LOCUS       sequence-430282-        6279 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 6279)
  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 6279)
  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-ADGRE5 CTF-FLAG
FEATURES             Location/Qualifiers
     source          1..6279
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    15..136
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(26..45)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     80..99
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     polyA_signal    240..634
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     CDS             complement(648..1022)
                     /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        1615..1918
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(1998..2586)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2078..2097)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    2694..2742
                     /note="synthetic polyadenylation signal"
     misc_feature    2753..2844
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     2793..2812
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        2881..3092
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             3105..3373
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     sig_peptide     3423..3482
                     /codon_start=1
                     /product="human interleukin-2 signal sequence"
                     /label=IL-2 signal sequence
                     /translation="MYRMQLLSCIALSLALVTNS"
     CDS             3483..3509
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4617..4637
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             5556..5579
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
ORIGIN
        1 ctggccagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
       61 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
      121 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga
      181 ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtatgg aattaattct
      241 aaaatacagc atagcaaaac tttaacctcc aaatcaagcc tctacttgaa tccttttctg
      301 agggatgaat aaggcatagg catcaggggc tgttgccaat gtgcattagc tgtttgcagc
      361 ctcaccttct ttcatggagt ttaagatata gtgtattttc ccaaggtttg aactagctct
      421 tcatttcttt atgttttaaa tgcactgacc tcccacattc cctttttagt aaaatattca
      481 gaaataattt aaatacatca ttgcaatgaa aataaatgtt ttttattagg cagaatccag
      541 atgctcaagg cccttcataa tatcccccag tttagtagtt ggacttaggg aacaaaggaa
      601 cctttaatag aaattggaca gcaagaaagc gagcttctag cttatcctca gtcctgctcc
      661 tctgccacaa agtgcacgca gttgccggcc gggtcgcgca gggcgaactc ccgcccccac
      721 ggctgctcgc cgatctcggt catggccggc ccggaggcgt cccggaagtt cgtggacacg
      781 acctccgacc actcggcgta cagctcgtcc aggccgcgca cccacaccca ggccagggtg
      841 ttgtccggca ccacctggtc ctggaccgcg ctgatgaaca gggtcacgtc gtcccggacc
      901 acaccggcga agtcgtcctc cacgaagtcc cgggagaacc cgagccggtc ggtccagaac
      961 tcgaccgctc cggcgacgtc gcgcgcggtg agcaccggaa cggcactggt caacttggcc
     1021 atgatggctc ctcctgtcag gagaggaaag agaagaaggt tagtacaatt gctatagtga
     1081 gttgtattat actatgcaga tatactatgc caatgattaa ttgtcaaact agggctgcag
     1141 ggttcatagt gccacttttc ctgcactgcc ccatctcctg cccacccttt cccaggcata
     1201 gacagtcagt gacttaccaa actcacagga gggagaaggc agaagcttga gacagacccg
     1261 cgggaccgcc gaactgcgag gggacgtggc tagggcggct tcttttatgg tgcgccggcc
     1321 ctcggaggca gggcgctcgg ggaggcctag cggccaatct gcggtggcag gaggcggggc
     1381 cgaaggccgt gcctgaccaa tccggagcac ataggagtct cagccccccg ccccaaagca
     1441 aggggaagtc acgcgcctgt agcgccagcg tgttgtgaaa tgggggcttg ggggggttgg
     1501 ggccctgact agtcaaaaca aactcccatt gacgtcaatg gggtggagac ttggaaatcc
     1561 ccgtgagtca aaccgctatc cacgcccatt gatgtactgc caaaaccgca tcatcatggt
     1621 aatagcgatg actaatacgt agatgtactg ccaagtagga aagtcccata aggtcatgta
     1681 ctgggcataa tgccaggcgg gccatttacc gtcattgacg tcaatagggg gcgtacttgg
     1741 catatgatac acttgatgta ctgccaagtg ggcagtttac cgtaaatact ccacccattg
     1801 acgtcaatgg aaagtcccta ttggcgttac tatgggaaca tacgtcatta ttgacgtcaa
     1861 tgggcggggg tcgttgggcg gtcagccagg cgggccattt accgtaagtt atgtaacgcc
     1921 tgcaggttaa ttaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     1981 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     2041 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     2101 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     2161 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     2221 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     2281 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     2341 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     2401 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     2461 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     2521 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     2581 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     2641 cgttaaggga ttttggtcat ggctagttaa ttaacattta aatcagcggc cgcaataaaa
