LOCUS       sequence-430126-        6216 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 6216)
  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 6216)
  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-ADGRL4 CTF-FLAG
FEATURES             Location/Qualifiers
     source          1..6216
                     /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             2661..2684
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    3399..3520
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3410..3429)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3464..3483
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     polyA_signal    3624..4018
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     CDS             complement(4032..4406)
                     /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        4999..5302
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(5382..5970)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5462..5481)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    6078..6126
                     /note="synthetic polyadenylation signal"
     misc_feature    join(6137..6216,1..12)
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     6177..6196
                     /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 aaacccactt cgctattctg atgagctccg ggcctagcat cggcatcaag gactataata
     1861 tcttgaccag gattacacag ctgggtatca ttatctcttt gatctgtttg gctatttgta
     1921 tcttcacctt ctggtttttc agtgaaatcc agtctacccg gacaaccatc cataagaatc
     1981 tttgttgctc cctgttcctg gccgagctgg tgttcctggt ggggatcaat actaacacta
     2041 acaagctgtt ctgttcaatt atcgccggct tgctgcacta ctttttcctc gcagctttcg
     2101 cctggatgtg cattgaaggt attcacctct atctgattgt tgtgggggtg atttacaaca
     2161 aaggcttcct gcacaagaac ttctacatct ttggctacct gtctcccgcc gtcgtggttg
     2221 gatttagcgc ggccctgggg tatcgttatt acggcacaac caaggtttgt tggttgagca
     2281 cagaaaataa cttcatctgg tctttcattg gtcctgcatg cctgattatc ctggtcaacc
     2341 tgcttgcctt tggcgtaatt atctacaagg ttttccgcca caccgcgggc ctgaagcccg
     2401 aagtgtcttg tttcgaaaac atccggtcct gcgctcgcgg agctctggcc ctcctgtttc
     2461 tcctgggcac cacttggatt ttcggcgtcc tgcacgtcgt gcacgcctcc gtggtcacag
     2521 cctacctgtt caccgtgtct aacgcattcc agggcatgtt catcttcctg ttcctgtgtg
     2581 tactgtcccg caagatccaa gaagagtatt accgtctgtt caagaacgtg ccatgctgtt
     2641 ttggctgtct cagaggttct gactacaaag acgatgacga caagtgaaga tctgacaaaa
     2701 ctcacacatg cccaccgtgc ccagcacctg aactcctggg gggaccgtca gtcttcctct
     2761 tccccccaaa acccaaggac accctcatga tctcccggac ccctgaggtc acatgcgtgg
     2821 tggtggacgt gagccacgaa gaccctgagg tcaagttcaa ctggtacgtg gacggcgtgg
     2881 aggtgcataa tgccaagaca aagccgcggg aggagcagta caacagcacg taccgtgtgg
     2941 tcagcgtcct caccgtcctg caccaggact ggctgaatgg caaggagtac aagtgcaagg
     3001 tctccaacaa agccctccca gcccccatcg agaaaaccat ctccaaagcc aaagggcagc
     3061 cccgagaacc acaggtgtac accctgcccc catcccggga ggagatgacc aagaaccagg
     3121 tcagcctgac ctgcctggtc aaaggcttct atcccagcga catcgccgtg gagtgggaga
     3181 gcaatgggca gccggagaac aactacaaga ccacgcctcc cgtgctggac tccgacggct
     3241 ccttcttcct ctacagcaag ctcaccgtgg acaagagcag gtggcagcag gggaacgtct
     3301 tctcatgctc cgtgatgcac gaggctctgc acaaccacta cacgcagaag agcctctccc
     3361 tgtctccggg taaatgagtg ctagctggcc agacatgata agatacattg atgagtttgg
     3421 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     3481 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     3541 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
     3601 caaatgtggt atggaattaa ttctaaaata cagcatagca aaactttaac ctccaaatca
     3661 agcctctact tgaatccttt tctgagggat gaataaggca taggcatcag gggctgttgc
     3721 caatgtgcat tagctgtttg cagcctcacc ttctttcatg gagtttaaga tatagtgtat
     3781 tttcccaagg tttgaactag ctcttcattt ctttatgttt taaatgcact gacctcccac
     3841 attccctttt tagtaaaata ttcagaaata atttaaatac atcattgcaa tgaaaataaa
     3901 tgttttttat taggcagaat ccagatgctc aaggcccttc ataatatccc ccagtttagt
     3961 agttggactt agggaacaaa ggaaccttta atagaaattg gacagcaaga aagcgagctt
     4021 ctagcttatc ctcagtcctg ctcctctgcc acaaagtgca cgcagttgcc ggccgggtcg
     4081 cgcagggcga actcccgccc ccacggctgc tcgccgatct cggtcatggc cggcccggag
     4141 gcgtcccgga agttcgtgga cacgacctcc gaccactcgg cgtacagctc gtccaggccg
     4201 cgcacccaca cccaggccag ggtgttgtcc ggcaccacct ggtcctggac cgcgctgatg
     4261 aacagggtca cgtcgtcccg gaccacaccg gcgaagtcgt cctccacgaa gtcccgggag
     4321 aacccgagcc ggtcggtcca gaactcgacc gctccggcga cgtcgcgcgc ggtgagcacc
     4381 ggaacggcac tggtcaactt ggccatgatg gctcctcctg tcaggagagg aaagagaaga
     4441 aggttagtac aattgctata gtgagttgta ttatactatg cagatatact atgccaatga
     4501 ttaattgtca aactagggct gcagggttca tagtgccact tttcctgcac tgccccatct
     4561 cctgcccacc ctttcccagg catagacagt cagtgactta ccaaactcac aggagggaga
     4621 aggcagaagc ttgagacaga cccgcgggac cgccgaactg cgaggggacg tggctagggc
     4681 ggcttctttt atggtgcgcc ggccctcgga ggcagggcgc tcggggaggc ctagcggcca
     4741 atctgcggtg gcaggaggcg gggccgaagg ccgtgcctga ccaatccgga gcacatagga
     4801 gtctcagccc cccgccccaa agcaagggga agtcacgcgc ctgtagcgcc agcgtgttgt
     4861 gaaatggggg cttggggggg ttggggccct gactagtcaa aacaaactcc cattgacgtc
     4921 aatggggtgg agacttggaa atccccgtga gtcaaaccgc tatccacgcc cattgatgta
     4981 ctgccaaaac cgcatcatca tggtaatagc gatgactaat acgtagatgt actgccaagt
     5041 aggaaagtcc cataaggtca tgtactgggc ataatgccag gcgggccatt taccgtcatt
     5101 gacgtcaata gggggcgtac ttggcatatg atacacttga tgtactgcca agtgggcagt
     5161 ttaccgtaaa tactccaccc attgacgtca atggaaagtc cctattggcg ttactatggg
     5221 aacatacgtc attattgacg tcaatgggcg ggggtcgttg ggcggtcagc caggcgggcc
     5281 atttaccgta agttatgtaa cgcctgcagg ttaattaaga acatgtgagc aaaaggccag
     5341 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     5401 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     5461 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     5521 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     5581 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     5641 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     5701 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     5761 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     5821 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     5881 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     5941 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     6001 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatggctag ttaattaaca
     6061 tttaaatcag cggccgcaat aaaatatctt tattttcatt acatctgtgt gttggttttt
     6121 tgtgtgaatc gtaactaaca tacgctctcc atcaaaacaa aacgaaacaa aacaaactag
     6181 caaaataggc tgtccccagt gcaagtgcag gtgcca
//
