LOCUS       sequence-430279-        6270 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 6270)
  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 6270)
  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-ADGRE1 CTF-FLAG
FEATURES             Location/Qualifiers
     source          1..6270
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    10..131
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(21..40)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     75..94
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     polyA_signal    235..629
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     CDS             complement(643..1017)
                     /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        1610..1913
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(1993..2581)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2073..2092)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    2689..2737
                     /note="synthetic polyadenylation signal"
     misc_feature    2748..2839
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     2788..2807
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        2876..3087
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             3100..3368
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     sig_peptide     3418..3477
                     /codon_start=1
                     /product="human interleukin-2 signal sequence"
                     /label=IL-2 signal sequence
                     /translation="MYRMQLLSCIALSLALVTNS"
     CDS             3478..3504
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4612..4632
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQA"
     CDS             5542..5565
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
ORIGIN
        1 cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa
       61 aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca
      121 ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt
      181 gggaggtttt ttaaagcaag taaaacctct acaaatgtgg tatggaatta attctaaaat
      241 acagcatagc aaaactttaa cctccaaatc aagcctctac ttgaatcctt ttctgaggga
      301 tgaataaggc ataggcatca ggggctgttg ccaatgtgca ttagctgttt gcagcctcac
      361 cttctttcat ggagtttaag atatagtgta ttttcccaag gtttgaacta gctcttcatt
      421 tctttatgtt ttaaatgcac tgacctccca cattcccttt ttagtaaaat attcagaaat
      481 aatttaaata catcattgca atgaaaataa atgtttttta ttaggcagaa tccagatgct
      541 caaggccctt cataatatcc cccagtttag tagttggact tagggaacaa aggaaccttt
      601 aatagaaatt ggacagcaag aaagcgagct tctagcttat cctcagtcct gctcctctgc
      661 cacaaagtgc acgcagttgc cggccgggtc gcgcagggcg aactcccgcc cccacggctg
      721 ctcgccgatc tcggtcatgg ccggcccgga ggcgtcccgg aagttcgtgg acacgacctc
      781 cgaccactcg gcgtacagct cgtccaggcc gcgcacccac acccaggcca gggtgttgtc
      841 cggcaccacc tggtcctgga ccgcgctgat gaacagggtc acgtcgtccc ggaccacacc
      901 ggcgaagtcg tcctccacga agtcccggga gaacccgagc cggtcggtcc agaactcgac
      961 cgctccggcg acgtcgcgcg cggtgagcac cggaacggca ctggtcaact tggccatgat
     1021 ggctcctcct gtcaggagag gaaagagaag aaggttagta caattgctat agtgagttgt
     1081 attatactat gcagatatac tatgccaatg attaattgtc aaactagggc tgcagggttc
     1141 atagtgccac ttttcctgca ctgccccatc tcctgcccac cctttcccag gcatagacag
     1201 tcagtgactt accaaactca caggagggag aaggcagaag cttgagacag acccgcggga
     1261 ccgccgaact gcgaggggac gtggctaggg cggcttcttt tatggtgcgc cggccctcgg
     1321 aggcagggcg ctcggggagg cctagcggcc aatctgcggt ggcaggaggc ggggccgaag
     1381 gccgtgcctg accaatccgg agcacatagg agtctcagcc ccccgcccca aagcaagggg
     1441 aagtcacgcg cctgtagcgc cagcgtgttg tgaaatgggg gcttgggggg gttggggccc
     1501 tgactagtca aaacaaactc ccattgacgt caatggggtg gagacttgga aatccccgtg