     2701 tatctttatt ttcattacat ctgtgtgttg gttttttgtg tgaatcgtaa ctaacatacg
     2761 ctctccatca aaacaaaacg aaacaaaaca aactagcaaa ataggctgtc cccagtgcaa
     2821 gtgcaggtgc cagaacattt ctctatcgaa ggatctgcga tcgctccggt gcccgtcagt
     2881 gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc ggcaattgaa
     2941 cgggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg tactggctcc
     3001 gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc
     3061 tttttcgcaa cgggtttgcc gccagaacac agctgaagct tcgaggggct cgcatctctc
     3121 cttcacgcgc ccgccgccct acctgaggcc gccatccacg ccggttgagt cgcgttctgc
     3181 cgcctcccgc ctgtggtgcc tcctgaactg cgtccgccgt ctaggtaagt ttaaagctca
     3241 ggtcgagacc gggcctttgt ccggcgctcc cttggagcct acctagactc agccggctct
     3301 ccacgctttg cctgaccctg cttgctcaac tctacgtctt tgtttcgttt tctgttctgc
     3361 gccgttacag atccaagctg tgaccggcgc ctacctgaga tcaccggcga aggagggcca
     3421 ccatgtacag gatgcaactc ctgtcttgca ttgcactaag tcttgcactt gtcacgaatt
     3481 cgtacccata cgatgttcca gattacgctg gatccaaaat tgaagaaggc aaactggtga
     3541 tttggattaa cggcgataaa ggctataacg gcctggcgga agtgggcaaa aaatttgaaa
     3601 aagataccgg cattaaagtg accgtggaac atccggataa actggaagaa aaatttccgc
     3661 aggtggcggc gaccggcgat ggcccggata ttattttttg ggcgcatgat cgctttggcg
     3721 gctatgcgca gagcggcctg ctggcggaaa ttaccccgga taaagcgttt caggataaac
     3781 tgtatccgtt tacctgggat gcggtgcgct ataacggcaa actgattgcg tatccgattg
     3841 cggtggaagc gctgagcctg atttataaca aagatctgct gccgaacccg ccgaaaacct
     3901 gggaagaaat tccggcgctg gataaagaac tgaaagcgaa aggcaaaagc gcgctgatgt
     3961 ttaacctgca ggaaccgtat tttacctggc cgctgattgc ggcggatggc ggctatgcgt
     4021 ttaaatatga aaacggcaaa tatgatatta aagatgtggg cgtggataac gcgggcgcga
     4081 aagcgggcct gacctttctg gtggatctga ttaaaaacaa acatatgaac gcggataccg
     4141 attatagcat tgcggaagcg gcgtttaaca aaggcgaaac cgcgatgacc attaacggcc
     4201 cgtgggcgtg gagcaacatt gataccagca aagtgaacta tggcgtgacc gtgctgccga
     4261 cctttaaagg ccagccgagc aaaccgtttg tgggcgtgct gagcgcgggc attaacgcgg
     4321 cgagcccgaa caaagaactg gcgaaagaat ttctggaaaa ctatctgctg accgatgaag
     4381 gcctggaagc ggtgaacaaa gataaaccgc tgggcgcggt ggcgctgaaa agctatgaag
     4441 aagaactggc gaaagatccg cgcattgcgg cgaccatgga aaacgcgcag aaaggcgaaa
     4501 ttatgccgaa cattccgcag atgagcgcgt tttggtatgc ggtgcgcacc gcggtgatta
     4561 acgcggcgag cggccgccag accgtggatg aagcgctgaa agatgcgcag accaacgaga
     4621 acctttactt ccaaagcagc tttgcgatcc ttatggctca ttatgacgtg gaggactgga
     4681 agctgaccct gatcaccagg gtgggactgg cgctgtcact cttctgcctg ctgctgtgca
     4741 tcctcacttt cctgctggtg cggcccatcc agggctcgcg caccaccata cacctgcacc
     4801 tctgcatctg cctcttcgtg ggctccacca tcttcctggc cggcatcgag aacgaaggcg
     4861 gccaggtggg gctgcgctgc cgcctggtgg ccgggctgct gcactactgt ttcctggccg
     4921 ccttctgctg gatgagcctc gaaggcctgg agctctactt tcttgtggtg cgcgtgttcc
     4981 aaggccaggg cctgagtacg cgctggctct gcctgatcgg ctatggcgtg cccctgctca
     5041 tcgtgggcgt ctcggctgcc atctacagca agggctacgg ccgccccaga tactgctggt
     5101 tggactttga gcagggcttc ctctggagct tcttgggacc tgtgaccttc atcattttgt
     5161 gcaatgctgt cattttcgtg actaccgtct ggaagctcac tcagaagttt tctgaaatca
     5221 atccagacat gaagaaatta aagaaggcga gggcgctgac catcacggcc atcgcgcagc
     5281 tcttcctgtt gggctgcacc tgggtctttg gcctgttcat cttcgacgat cggagcttgg
     5341 tgctgaccta tgtgtttacc atcctcaact gcctgcaggg cgccttcctc tacctgctgc
     5401 actgcctgct caacaagaag gttcgggaag aataccggaa gtgggcctgc ctagttgctg
     5461 gggggagcaa gtactcagaa ttcacctcca ccacgtctgg cactggccac aatcagaccc
     5521 gggccctcag ggcatcagag tccggcatag gttctgacta caaagacgat gacgacaagt
     5581 gaagatctga caaaactcac acatgcccac cgtgcccagc acctgaactc ctggggggac
     5641 cgtcagtctt cctcttcccc ccaaaaccca aggacaccct catgatctcc cggacccctg
     5701 aggtcacatg cgtggtggtg gacgtgagcc acgaagaccc tgaggtcaag ttcaactggt
     5761 acgtggacgg cgtggaggtg cataatgcca agacaaagcc gcgggaggag cagtacaaca
     5821 gcacgtaccg tgtggtcagc gtcctcaccg tcctgcacca ggactggctg aatggcaagg
     5881 agtacaagtg caaggtctcc aacaaagccc tcccagcccc catcgagaaa accatctcca
     5941 aagccaaagg gcagccccga gaaccacagg tgtacaccct gcccccatcc cgggaggaga
     6001 tgaccaagaa ccaggtcagc ctgacctgcc tggtcaaagg cttctatccc agcgacatcg
     6061 ccgtggagtg ggagagcaat gggcagccgg agaacaacta caagaccacg cctcccgtgc
     6121 tggactccga cggctccttc ttcctctaca gcaagctcac cgtggacaag agcaggtggc
     6181 agcaggggaa cgtcttctca tgctccgtga tgcacgaggc tctgcacaac cactacacgc
     6241 agaagagcct ctccctgtct ccgggtaaat gagtgctag
//
