LOCUS       sequence-430277-        6354 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 6354)
  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 6354)
  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-ADGRD1 CTF-FLAG
FEATURES             Location/Qualifiers
     source          1..6354
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    149..543
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     CDS             complement(557..931)
                     /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        1524..1827
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(1907..2495)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1987..2006)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    2603..2651
                     /note="synthetic polyadenylation signal"
     misc_feature    2662..2753
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     2702..2721
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        2790..3001
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             3014..3282
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     sig_peptide     3332..3391
                     /codon_start=1
                     /product="human interleukin-2 signal sequence"
                     /label=IL-2 signal sequence
                     /translation="MYRMQLLSCIALSLALVTNS"
     CDS             3392..3418
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             5540..5563
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    join(6278..6354,1..45)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6289..6308)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     join(6343..6354,1..8)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
       61 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac
      121 ctctacaaat gtggtatgga attaattcta aaatacagca tagcaaaact ttaacctcca
      181 aatcaagcct ctacttgaat ccttttctga gggatgaata aggcataggc atcaggggct
      241 gttgccaatg tgcattagct gtttgcagcc tcaccttctt tcatggagtt taagatatag
      301 tgtattttcc caaggtttga actagctctt catttcttta tgttttaaat gcactgacct
      361 cccacattcc ctttttagta aaatattcag aaataattta aatacatcat tgcaatgaaa
      421 ataaatgttt tttattaggc agaatccaga tgctcaaggc ccttcataat atcccccagt
      481 ttagtagttg gacttaggga acaaaggaac ctttaataga aattggacag caagaaagcg
      541 agcttctagc ttatcctcag tcctgctcct ctgccacaaa gtgcacgcag ttgccggccg
      601 ggtcgcgcag ggcgaactcc cgcccccacg gctgctcgcc gatctcggtc atggccggcc
      661 cggaggcgtc ccggaagttc gtggacacga cctccgacca ctcggcgtac agctcgtcca
      721 ggccgcgcac ccacacccag gccagggtgt tgtccggcac cacctggtcc tggaccgcgc
      781 tgatgaacag ggtcacgtcg tcccggacca caccggcgaa gtcgtcctcc acgaagtccc
      841 gggagaaccc gagccggtcg gtccagaact cgaccgctcc ggcgacgtcg cgcgcggtga
      901 gcaccggaac ggcactggtc aacttggcca tgatggctcc tcctgtcagg agaggaaaga
      961 gaagaaggtt agtacaattg ctatagtgag ttgtattata ctatgcagat atactatgcc
     1021 aatgattaat tgtcaaacta gggctgcagg gttcatagtg ccacttttcc tgcactgccc
     1081 catctcctgc ccaccctttc ccaggcatag acagtcagtg acttaccaaa ctcacaggag
     1141 ggagaaggca gaagcttgag acagacccgc gggaccgccg aactgcgagg ggacgtggct
     1201 agggcggctt cttttatggt gcgccggccc tcggaggcag ggcgctcggg gaggcctagc
     1261 ggccaatctg cggtggcagg aggcggggcc gaaggccgtg cctgaccaat ccggagcaca
     1321 taggagtctc agccccccgc cccaaagcaa ggggaagtca cgcgcctgta gcgccagcgt
     1381 gttgtgaaat gggggcttgg gggggttggg gccctgacta gtcaaaacaa actcccattg
     1441 acgtcaatgg ggtggagact tggaaatccc cgtgagtcaa accgctatcc acgcccattg
     1501 atgtactgcc aaaaccgcat catcatggta atagcgatga ctaatacgta gatgtactgc
     1561 caagtaggaa agtcccataa ggtcatgtac tgggcataat gccaggcggg ccatttaccg
     1621 tcattgacgt caataggggg cgtacttggc atatgataca cttgatgtac tgccaagtgg
     1681 gcagtttacc gtaaatactc cacccattga cgtcaatgga aagtccctat tggcgttact
     1741 atgggaacat acgtcattat tgacgtcaat gggcgggggt cgttgggcgg tcagccaggc
     1801 gggccattta ccgtaagtta tgtaacgcct gcaggttaat taagaacatg tgagcaaaag
     1861 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     1921 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     1981 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     2041 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     2101 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     2161 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     2221 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     2281 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     2341 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     2401 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     2461 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     2521 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg gctagttaat
