LOCUS       sequence-442645-        6285 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 6285)
  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 6285)
  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-ADGRE3 CTFdeltaTA-FLAG
FEATURES             Location/Qualifiers
     source          1..6285
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             102..476
                     /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"
     polyA_signal    490..884
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     polyA_signal    988..1109
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1025..1044)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1079..1098
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(1824..1847)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(2772..2792)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQA"
     CDS             complement(3900..3926)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     sig_peptide     complement(3927..3986)
                     /codon_start=1
                     /product="human interleukin-2 signal sequence"
                     /label=IL-2 signal sequence
                     /translation="MYRMQLLSCIALSLALVTNS"
     LTR             4036..4304
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     promoter        complement(4317..4528)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     misc_feature    4565..4656
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     complement(4597..4616)
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     polyA_signal    4667..4715
                     /note="synthetic polyadenylation signal"
     rep_origin      4823..5411
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5312..5331
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        5491..5794
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 caattaatca ttggcatagt atatctgcat agtataatac aactcactat agcaattgta
       61 ctaaccttct tctctttcct ctcctgacag gaggagccat catggccaag ttgaccagtg
      121 ccgttccggt gctcaccgcg cgcgacgtcg ccggagcggt cgagttctgg accgaccggc
      181 tcgggttctc ccgggacttc gtggaggacg acttcgccgg tgtggtccgg gacgacgtga
      241 ccctgttcat cagcgcggtc caggaccagg tggtgccgga caacaccctg gcctgggtgt
      301 gggtgcgcgg cctggacgag ctgtacgccg agtggtcgga ggtcgtgtcc acgaacttcc
      361 gggacgcctc cgggccggcc atgaccgaga tcggcgagca gccgtggggg cgggagttcg
      421 ccctgcgcga cccggccggc aactgcgtgc actttgtggc agaggagcag gactgaggat
      481 aagctagaag ctcgctttct tgctgtccaa tttctattaa aggttccttt gttccctaag
      541 tccaactact aaactggggg atattatgaa gggccttgag catctggatt ctgcctaata
      601 aaaaacattt attttcattg caatgatgta tttaaattat ttctgaatat tttactaaaa
      661 agggaatgtg ggaggtcagt gcatttaaaa cataaagaaa tgaagagcta gttcaaacct
      721 tgggaaaata cactatatct taaactccat gaaagaaggt gaggctgcaa acagctaatg
      781 cacattggca acagcccctg atgcctatgc cttattcatc cctcagaaaa ggattcaagt
      841 agaggcttga tttggaggtt aaagttttgc tatgctgtat tttagaatta attccatacc
      901 acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa
      961 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa
     1021 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     1081 ggtttgtcca aactcatcaa tgtatcttat catgtctggc cagctagcac tcatttaccc
     1141 ggagacaggg agaggctctt ctgcgtgtag tggttgtgca gagcctcgtg catcacggag
     1201 catgagaaga cgttcccctg ctgccacctg ctcttgtcca cggtgagctt gctgtagagg
     1261 aagaaggagc cgtcggagtc cagcacggga ggcgtggtct tgtagttgtt ctccggctgc
     1321 ccattgctct cccactccac ggcgatgtcg ctgggataga agcctttgac caggcaggtc
     1381 aggctgacct ggttcttggt catctcctcc cgggatgggg gcagggtgta cacctgtggt
     1441 tctcggggct gccctttggc tttggagatg gttttctcga tgggggctgg gagggctttg
     1501 ttggagacct tgcacttgta ctccttgcca ttcagccagt cctggtgcag gacggtgagg
     1561 acgctgacca cacggtacgt gctgttgtac tgctcctccc gcggctttgt cttggcatta
     1621 tgcacctcca cgccgtccac gtaccagttg aacttgacct cagggtcttc gtggctcacg
     1681 tccaccacca cgcatgtgac ctcaggggtc cgggagatca tgagggtgtc cttgggtttt
     1741 ggggggaaga ggaagactga cggtcccccc aggagttcag gtgctgggca cggtgggcat
     1801 gtgtgagttt tgtcagatct tcacttgtcg tcatcgtctt tgtagtcaga accgtacttg
     1861 cgttttacct ggccgggaaa aacatctccc tcggatggtt tagagtcagg gcccatcttg
     1921 ctagacagtg tataggtttc ggactcgctc ttggacttta cgatctcccg gaaccacttc
     1981 tggtattgct tctgcacctg ttgggacagc aagcaataca caagaaagat gaaaaaaccc
     2041 tgcaagctgt tgataatagt aaacaggtag gccatcacct gggctgccgg tccgacctgg
     2101 agcaagccca gacaccaggt acagcccagg ataaacagct gggctgtggc tttgaaggcc
     2161 agcatgcggg tattctggat cgtggacact tcagagttca gagagctaag cttacgtttg
     2221 aggatccaga agaccaaaat gaacagcacc aggttagcag agaaaatagc gcaaacgggc
     2281 cccagaaagc tccacataaa tccctggtcc aggtgcagcc agcaccggtc ggctgtgccg
