgccc
     2161 acatctttaa tatcatattt gccgttttca tatttaaacg catagccgcc atccgccgca
     2221 atcagcggcc aggtaaaata cggttcctgc aggttaaaca tcagcgcgct tttgcctttc
     2281 gctttcagtt ctttatccag cgccggaatt tcttcccagg ttttcggcgg gttcggcagc
     2341 agatctttgt tataaatcag gctcagcgct tccaccgcaa tcggatacgc aatcagtttg
     2401 ccgttatagc gcaccgcatc ccaggtaaac ggatacagtt tatcctgaaa cgctttatcc
     2461 ggggtaattt ccgccagcag gccgctctgc gcatagccgc caaagcgatc atgcgcccaa
     2521 aaaataatat ccgggccatc gccggtcgcc gccacctgcg gaaatttttc ttccagttta
     2581 tccggatgtt ccacggtcac tttaatgccg gtatcttttt caaatttttt gcccacttcc
     2641 gccaggccgt tatagccttt atcgccgtta atccaaatca ccagtttgcc ttcttcaatt
     2701 ttggatccag cgtaatctgg aacatcgtat gggtacgaat tcgtgacaag tgcaagactt
     2761 agtgcaatgc aagacaggag ttgcatcctg tacatggtgg ccctccttcg ccggtgatct
     2821 caggtaggcg ccggtcacag cttggatctg taacggcgca gaacagaaaa cgaaacaaag
     2881 acgtagagtt gagcaagcag ggtcaggcaa agcgtggaga gccggctgag tctaggtagg
     2941 ctccaaggga gcgccggaca aaggcccggt ctcgacctga gctttaaact tacctagacg
     3001 gcggacgcag ttcaggaggc accacaggcg ggaggcggca gaacgcgact caaccggcgt
     3061 ggatggcggc ctcaggtagg gcggcgggcg cgtgaaggag agatgcgagc ccctcgaagc
     3121 ttcagctgtg ttctggcggc aaacccgttg cgaaaaagaa cgttcacggc gactactgca
     3181 cttatatacg gttctccccc accctcggga aaaaggcgga gccagtacac gacatcactt
     3241 tcccagttta ccccgcgcca ccttctctag gcacccgttc aattgccgac ccctcccccc
     3301 aacttctcgg ggactgtggg cgatgtgcgc tctgcccact gacgggcacc ggagcgatcg
     3361 cagatccttc gatagagaaa tgttctggca cctgcacttg cactggggac agcctatttt
     3421 gctagtttgt tttgtttcgt tttgttttga tggagagcgt atgttagtta cgattcacac
     3481 aaaaaaccaa cacacagatg taatgaaaat aaagatattt tattgcggcc gctgatttaa
     3541 atgttaatta actagccatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     3601 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     3661 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     3721 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     3781 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     3841 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     3901 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     3961 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     4021 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     4081 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     4141 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     4201 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     4261 gctggccttt tgctcacatg ttcttaatta acctgcaggc gttacataac ttacggtaaa
     4321 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
     4381 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
     4441 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
     4501 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
     4561 tacttggcag tacatctacg tattagtcat cgctattacc atgatgatgc ggttttggca
     4621 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
     4681 tgacgtcaat gggagtttgt tttgactagt cagggcccca acccccccaa gcccccattt
     4741 cacaacacgc tggcgctaca ggcgcgtgac ttccccttgc tttggggcgg ggggctgaga