     2581 taacatttaa atcagcggcc gcaataaaat atctttattt tcattacatc tgtgtgttgg
     2641 ttttttgtgt gaatcgtaac taacatacgc tctccatcaa aacaaaacga aacaaaacaa
     2701 actagcaaaa taggctgtcc ccagtgcaag tgcaggtgcc agaacatttc tctatcgaag
     2761 gatctgcgat cgctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga
     2821 gaagttgggg ggaggggtcg gcaattgaac gggtgcctag agaaggtggc gcggggtaaa
     2881 ctgggaaagt gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta
     2941 tataagtgca gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca
     3001 gctgaagctt cgaggggctc gcatctctcc ttcacgcgcc cgccgcccta cctgaggccg
     3061 ccatccacgc cggttgagtc gcgttctgcc gcctcccgcc tgtggtgcct cctgaactgc
     3121 gtccgccgtc taggtaagtt taaagctcag gtcgagaccg ggcctttgtc cggcgctccc
     3181 ttggagccta cctagactca gccggctctc cacgctttgc ctgaccctgc ttgctcaact
     3241 ctacgtcttt gtttcgtttt ctgttctgcg ccgttacaga tccaagctgt gaccggcgcc
     3301 tacctgagat caccggcgaa ggagggccac catgtacagg atgcaactcc tgtcttgcat
     3361 tgcactaagt cttgcacttg tcacgaattc gtacccatac gatgttccag attacgctgg
     3421 atccaaaatt gaagaaggca aactggtgat ttggattaac ggcgataaag gctataacgg
     3481 cctggcggaa gtgggcaaaa aatttgaaaa agataccggc attaaagtga ccgtggaaca
     3541 tccggataaa ctggaagaaa aatttccgca ggtggcggcg accggcgatg gcccggatat
     3601 tattttttgg gcgcatgatc gctttggcgg ctatgcgcag agcggcctgc tggcggaaat
     3661 taccccggat aaagcgtttc aggataaact gtatccgttt acctgggatg cggtgcgcta
     3721 taacggcaaa ctgattgcgt atccgattgc ggtggaagcg ctgagcctga tttataacaa
     3781 agatctgctg ccgaacccgc cgaaaacctg ggaagaaatt ccggcgctgg ataaagaact
     3841 gaaagcgaaa ggcaaaagcg cgctgatgtt taacctgcag gaaccgtatt ttacctggcc
     3901 gctgattgcg gcggatggcg gctatgcgtt taaatatgaa aacggcaaat atgatattaa
     3961 agatgtgggc gtggataacg cgggcgcgaa agcgggcctg acctttctgg tggatctgat
     4021 taaaaacaaa catatgaacg cggataccga ttatagcatt gcggaagcgg cgtttaacaa
     4081 aggcgaaacc gcgatgacca ttaacggccc gtgggcgtgg agcaacattg ataccagcaa
     4141 agtgaactat ggcgtgaccg tgctgccgac ctttaaaggc cagccgagca aaccgtttgt
     4201 gggcgtgctg agcgcgggca ttaacgcggc gagcccgaac aaagaactgg cgaaagaatt
     4261 tctggaaaac tatctgctga ccgatgaagg cctggaagcg gtgaacaaag ataaaccgct
     4321 gggcgcggtg gcgctgaaaa gctatgaaga agaactggcg aaagatccgc gcattgcggc
     4381 gaccatggaa aacgcgcaga aaggcgaaat tatgccgaac attccgcaga tgagcgcgtt
     4441 ttggtatgcg gtgcgcaccg cggtgattaa cgcggcgagc ggccgccaga ccgtggatga
     4501 agcgctgaaa gatgcgcaga ccaacgagaa cctttacttc caaactaatt tcgctattct
     4561 tatgcaggtg gtccctctgg agctcgccag agggcaccaa gtggccctgt ccagcatctc
     4621 ctatgtgggt tgcagtctca gtgtcctgtg cctggtcgct accttggtta cattcgcggt
     4681 cctgtccagc gtgtccacca tccggaacca gcgctatcac atccacgcca acctgtcttt
     4741 cgccgtgctg gtcgcccagg tgctcctgct catttccttt cgtctggagc ctggaaccac
     4801 tccatgccag gtgatggctg tgcttctgca ctacttcttt ctcagcgcct tcgcctggat
     4861 gctggtggag ggcttgcacc tgtactctat ggtgatcaag gtgtttggct ctgaagactc
     4921 aaagcatcgt tactattacg gtatgggttg gggattcccc cttttgattt gcattatcag
     4981 tctgagcttc gctatggact cctacggcac ttcaaataac tgctggctgt ccctggcatc
     5041 tggcgccatc tgggcctttg tggcccccgc tctgttcgtg atcgtggtca atatcggcat
     5101 cctcattgcg gtcacaaggg tcatcagcca gatctctgca gataactaca agatccacgg
     5161 ggacccgtcc gccttcaagc tgaccgcaaa ggctgtcgcc gtcctcctgc cgattttggg
     5221 cacgtcttgg gttttcggcg ttctcgctgt caacggatgt gctgtcgtgt tccagtatat
     5281 gttcgctacc ctgaatagcc tgcaggggct cttcattttc ctgttccact gcctgctcaa
     5341 tagtgaggtg agggcggctt tcaagcacaa gaccaaggtg tggagcctga catcatcttc
     5401 cgcccggact tctaacgcga aaccgtttca tagcgacctg atgaacggga cccgcccggg
     5461 gatggcttcc accaagctct ctccctggga caagagctct catagcgccc acagagtgga
     5521 cctgtccgcc gtaggttctg actacaaaga cgatgacgac aagtgaagat ctgacaaaac
     5581 tcacacatgc ccaccgtgcc cagcacctga actcctgggg ggaccgtcag tcttcctctt
     5641 ccccccaaaa cccaaggaca ccctcatgat ctcccggacc cctgaggtca catgcgtggt
     5701 ggtggacgtg agccacgaag accctgaggt caagttcaac tggtacgtgg acggcgtgga
     5761 ggtgcataat gccaagacaa agccgcggga ggagcagtac aacagcacgt accgtgtggt
     5821 cagcgtcctc accgtcctgc accaggactg gctgaatggc aaggagtaca agtgcaaggt
     5881 ctccaacaaa gccctcccag cccccatcga gaaaaccatc tccaaagcca aagggcagcc
     5941 ccgagaacca caggtgtaca ccctgccccc atcccgggag gagatgacca agaaccaggt
     6001 cagcctgacc tgcctggtca aaggcttcta tcccagcgac atcgccgtgg agtgggagag
     6061 caatgggcag ccggagaaca actacaagac cacgcctccc gtgctggact ccgacggctc
     6121 cttcttcctc tacagcaagc tcaccgtgga caagagcagg tggcagcagg ggaacgtctt
     6181 ctcatgctcc gtgatgcacg aggctctgca caaccactac acgcagaaga gcctctccct
     6241 gtctccgggt aaatgagtgc tagctggcca gacatgataa gatacattga tgagtttgga
     6301 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgat
//
