LOCUS       sequence-429966-        6243 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 6243)
  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 6243)
  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-ADGRG3 CTFdeltaTA-FLAG
FEATURES             Location/Qualifiers
     source          1..6243
                     /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             2670..2693
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    3408..3529
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3419..3438)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3473..3492
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     polyA_signal    3633..4027
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     CDS             complement(4041..4415)
                     /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        5008..5311
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(5391..5979)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5471..5490)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    6087..6135
                     /note="synthetic polyadenylation signal"
     misc_feature    6146..6237
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     6186..6205
                     /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 ccaacgcccc actcttgatc
     1801 agtcaaccgt tcacatcctt acccgcatct ctcaggctgg ctgcggcgtg agcatgatct
     1861 tcctggcatt caccattatc ctgtatgctt tcctgcgtct gtcccgcgag cgctttaaaa
     1921 gcgaggatgc ccccaagatt cacgtggctc tgggcgggag cctgttcctg ctcaatttgg
     1981 cttttctggt taacgtggga tccggatcaa agggtagcga cgccgcttgc tgggcgaggg
     2041 gcgccgtgtt tcactacttc ctcctgtgtg ctttcacttg gatgggcttg gaggccttcc
     2101 atctgtacct cctggctgtt cgcgtgttca acacttattt cggccattat ttcctgaagc
     2161 tttccctcgt gggttggggc ctgccggccc tgatggtgat cggtaccggc tccgcaaatt
     2221 cctacggact gtacacaatc cgcgacagag aaaaccgcac gagcctggaa ctgtgttggt
     2281 tccgcgaggg cacgaccatg tacgccctgt acatcaccgt gcacggatat ttcctgatca
     2341 cattcctgtt tggaatggtc gtgctggctc ttgtggtctg gaagatcttc actctctccc
     2401 gggcgaccgc ggtcaaggag cgtggtaaga ataggaagaa agtcctgact cttctgggcc
     2461 tgagctccct ggtgggcgtg acttggggtc tggccatctt cactccactc ggcctgagca
     2521 ctgtctacat cttcgccttg tttaattcct tgcagggtgt attcatctgc tgttggttta
     2581 ctatcctcta cctgccgtcc cagtccacaa ctgtctcctc ttccaccgct agactggatc
     2641 aggcgcactc tgcttctcag gaaggttctg actacaaaga cgatgacgac aagtgaagat
     2701 ctgacaaaac tcacacatgc ccaccgtgcc cagcacctga actcctgggg ggaccgtcag
     2761 tcttcctctt ccccccaaaa cccaaggaca ccctcatgat ctcccggacc cctgaggtca
     2821 catgcgtggt ggtggacgtg agccacgaag accctgaggt caagttcaac tggtacgtgg
     2881 acggcgtgga ggtgcataat gccaagacaa agccgcggga ggagcagtac aacagcacgt
     2941 accgtgtggt cagcgtcctc accgtcctgc accaggactg gctgaatggc aaggagtaca
     3001 agtgcaaggt ctccaacaaa gccctcccag cccccatcga gaaaaccatc tccaaagcca
     3061 aagggcagcc ccgagaacca caggtgtaca ccctgccccc atcccgggag gagatgacca
     3121 agaaccaggt cagcctgacc tgcctggtca aaggcttcta tcccagcgac atcgccgtgg
     3181 agtgggagag caatgggcag ccggagaaca actacaagac cacgcctccc gtgctggact
     3241 ccgacggctc cttcttcctc tacagcaagc tcaccgtgga caagagcagg tggcagcagg
     3301 ggaacgtctt ctcatgctcc gtgatgcacg aggctctgca caaccactac acgcagaaga
     3361 gcctctccct gtctccgggt aaatgagtgc tagctggcca gacatgataa gatacattga
     3421 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     3481 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
     3541 ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta
     3601 aaacctctac aaatgtggta tggaattaat tctaaaatac agcatagcaa aactttaacc
     3661 tccaaatcaa gcctctactt gaatcctttt ctgagggatg aataaggcat aggcatcagg
     3721 ggctgttgcc aatgtgcatt agctgtttgc agcctcacct tctttcatgg agtttaagat
     3781 atagtgtatt ttcccaaggt ttgaactagc tcttcatttc tttatgtttt aaatgcactg
     3841 acctcccaca ttcccttttt agtaaaatat tcagaaataa tttaaataca tcattgcaat
     3901 gaaaataaat gttttttatt aggcagaatc cagatgctca aggcccttca taatatcccc
     3961 cagtttagta gttggactta gggaacaaag gaacctttaa tagaaattgg acagcaagaa
     4021 agcgagcttc tagcttatcc tcagtcctgc tcctctgcca caaagtgcac gcagttgccg
     4081 gccgggtcgc gcagggcgaa ctcccgcccc cacggctgct cgccgatctc ggtcatggcc
     4141 ggcccggagg cgtcccggaa gttcgtggac acgacctccg accactcggc gtacagctcg
     4201 tccaggccgc gcacccacac ccaggccagg gtgttgtccg gcaccacctg gtcctggacc
     4261 gcgctgatga acagggtcac gtcgtcccgg accacaccgg cgaagtcgtc ctccacgaag
     4321 tcccgggaga acccgagccg gtcggtccag aactcgaccg ctccggcgac gtcgcgcgcg
     4381 gtgagcaccg gaacggcact ggtcaacttg gccatgatgg ctcctcctgt caggagagga
     4441 aagagaagaa ggttagtaca attgctatag tgagttgtat tatactatgc agatatacta
     4501 tgccaatgat taattgtcaa actagggctg cagggttcat agtgccactt ttcctgcact
     4561 gccccatctc ctgcccaccc tttcccaggc atagacagtc agtgacttac caaactcaca
     4621 ggagggagaa ggcagaagct tgagacagac ccgcgggacc gccgaactgc gaggggacgt
     4681 ggctagggcg gcttctttta tggtgcgccg gccctcggag gcagggcgct cggggaggcc
     4741 tagcggccaa tctgcggtgg caggaggcgg ggccgaaggc cgtgcctgac caatccggag
     4801 cacataggag tctcagcccc ccgccccaaa gcaaggggaa gtcacgcgcc tgtagcgcca
     4861 gcgtgttgtg aaatgggggc ttgggggggt tggggccctg actagtcaaa acaaactccc
     4921 attgacgtca atggggtgga gacttggaaa tccccgtgag tcaaaccgct atccacgccc
     4981 attgatgtac tgccaaaacc gcatcatcat ggtaatagcg atgactaata cgtagatgta
     5041 ctgccaagta ggaaagtccc ataaggtcat gtactgggca taatgccagg cgggccattt
     5101 accgtcattg acgtcaatag ggggcgtact tggcatatga tacacttgat gtactgccaa
     5161 gtgggcagtt taccgtaaat actccaccca ttgacgtcaa tggaaagtcc ctattggcgt
     5221 tactatggga acatacgtca ttattgacgt caatgggcgg gggtcgttgg gcggtcagcc
     5281 aggcgggcca tttaccgtaa gttatgtaac gcctgcaggt taattaagaa catgtgagca
     5341 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     5401 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     5461 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     5521 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     5581 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     5641 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     5701 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     5761 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     5821 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     5881 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     5941 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     6001 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catggctagt
     6061 taattaacat ttaaatcagc ggccgcaata aaatatcttt attttcatta catctgtgtg
     6121 ttggtttttt gtgtgaatcg taactaacat acgctctcca tcaaaacaaa acgaaacaaa
     6181 acaaactagc aaaataggct gtccccagtg caagtgcagg tgccagaaca tttctctatc
     6241 gaa
//
