LOCUS       sequence-430009-        6333 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 6333)
  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 6333)
  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-ADGRD1 CTFdeltaTA-FLAG
FEATURES             Location/Qualifiers
     source          1..6333
                     /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             2760..2783
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    3498..3619
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3509..3528)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3563..3582
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     polyA_signal    3723..4117
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     CDS             complement(4131..4505)
                     /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        5098..5401
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(5481..6069)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5561..5580)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    6177..6225
                     /note="synthetic polyadenylation signal"
     misc_feature    6236..6327
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     6276..6295
                     /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 ccaacaggtg gtccctctgg
     1801 agctcgccag agggcaccaa gtggccctgt ccagcatctc ctatgtgggt tgcagtctca
     1861 gtgtcctgtg cctggtcgct accttggtta cattcgcggt cctgtccagc gtgtccacca
     1921 tccggaacca gcgctatcac atccacgcca acctgtcttt cgccgtgctg gtcgcccagg
     1981 tgctcctgct catttccttt cgtctggagc ctggaaccac tccatgccag gtgatggctg
     2041 tgcttctgca ctacttcttt ctcagcgcct tcgcctggat gctggtggag ggcttgcacc
     2101 tgtactctat ggtgatcaag gtgtttggct ctgaagactc aaagcatcgt tactattacg
     2161 gtatgggttg gggattcccc cttttgattt gcattatcag tctgagcttc gctatggact
     2221 cctacggcac ttcaaataac tgctggctgt ccctggcatc tggcgccatc tgggcctttg
     2281 tggcccccgc tctgttcgtg atcgtggtca atatcggcat cctcattgcg gtcacaaggg
     2341 tcatcagcca gatctctgca gataactaca agatccacgg ggacccgtcc gccttcaagc
     2401 tgaccgcaaa ggctgtcgcc gtcctcctgc cgattttggg cacgtcttgg gttttcggcg
     2461 ttctcgctgt caacggatgt gctgtcgtgt tccagtatat gttcgctacc ctgaatagcc
     2521 tgcaggggct cttcattttc ctgttccact gcctgctcaa tagtgaggtg agggcggctt
     2581 tcaagcacaa gaccaaggtg tggagcctga catcatcttc cgcccggact tctaacgcga
     2641 aaccgtttca tagcgacctg atgaacggga cccgcccggg gatggcttcc accaagctct
     2701 ctccctggga caagagctct catagcgccc acagagtgga cctgtccgcc gtaggttctg
     2761 actacaaaga cgatgacgat aagtgaagat ctgacaaaac tcacacatgc ccaccgtgcc
     2821 cagcacctga actcctgggg ggaccgtcag tcttcctctt ccccccaaaa cccaaggaca
     2881 ccctcatgat ctcccggacc cctgaggtca catgcgtggt ggtggacgtg agccacgaag
     2941 accctgaggt caagttcaac tggtacgtgg acggcgtgga ggtgcataat gccaagacaa
     3001 agccgcggga ggagcagtac aacagcacgt accgtgtggt cagcgtcctc accgtcctgc
     3061 accaggactg gctgaatggc aaggagtaca agtgcaaggt ctccaacaaa gccctcccag
     3121 cccccatcga gaaaaccatc tccaaagcca aagggcagcc ccgagaacca caggtgtaca
     3181 ccctgccccc atcccgggag gagatgacca agaaccaggt cagcctgacc tgcctggtca
     3241 aaggcttcta tcccagcgac atcgccgtgg agtgggagag caatgggcag ccggagaaca
     3301 actacaagac cacgcctccc gtgctggact ccgacggctc cttcttcctc tacagcaagc
     3361 tcaccgtgga caagagcagg tggcagcagg ggaacgtctt ctcatgctcc gtgatgcacg
     3421 aggctctgca caaccactac acgcagaaga gcctctccct gtctccgggt aaatgagtgc