     2341 tacaggtgtg gccatgatgc agcactgatg gcgacagtca cggcaggcac cccatagccc
     2401 acagggaaca taatccactt catgagtctg ttgatagagc tgtagttaac cacagtcagg
     2461 ttcctagcgg tcaggaacag gtgcacgccc tccagaagca tccaggtaaa ggcagccaga
     2521 taaagataat ggagagcacc ggcgataatg gaacacagaa ccttaggttc agtacgatcg
     2581 atcccaacca ggaacaggag gtgggcgagg aacaggcaca gagacagctg gagatgcagg
     2641 ctggtagaag tgttgcgaat tgccttacac aggaggaaag tgagggcagc caggagaagg
     2701 cacaagaggg agactgagag tcctacgtag gtgataacgg tcagcacagg gtcctcttct
     2761 tgagaagtaa gcgcttggaa gtaaaggttc tcgttggtct gcgcatcttt cagcgcttca
     2821 tccacggtct ggcggccgct cgccgcgtta atcaccgcgg tgcgcaccgc ataccaaaac
     2881 gcgctcatct gcggaatgtt cggcataatt tcgcctttct gcgcgttttc catggtcgcc
     2941 gcaatgcgcg gatctttcgc cagttcttct tcatagcttt tcagcgccac cgcgcccagc
     3001 ggtttatctt tgttcaccgc ttccaggcct tcatcggtca gcagatagtt ttccagaaat
     3061 tctttcgcca gttctttgtt cgggctcgcc gcgttaatgc ccgcgctcag cacgcccaca
     3121 aacggtttgc tcggctggcc tttaaaggtc ggcagcacgg tcacgccata gttcactttg
     3181 ctggtatcaa tgttgctcca cgcccacggg ccgttaatgg tcatcgcggt ttcgcctttg
     3241 ttaaacgccg cttccgcaat gctataatcg gtatccgcgt tcatatgttt gtttttaatc
     3301 agatccacca gaaaggtcag gcccgctttc gcgcccgcgt tatccacgcc cacatcttta
     3361 atatcatatt tgccgttttc atatttaaac gcatagccgc catccgccgc aatcagcggc
     3421 caggtaaaat acggttcctg caggttaaac atcagcgcgc ttttgccttt cgctttcagt
     3481 tctttatcca gcgccggaat ttcttcccag gttttcggcg ggttcggcag cagatctttg
     3541 ttataaatca ggctcagcgc ttccaccgca atcggatacg caatcagttt gccgttatag
     3601 cgcaccgcat cccaggtaaa cggatacagt ttatcctgaa acgctttatc cggggtaatt
     3661 tccgccagca ggccgctctg cgcatagccg ccaaagcgat catgcgccca aaaaataata
     3721 tccgggccat cgccggtcgc cgccacctgc ggaaattttt cttccagttt atccggatgt
     3781 tccacggtca ctttaatgcc ggtatctttt tcaaattttt tgcccacttc cgccaggccg
     3841 ttatagcctt tatcgccgtt aatccaaatc accagtttgc cttcttcaat tttggatcca
     3901 gcgtaatctg gaacatcgta tgggtacgaa ttcgtgacaa gtgcaagact tagtgcaatg
     3961 caagacagga gttgcatcct gtacatggtg gccctccttc gccggtgatc tcaggtaggc
     4021 gccggtcaca gcttggatct gtaacggcgc agaacagaaa acgaaacaaa gacgtagagt
     4081 tgagcaagca gggtcaggca aagcgtggag agccggctga gtctaggtag gctccaaggg
     4141 agcgccggac aaaggcccgg tctcgacctg agctttaaac ttacctagac ggcggacgca
     4201 gttcaggagg caccacaggc gggaggcggc agaacgcgac tcaaccggcg tggatggcgg
     4261 cctcaggtag ggcggcgggc gcgtgaagga gagatgcgag cccctcgaag cttcagctgt
     4321 gttctggcgg caaacccgtt gcgaaaaaga acgttcacgg cgactactgc acttatatac
     4381 ggttctcccc caccctcggg aaaaaggcgg agccagtaca cgacatcact ttcccagttt
     4441 accccgcgcc accttctcta ggcacccgtt caattgccga cccctccccc caacttctcg
     4501 gggactgtgg gcgatgtgcg ctctgcccac tgacgggcac cggagcgatc gcagatcctt
     4561 cgatagagaa atgttctggc acctgcactt gcactgggga cagcctattt tgctagtttg
     4621 ttttgtttcg ttttgttttg atggagagcg tatgttagtt acgattcaca caaaaaacca
     4681 acacacagat gtaatgaaaa taaagatatt ttattgcggc cgctgattta aatgttaatt
     4741 aactagccat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     4801 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     4861 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     4921 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     4981 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     5041 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     5101 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     5161 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     5221 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     5281 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     5341 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     5401 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     5461 ttgctcacat gttcttaatt aacctgcagg cgttacataa cttacggtaa atggcccgcc
     5521 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
     5581 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
     5641 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
     5701 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
     5761 gtacatctac gtattagtca tcgctattac catgatgatg cggttttggc agtacatcaa
     5821 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
     5881 tgggagtttg ttttgactag tcagggcccc aaccccccca agcccccatt tcacaacacg
     5941 ctggcgctac aggcgcgtga cttccccttg ctttggggcg gggggctgag actcctatgt
     6001 gctccggatt ggtcaggcac ggccttcggc cccgcctcct gccaccgcag attggccgct
     6061 aggcctcccc gagcgccctg cctccgaggg ccggcgcacc ataaaagaag ccgccctagc
     6121 cacgtcccct cgcagttcgg cggtcccgcg ggtctgtctc aagcttctgc cttctccctc
     6181 ctgtgagttt ggtaagtcac tgactgtcta tgcctgggaa agggtgggca ggagatgggg
     6241 cagtgcagga aaagtggcac tatgaaccct gcagccctag tttga
//
