LOCUS       sequence-430117-        6993 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 6993)
  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 6993)
  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-ADGRA1 CTFdeltaTA-FLAG
FEATURES             Location/Qualifiers
     source          1..6993
                     /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             3438..3461
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    4176..4297
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4187..4206)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4241..4260
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     polyA_signal    4401..4795
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     CDS             complement(4809..5183)
                     /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        5776..6079
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(6159..6747)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6239..6258)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    6855..6903
                     /note="synthetic polyadenylation signal"
     misc_feature    join(6914..6993,1..12)
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     6954..6973
                     /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 aacatcccgt cgtgtacgcc tgcaccgccg ttatgcttct gtgcctcctg gcttctttcg
     1861 tgacctacat tgtccatcag tctgcgatca gaatttcccg caaagggcgc cataccctcc
     1921 tgaacttctg cttccacgca gctctgacct ttacggtgtt cgccggaggc atcaaccgta
     1981 ctaagtaccc aatcctctgt caagcagttg ggattgtcct gcactacagc acccttagta
     2041 ccatgctttg gattggcgta accgcccgta acatttacaa gcaggtcacc aaaaaggcgc
     2101 cactgtgcct ggacacagat caacctccct acccaagaca gcctctgctt agattctacc
     2161 tcgtgagcgg aggcgtgccg tttatcattt gtggagtcac tgccgcaacc aacatccgca
     2221 actacggtac agaggatgag gatactgctt actgttggat ggcatgggaa ccatctctgg
     2281 gcgccttcta cggtcccgct gccattatca cactggtaac gtgtgtgtac ttcctgggca
     2341 cttacgtgca actgcgcaga catccgggca ggcgctacga gctgagaact cagcccgaag
     2401 agcagcgccg gttggctact ccagagggtg gcagaggtat ccgcccaggc acacctccag
     2461 cccacgacgc tccaggtgca agtgtgcttc agaacgaaca ctcattccag gcccaactga
     2521 gggcagcggc ctttactctg ttcctcttca ccgctacctg ggcgtttgga gccctggctg
     2581 tgtcccaagg gcacttcctg gacatggtgt tctcctgtct gtacggcgcg ttctgcgtta
     2641 ccctcggact cttcgtgctg attcatcact gcgccaaacg cgaagacgtt tggcagtgct
     2701 ggtgggcatg ctgtccacct cggaaagacg cgcatcctgc cctggacgct aacggagccg
     2761 ctcttggtcg ggcagcttgc ctgcacagcc ctggactcgg gcagcctcgc ggcttcgctc
     2821 atccacccgg tccgtgtaag atgaccaacc tccaggcggc tcagggccac gcctcctgcc
     2881 tgagccctgc tacaccctgt tgcgcgaaga tgcactgcga gccgctgacc gcagacgagg
     2941 ctcacgtgca tctccaggaa gagggcgcct tcggccacga tccgcatctg catgggtgct
     3001 tgcagggccg tactaagccg ccctacttct cccgccatcc cgccgaggaa cccgaatacg
     3061 cataccacat tcccagttcc ttggacggca gtccaagatc cagccgcacg gactcaccac
     3121 cgagctctct ggacggccca gccggtactc acaccttggc ctgttgcacc cagggagacc
     3181 cttttcccat ggtgacccag cctgagggct ctgacggttc accggcactg tattcctgcc
     3241 caacccagcc cggtagggag gcagccctcg ggccaggcca cctggaaatg ctgcgcagga
     3301 ctcagagtct tcccttcggc ggtccctccc agaatggtct gccaaaaggg aagttgctgg
     3361 aaggcctgcc atttggcacc gatggcaccg gaaacattcg cacgggacct tggaagaacg
     3421 agacgactgt gggttctgac tacaaagacg atgacgacaa gtgaagatct gacaaaactc
     3481 acacatgccc accgtgccca gcacctgaac tcctgggggg accgtcagtc ttcctcttcc
     3541 ccccaaaacc caaggacacc ctcatgatct cccggacccc tgaggtcaca tgcgtggtgg
     3601 tggacgtgag ccacgaagac cctgaggtca agttcaactg gtacgtggac ggcgtggagg
     3661 tgcataatgc caagacaaag ccgcgggagg agcagtacaa cagcacgtac cgtgtggtca
     3721 gcgtcctcac cgtcctgcac caggactggc tgaatggcaa ggagtacaag tgcaaggtct
