LOCUS       sequence-430140-        6276 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 6276)
  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 6276)
  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-ADGRG1 CTFdeltaTA-FLAG
FEATURES             Location/Qualifiers
     source          1..6276
                     /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"
     regulatory      2403..2412
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2721..2744
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    3459..3580
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3470..3489)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3524..3543
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     polyA_signal    3684..4078
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     CDS             complement(4092..4466)
                     /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        5059..5362
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(5442..6030)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5522..5541)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    6138..6186
                     /note="synthetic polyadenylation signal"
     misc_feature    join(6197..6276,1..12)
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     6237..6256
                     /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 aagtctcctc ggtggaggtg gacgccgtgc acaagcacta cctgagcctc ctctcctacg
     1861 tgggctgtgt cgtctctgcc ctggcctgcc ttgtcaccat tgccgcctac ctctgctcca
     1921 gggtgcccct gccgtgcagg aggaaacctc gggactacac catcaaggtg cacatgaacc
     1981 tgctgctggc cgtcttcctg ctggacacga gcttcctgct cagcgagccg gtggccctga
     2041 caggctctga ggctggctgc cgagccagtg ccatcttcct gcacttctcc ctgctcacct
     2101 gcctttcctg gatgggcctc gaggggtaca acctctaccg actcgtggtg gaggtctttg
     2161 gcacctatgt ccctggctac ctactcaagc tgagcgccat gggctggggc ttccccatct
     2221 ttctggtgac gctggtggcc ctggtggatg tggacaacta tggccccatc atcttggctg
     2281 tgcataggac tccagagggc gtcatctacc cttccatgtg ctggatccgg gactccctgg
     2341 tcagctacat caccaacctg ggcctcttca gcctggtgtt tctgttcaac atggccatgc
     2401 tagccaccat ggtggtgcag atcctgcggc tgcgccccca cacccaaaag tggtcacatg
     2461 tgctgacact gctgggcctc agcctggtcc ttggcctgcc ctgggccttg atcttcttct
     2521 cctttgcttc tggcaccttc cagcttgtcg tcctctacct tttcagcatc atcacctcct
     2581 tccaaggctt cctcatcttc atctggtact ggtccatgcg gctgcaggcc cggggtggcc
     2641 cctcccctct gaagagcaac tcagacagcg ccaggctccc catcagctcg ggcagcacct
     2701 cgtccagccg catcggttct gactacaaag acgatgacga caagtgaaga tctgacaaaa
     2761 ctcacacatg cccaccgtgc ccagcacctg aactcctggg gggaccgtca gtcttcctct
     2821 tccccccaaa acccaaggac accctcatga tctcccggac ccctgaggtc acatgcgtgg
     2881 tggtggacgt gagccacgaa gaccctgagg tcaagttcaa ctggtacgtg gacggcgtgg
     2941 aggtgcataa tgccaagaca aagccgcggg aggagcagta caacagcacg taccgtgtgg
     3001 tcagcgtcct caccgtcctg caccaggact ggctgaatgg caaggagtac aagtgcaagg
     3061 tctccaacaa agccctccca gcccccatcg agaaaaccat ctccaaagcc aaagggcagc
     3121 cccgagaacc acaggtgtac accctgcccc catcccggga ggagatgacc aagaaccagg
     3181 tcagcctgac ctgcctggtc aaaggcttct atcccagcga catcgccgtg gagtgggaga
     3241 gcaatgggca gccggagaac aactacaaga ccacgcctcc cgtgctggac tccgacggct
     3301 ccttcttcct ctacagcaag ctcaccgtgg acaagagcag gtggcagcag gggaacgtct
     3361 tctcatgctc cgtgatgcac gaggctctgc acaaccacta cacgcagaag agcctctccc
