LOCUS       sequence-429943-        6261 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 6261)
  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 6261)
  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-ADGRE2 CTFdeltaTA-FLAG
FEATURES             Location/Qualifiers
     source          1..6261
                     /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             1767..1787
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQA"
     CDS             2688..2711
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    3426..3547
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3437..3456)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3491..3510
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     polyA_signal    3651..4045
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     CDS             complement(4059..4433)
                     /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        5026..5329
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(5409..5997)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5489..5508)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    6105..6153
                     /note="synthetic polyadenylation signal"
     misc_feature    6164..6255
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     6204..6223
                     /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 ccaagcccac tacgatgtgc
     1801 aggaggagga tcccgtgctg actgtcatca cctacatggg gctgagcgtc tctctgctgt
     1861 gcctcctcct ggcggccctc acttttctcc tgtgtaaagc catccagaac accagcacct
     1921 cactgcatct gcagctctcg ctctgcctct tcctggccca cctcctcttc ctcgtggcaa
     1981 ttgatcaaac cggacacaag gtgctgtgct ccatcatcgc cggtaccttg cactatctct
     2041 acctggccac cttgacctgg atgctgctgg aggccctgta cctcttcctc actgcacgga
     2101 acctgacggt ggtcaactac tcaagcatca acagattcat gaagaagctc atgttccctg
     2161 tgggctacgg agtcccagct gtgacagtgg ccatttctgc agcctccagg cctcaccttt
     2221 atggaacacc ttcccgctgc tggctccaac cagaaaaggg atttatatgg ggcttccttg
     2281 gacctgtctg cgccatcttc tctgtgaatt tagttctctt tctggtgact ctctggattt
     2341 tgaaaaacag actctcctcc ctcaatagtg aagtgtccac cctccggaac acaaggatgc
     2401 tggcatttaa agcgacagct cagctgttca tcctgggctg cacgtggtgt ctgggcatct
     2461 tgcaggtggg tccggctgcc cgggtcatgg cctacctctt caccatcatc aacagcctgc
     2521 agggtgtctt catcttcctg gtgtactgcc tcctcagcca gcaggtccgg gagcaatatg
     2581 ggaaatggtc caaagggatc aggaaattga aaactgagtc tgagatgcac acactctcca
     2641 gcagtgctaa ggctgacacc tccaaaccca gcacggttaa cggttctgac tacaaagacg
     2701 atgacgacaa gtgaagatct gacaaaactc acacatgccc accgtgccca gcacctgaac
     2761 tcctgggggg accgtcagtc ttcctcttcc ccccaaaacc caaggacacc ctcatgatct
     2821 cccggacccc tgaggtcaca tgcgtggtgg tggacgtgag ccacgaagac cctgaggtca
     2881 agttcaactg gtacgtggac ggcgtggagg tgcataatgc caagacaaag ccgcgggagg
     2941 agcagtacaa cagcacgtac cgtgtggtca gcgtcctcac cgtcctgcac caggactggc
     3001 tgaatggcaa ggagtacaag tgcaaggtct ccaacaaagc cctcccagcc cccatcgaga
     3061 aaaccatctc caaagccaaa gggcagcccc gagaaccaca ggtgtacacc ctgcccccat
     3121 cccgggagga gatgaccaag aaccaggtca gcctgacctg cctggtcaaa ggcttctatc
     3181 ccagcgacat cgccgtggag tgggagagca atgggcagcc ggagaacaac tacaagacca
     3241 cgcctcccgt gctggactcc gacggctcct tcttcctcta cagcaagctc accgtggaca
     3301 agagcaggtg gcagcagggg aacgtcttct catgctccgt gatgcacgag gctctgcaca
     3361 accactacac gcagaagagc ctctccctgt ctccgggtaa atgagtgcta gctggccaga
     3421 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     3481 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     3541 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga
     3601 ggttttttaa agcaagtaaa acctctacaa atgtggtatg gaattaattc taaaatacag
     3661 catagcaaaa ctttaacctc caaatcaagc ctctacttga atccttttct gagggatgaa
     3721 taaggcatag gcatcagggg ctgttgccaa tgtgcattag ctgtttgcag cctcaccttc
     3781 tttcatggag tttaagatat agtgtatttt cccaaggttt gaactagctc ttcatttctt
     3841 tatgttttaa atgcactgac ctcccacatt ccctttttag taaaatattc agaaataatt
     3901 taaatacatc attgcaatga aaataaatgt tttttattag gcagaatcca gatgctcaag
     3961 gcccttcata atatccccca gtttagtagt tggacttagg gaacaaagga acctttaata
     4021 gaaattggac agcaagaaag cgagcttcta gcttatcctc agtcctgctc ctctgccaca
     4081 aagtgcacgc agttgccggc cgggtcgcgc agggcgaact cccgccccca cggctgctcg
     4141 ccgatctcgg tcatggccgg cccggaggcg tcccggaagt tcgtggacac gacctccgac
     4201 cactcggcgt acagctcgtc caggccgcgc acccacaccc aggccagggt gttgtccggc
     4261 accacctggt cctggaccgc gctgatgaac agggtcacgt cgtcccggac cacaccggcg
     4321 aagtcgtcct ccacgaagtc ccgggagaac ccgagccggt cggtccagaa ctcgaccgct
     4381 ccggcgacgt cgcgcgcggt gagcaccgga acggcactgg tcaacttggc catgatggct
     4441 cctcctgtca ggagaggaaa gagaagaagg ttagtacaat tgctatagtg agttgtatta
     4501 tactatgcag atatactatg ccaatgatta attgtcaaac tagggctgca gggttcatag
     4561 tgccactttt cctgcactgc cccatctcct gcccaccctt tcccaggcat agacagtcag
     4621 tgacttacca aactcacagg agggagaagg cagaagcttg agacagaccc gcgggaccgc
     4681 cgaactgcga ggggacgtgg ctagggcggc ttcttttatg gtgcgccggc cctcggaggc
     4741 agggcgctcg gggaggccta gcggccaatc tgcggtggca ggaggcgggg ccgaaggccg
     4801 tgcctgacca atccggagca cataggagtc tcagcccccc gccccaaagc aaggggaagt
     4861 cacgcgcctg tagcgccagc gtgttgtgaa atgggggctt gggggggttg gggccctgac
     4921 tagtcaaaac aaactcccat tgacgtcaat ggggtggaga cttggaaatc cccgtgagtc
     4981 aaaccgctat ccacgcccat tgatgtactg ccaaaaccgc atcatcatgg taatagcgat
     5041 gactaatacg tagatgtact gccaagtagg aaagtcccat aaggtcatgt actgggcata
     5101 atgccaggcg ggccatttac cgtcattgac gtcaataggg ggcgtacttg gcatatgata
     5161 cacttgatgt actgccaagt gggcagttta ccgtaaatac tccacccatt gacgtcaatg
     5221 gaaagtccct attggcgtta ctatgggaac atacgtcatt attgacgtca atgggcgggg
     5281 gtcgttgggc ggtcagccag gcgggccatt taccgtaagt tatgtaacgc ctgcaggtta
     5341 attaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     5401 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     5461 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     5521 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     5581 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     5641 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     5701 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     5761 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     5821 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     5881 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     5941 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     6001 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     6061 attttggtca tggctagtta attaacattt aaatcagcgg ccgcaataaa atatctttat
     6121 tttcattaca tctgtgtgtt ggttttttgt gtgaatcgta actaacatac gctctccatc
     6181 aaaacaaaac gaaacaaaac aaactagcaa aataggctgt ccccagtgca agtgcaggtg
     6241 ccagaacatt tctctatcga a
//