     4801 ctcctatgtg ctccggattg gtcaggcacg gccttcggcc ccgcctcctg ccaccgcaga
     4861 ttggccgcta ggcctccccg agcgccctgc ctccgagggc cggcgcacca taaaagaagc
     4921 cgccctagcc acgtcccctc gcagttcggc ggtcccgcgg gtctgtctca agcttctgcc
     4981 ttctccctcc tgtgagtttg gtaagtcact gactgtctat gcctgggaaa gggtgggcag
     5041 gagatggggc agtgcaggaa aagtggcact atgaaccctg cagccctagt ttgacaatta
     5101 atcattggca tagtatatct gcatagtata atacaactca ctatagcaat tgtactaacc
     5161 ttcttctctt tcctctcctg acaggaggag ccatcatggc caagttgacc agtgccgttc
     5221 cggtgctcac cgcgcgcgac gtcgccggag cggtcgagtt ctggaccgac cggctcgggt
     5281 tctcccggga cttcgtggag gacgacttcg ccggtgtggt ccgggacgac gtgaccctgt
     5341 tcatcagcgc ggtccaggac caggtggtgc cggacaacac cctggcctgg gtgtgggtgc
     5401 gcggcctgga cgagctgtac gccgagtggt cggaggtcgt gtccacgaac ttccgggacg
     5461 cctccgggcc ggccatgacc gagatcggcg agcagccgtg ggggcgggag ttcgccctgc
     5521 gcgacccggc cggcaactgc gtgcactttg tggcagagga gcaggactga ggataagcta
     5581 gaagctcgct ttcttgctgt ccaatttcta ttaaaggttc ctttgttccc taagtccaac
     5641 tactaaactg ggggatatta tgaagggcct tgagcatctg gattctgcct aataaaaaac
     5701 atttattttc attgcaatga tgtatttaaa ttatttctga atattttact aaaaagggaa
     5761 tgtgggaggt cagtgcattt aaaacataaa gaaatgaaga gctagttcaa accttgggaa
     5821 aatacactat atcttaaact ccatgaaaga aggtgaggct gcaaacagct aatgcacatt
     5881 ggcaacagcc cctgatgcct atgccttatt catccctcag aaaaggattc aagtagaggc
     5941 ttgatttgga ggttaaagtt ttgctatgct gtattttaga attaattcca taccacattt
     6001 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa
     6061 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
     6121 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     6181 tccaaactca tcaatgtatc ttatcatgtc tggccagcta gcactcattt acccggagac
     6241 agggagaggc tcttctgcgt gtagtggttg tgcagagcct cgtgcatcac ggagcatgag
     6301 aagacgttcc cctgctgcca cctgctcttg tccacggtga gcttgctgta gaggaagaag
     6361 gagccgtcgg agtccagcac gggaggcgtg gtcttgtagt tgttctccgg ctgcccattg
     6421 ctctcccact ccacggcgat gtcgctggga tagaagcctt tgaccaggca ggtcaggctg
     6481 acctggttct tggtcatctc ctcccgggat gggggcaggg tgtacacctg tggttctcgg
     6541 ggctgccctt tggctttgga gatggttttc tcgatggggg ctgggagggc tttgttggag
     6601 accttgcact tgtactcctt gccattcagc cagtcctggt gcaggacggt gaggacgctg
     6661 accacacggt acgtgctgtt gtactgctcc tcccgcggct ttgtcttggc attatgcacc
     6721 tccacgccgt ccacgtacca gttgaacttg acctcagggt cttcgtggct cacgtccacc
     6781 accacgcatg tgacctcagg ggtccgggag atcatgaggg tgtccttggg ttttgggggg
     6841 aagaggaaga ctgacggtcc ccccaggagt tcaggtgctg ggcacggtgg gcatgtgtga
     6901 gttttgtcag atcttcactt gtcgtcatcg tctttgtagt cagaaccaac ctcggtttga
     6961 aaatcaccgt ctggcggttc ggtaggctcc cacgcggcag cacggtgcag cgtcagacgt
     7021 tcgcgtggct taggtggcaa gctacccaaa gtcatggatt taaatgtgct ccagctctga
     7081 tggcgcctgt gctgggacag ggaggggcgc tctccagggc ttggtttatc ggtacatacg
     7141 ctacgctcgg cagcacctcc agagctgaca gaccagcgct tctcccgctt ggcggtggac
     7201 tgggaccggt accggtcgaa tgtgtgaaat ttctggttca gctcatgata cagctcggag
     7261 tgtcttttgc gggtatgcat caccttctcg aaatccaggt cactgtagcg cagtttcttt
     7321 ctttcaagac ttcccatctt cattgtggac ccgggtacgg tccgt
//