     3481 tagctggcca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca
     3541 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
     3601 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg
     3661 ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta tggaattaat
     3721 tctaaaatac agcatagcaa aactttaacc tccaaatcaa gcctctactt gaatcctttt
     3781 ctgagggatg aataaggcat aggcatcagg ggctgttgcc aatgtgcatt agctgtttgc
     3841 agcctcacct tctttcatgg agtttaagat atagtgtatt ttcccaaggt ttgaactagc
     3901 tcttcatttc tttatgtttt aaatgcactg acctcccaca ttcccttttt agtaaaatat
     3961 tcagaaataa tttaaataca tcattgcaat gaaaataaat gttttttatt aggcagaatc
     4021 cagatgctca aggcccttca taatatcccc cagtttagta gttggactta gggaacaaag
     4081 gaacctttaa tagaaattgg acagcaagaa agcgagcttc tagcttatcc tcagtcctgc
     4141 tcctctgcca caaagtgcac gcagttgccg gccgggtcgc gcagggcgaa ctcccgcccc
     4201 cacggctgct cgccgatctc ggtcatggcc ggcccggagg cgtcccggaa gttcgtggac
     4261 acgacctccg accactcggc gtacagctcg tccaggccgc gcacccacac ccaggccagg
     4321 gtgttgtccg gcaccacctg gtcctggacc gcgctgatga acagggtcac gtcgtcccgg
     4381 accacaccgg cgaagtcgtc ctccacgaag tcccgggaga acccgagccg gtcggtccag
     4441 aactcgaccg ctccggcgac gtcgcgcgcg gtgagcaccg gaacggcact ggtcaacttg
     4501 gccatgatgg ctcctcctgt caggagagga aagagaagaa ggttagtaca attgctatag
     4561 tgagttgtat tatactatgc agatatacta tgccaatgat taattgtcaa actagggctg
     4621 cagggttcat agtgccactt ttcctgcact gccccatctc ctgcccaccc tttcccaggc
     4681 atagacagtc agtgacttac caaactcaca ggagggagaa ggcagaagct tgagacagac
     4741 ccgcgggacc gccgaactgc gaggggacgt ggctagggcg gcttctttta tggtgcgccg
     4801 gccctcggag gcagggcgct cggggaggcc tagcggccaa tctgcggtgg caggaggcgg
     4861 ggccgaaggc cgtgcctgac caatccggag cacataggag tctcagcccc ccgccccaaa
     4921 gcaaggggaa gtcacgcgcc tgtagcgcca gcgtgttgtg aaatgggggc ttgggggggt
     4981 tggggccctg actagtcaaa acaaactccc attgacgtca atggggtgga gacttggaaa
     5041 tccccgtgag tcaaaccgct atccacgccc attgatgtac tgccaaaacc gcatcatcat
     5101 ggtaatagcg atgactaata cgtagatgta ctgccaagta ggaaagtccc ataaggtcat
     5161 gtactgggca taatgccagg cgggccattt accgtcattg acgtcaatag ggggcgtact
     5221 tggcatatga tacacttgat gtactgccaa gtgggcagtt taccgtaaat actccaccca
     5281 ttgacgtcaa tggaaagtcc ctattggcgt tactatggga acatacgtca ttattgacgt
     5341 caatgggcgg gggtcgttgg gcggtcagcc aggcgggcca tttaccgtaa gttatgtaac
     5401 gcctgcaggt taattaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     5461 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     5521 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     5581 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     5641 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     5701 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     5761 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     5821 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     5881 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     5941 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     6001 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     6061 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     6121 tcacgttaag ggattttggt catggctagt taattaacat ttaaatcagc ggccgcaata
     6181 aaatatcttt attttcatta catctgtgtg ttggtttttt gtgtgaatcg taactaacat
     6241 acgctctcca tcaaaacaaa acgaaacaaa acaaactagc aaaataggct gtccccagtg
     6301 caagtgcagg tgccagaaca tttctctatc gaa
//