     3421 tgtctccggg taaatgagtg ctagctggcc agacatgata agatacattg atgagtttgg
     3481 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     3541 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     3601 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
     3661 caaatgtggt atggaattaa ttctaaaata cagcatagca aaactttaac ctccaaatca
     3721 agcctctact tgaatccttt tctgagggat gaataaggca taggcatcag gggctgttgc
     3781 caatgtgcat tagctgtttg cagcctcacc ttctttcatg gagtttaaga tatagtgtat
     3841 tttcccaagg tttgaactag ctcttcattt ctttatgttt taaatgcact gacctcccac
     3901 attccctttt tagtaaaata ttcagaaata atttaaatac atcattgcaa tgaaaataaa
     3961 tgttttttat taggcagaat ccagatgctc aaggcccttc ataatatccc ccagtttagt
     4021 agttggactt agggaacaaa ggaaccttta atagaaattg gacagcaaga aagcgagctt
     4081 ctagcttatc ctcagtcctg ctcctctgcc acaaagtgca cgcagttgcc ggccgggtcg
     4141 cgcagggcga actcccgccc ccacggctgc tcgccgatct cggtcatggc cggcccggag
     4201 gcgtcccgga agttcgtgga cacgacctcc gaccactcgg cgtacagctc gtccaggccg
     4261 cgcacccaca cccaggccag ggtgttgtcc ggcaccacct ggtcctggac cgcgctgatg
     4321 aacagggtca cgtcgtcccg gaccacaccg gcgaagtcgt cctccacgaa gtcccgggag
     4381 aacccgagcc ggtcggtcca gaactcgacc gctccggcga cgtcgcgcgc ggtgagcacc
     4441 ggaacggcac tggtcaactt ggccatgatg gctcctcctg tcaggagagg aaagagaaga
     4501 aggttagtac aattgctata gtgagttgta ttatactatg cagatatact atgccaatga
     4561 ttaattgtca aactagggct gcagggttca tagtgccact tttcctgcac tgccccatct
     4621 cctgcccacc ctttcccagg catagacagt cagtgactta ccaaactcac aggagggaga
     4681 aggcagaagc ttgagacaga cccgcgggac cgccgaactg cgaggggacg tggctagggc
     4741 ggcttctttt atggtgcgcc ggccctcgga ggcagggcgc tcggggaggc ctagcggcca
     4801 atctgcggtg gcaggaggcg gggccgaagg ccgtgcctga ccaatccgga gcacatagga
     4861 gtctcagccc cccgccccaa agcaagggga agtcacgcgc ctgtagcgcc agcgtgttgt
     4921 gaaatggggg cttggggggg ttggggccct gactagtcaa aacaaactcc cattgacgtc
     4981 aatggggtgg agacttggaa atccccgtga gtcaaaccgc tatccacgcc cattgatgta
     5041 ctgccaaaac cgcatcatca tggtaatagc gatgactaat acgtagatgt actgccaagt
     5101 aggaaagtcc cataaggtca tgtactgggc ataatgccag gcgggccatt taccgtcatt
     5161 gacgtcaata gggggcgtac ttggcatatg atacacttga tgtactgcca agtgggcagt
     5221 ttaccgtaaa tactccaccc attgacgtca atggaaagtc cctattggcg ttactatggg
     5281 aacatacgtc attattgacg tcaatgggcg ggggtcgttg ggcggtcagc caggcgggcc
     5341 atttaccgta agttatgtaa cgcctgcagg ttaattaaga acatgtgagc aaaaggccag
     5401 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     5461 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     5521 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     5581 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     5641 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     5701 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     5761 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     5821 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     5881 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     5941 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     6001 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     6061 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatggctag ttaattaaca
     6121 tttaaatcag cggccgcaat aaaatatctt tattttcatt acatctgtgt gttggttttt
     6181 tgtgtgaatc gtaactaaca tacgctctcc atcaaaacaa aacgaaacaa aacaaactag
     6241 caaaataggc tgtccccagt gcaagtgcag gtgcca
//