     3781 ccaacaaagc cctcccagcc cccatcgaga aaaccatctc caaagccaaa gggcagcccc
     3841 gagaaccaca ggtgtacacc ctgcccccat cccgggagga gatgaccaag aaccaggtca
     3901 gcctgacctg cctggtcaaa ggcttctatc ccagcgacat cgccgtggag tgggagagca
     3961 atgggcagcc ggagaacaac tacaagacca cgcctcccgt gctggactcc gacggctcct
     4021 tcttcctcta cagcaagctc accgtggaca agagcaggtg gcagcagggg aacgtcttct
     4081 catgctccgt gatgcacgag gctctgcaca accactacac gcagaagagc ctctccctgt
     4141 ctccgggtaa atgagtgcta gctggccaga catgataaga tacattgatg agtttggaca
     4201 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     4261 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
     4321 tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa
     4381 atgtggtatg gaattaattc taaaatacag catagcaaaa ctttaacctc caaatcaagc
     4441 ctctacttga atccttttct gagggatgaa taaggcatag gcatcagggg ctgttgccaa
     4501 tgtgcattag ctgtttgcag cctcaccttc tttcatggag tttaagatat agtgtatttt
     4561 cccaaggttt gaactagctc ttcatttctt tatgttttaa atgcactgac ctcccacatt
     4621 ccctttttag taaaatattc agaaataatt taaatacatc attgcaatga aaataaatgt
     4681 tttttattag gcagaatcca gatgctcaag gcccttcata atatccccca gtttagtagt
     4741 tggacttagg gaacaaagga acctttaata gaaattggac agcaagaaag cgagcttcta
     4801 gcttatcctc agtcctgctc ctctgccaca aagtgcacgc agttgccggc cgggtcgcgc
     4861 agggcgaact cccgccccca cggctgctcg ccgatctcgg tcatggccgg cccggaggcg
     4921 tcccggaagt tcgtggacac gacctccgac cactcggcgt acagctcgtc caggccgcgc
     4981 acccacaccc aggccagggt gttgtccggc accacctggt cctggaccgc gctgatgaac
     5041 agggtcacgt cgtcccggac cacaccggcg aagtcgtcct ccacgaagtc ccgggagaac
     5101 ccgagccggt cggtccagaa ctcgaccgct ccggcgacgt cgcgcgcggt gagcaccgga
     5161 acggcactgg tcaacttggc catgatggct cctcctgtca ggagaggaaa gagaagaagg
     5221 ttagtacaat tgctatagtg agttgtatta tactatgcag atatactatg ccaatgatta
     5281 attgtcaaac tagggctgca gggttcatag tgccactttt cctgcactgc cccatctcct
     5341 gcccaccctt tcccaggcat agacagtcag tgacttacca aactcacagg agggagaagg
     5401 cagaagcttg agacagaccc gcgggaccgc cgaactgcga ggggacgtgg ctagggcggc
     5461 ttcttttatg gtgcgccggc cctcggaggc agggcgctcg gggaggccta gcggccaatc
     5521 tgcggtggca ggaggcgggg ccgaaggccg tgcctgacca atccggagca cataggagtc
     5581 tcagcccccc gccccaaagc aaggggaagt cacgcgcctg tagcgccagc gtgttgtgaa
     5641 atgggggctt gggggggttg gggccctgac tagtcaaaac aaactcccat tgacgtcaat
     5701 ggggtggaga cttggaaatc cccgtgagtc aaaccgctat ccacgcccat tgatgtactg
     5761 ccaaaaccgc atcatcatgg taatagcgat gactaatacg tagatgtact gccaagtagg
     5821 aaagtcccat aaggtcatgt actgggcata atgccaggcg ggccatttac cgtcattgac
     5881 gtcaataggg ggcgtacttg gcatatgata cacttgatgt actgccaagt gggcagttta
     5941 ccgtaaatac tccacccatt gacgtcaatg gaaagtccct attggcgtta ctatgggaac
     6001 atacgtcatt attgacgtca atgggcgggg gtcgttgggc ggtcagccag gcgggccatt
     6061 taccgtaagt tatgtaacgc ctgcaggtta attaagaaca tgtgagcaaa aggccagcaa
     6121 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     6181 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     6241 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     6301 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     6361 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     6421 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     6481 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     6541 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     6601 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     6661 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     6721 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     6781 gctcagtgga acgaaaactc acgttaaggg attttggtca tggctagtta attaacattt
     6841 aaatcagcgg ccgcaataaa atatctttat tttcattaca tctgtgtgtt ggttttttgt
     6901 gtgaatcgta actaacatac gctctccatc aaaacaaaac gaaacaaaac aaactagcaa
     6961 aataggctgt ccccagtgca agtgcaggtg cca
//