     1561 agtcaaaccg ctatccacgc ccattgatgt actgccaaaa ccgcatcatc atggtaatag
     1621 cgatgactaa tacgtagatg tactgccaag taggaaagtc ccataaggtc atgtactggg
     1681 cataatgcca ggcgggccat ttaccgtcat tgacgtcaat agggggcgta cttggcatat
     1741 gatacacttg atgtactgcc aagtgggcag tttaccgtaa atactccacc cattgacgtc
     1801 aatggaaagt ccctattggc gttactatgg gaacatacgt cattattgac gtcaatgggc
     1861 gggggtcgtt gggcggtcag ccaggcgggc catttaccgt aagttatgta acgcctgcag
     1921 gttaattaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     1981 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     2041 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     2101 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     2161 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     2221 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     2281 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     2341 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     2401 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     2461 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     2521 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     2581 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     2641 agggattttg gtcatggcta gttaattaac atttaaatca gcggccgcaa taaaatatct
     2701 ttattttcat tacatctgtg tgttggtttt ttgtgtgaat cgtaactaac atacgctctc
     2761 catcaaaaca aaacgaaaca aaacaaacta gcaaaatagg ctgtccccag tgcaagtgca
     2821 ggtgccagaa catttctcta tcgaaggatc tgcgatcgct ccggtgcccg tcagtgggca
     2881 gagcgcacat cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgaacgggt
     2941 gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt
     3001 tttcccgagg gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt
     3061 cgcaacgggt ttgccgccag aacacagctg aagcttcgag gggctcgcat ctctccttca
     3121 cgcgcccgcc gccctacctg aggccgccat ccacgccggt tgagtcgcgt tctgccgcct
     3181 cccgcctgtg gtgcctcctg aactgcgtcc gccgtctagg taagtttaaa gctcaggtcg
     3241 agaccgggcc tttgtccggc gctcccttgg agcctaccta gactcagccg gctctccacg
     3301 ctttgcctga ccctgcttgc tcaactctac gtctttgttt cgttttctgt tctgcgccgt
     3361 tacagatcca agctgtgacc ggcgcctacc tgagatcacc ggcgaaggag ggccaccatg
     3421 tacaggatgc aactcctgtc ttgcattgca ctaagtcttg cacttgtcac gaattcgtac
     3481 ccatacgatg ttccagatta cgctggatcc aaaattgaag aaggcaaact ggtgatttgg
     3541 attaacggcg ataaaggcta taacggcctg gcggaagtgg gcaaaaaatt tgaaaaagat
     3601 accggcatta aagtgaccgt ggaacatccg gataaactgg aagaaaaatt tccgcaggtg
     3661 gcggcgaccg gcgatggccc ggatattatt ttttgggcgc atgatcgctt tggcggctat
     3721 gcgcagagcg gcctgctggc ggaaattacc ccggataaag cgtttcagga taaactgtat
     3781 ccgtttacct gggatgcggt gcgctataac ggcaaactga ttgcgtatcc gattgcggtg
     3841 gaagcgctga gcctgattta taacaaagat ctgctgccga acccgccgaa aacctgggaa
     3901 gaaattccgg cgctggataa agaactgaaa gcgaaaggca aaagcgcgct gatgtttaac
     3961 ctgcaggaac cgtattttac ctggccgctg attgcggcgg atggcggcta tgcgtttaaa
     4021 tatgaaaacg gcaaatatga tattaaagat gtgggcgtgg ataacgcggg cgcgaaagcg
     4081 ggcctgacct ttctggtgga tctgattaaa aacaaacata tgaacgcgga taccgattat
     4141 agcattgcgg aagcggcgtt taacaaaggc gaaaccgcga tgaccattaa cggcccgtgg
     4201 gcgtggagca acattgatac cagcaaagtg aactatggcg tgaccgtgct gccgaccttt
     4261 aaaggccagc cgagcaaacc gtttgtgggc gtgctgagcg cgggcattaa cgcggcgagc
     4321 ccgaacaaag aactggcgaa agaatttctg gaaaactatc tgctgaccga tgaaggcctg
     4381 gaagcggtga acaaagataa accgctgggc gcggtggcgc tgaaaagcta tgaagaagaa
     4441 ctggcgaaag atccgcgcat tgcggcgacc atggaaaacg cgcagaaagg cgaaattatg
     4501 ccgaacattc cgcagatgag cgcgttttgg tatgcggtgc gcaccgcggt gattaacgcg
     4561 gcgagcggcc gccagaccgt ggatgaagcg ctgaaagatg cgcagaccaa cgagaacctt
     4621 tacttccaag ctaatctggc ggtcatcatg gcttctggcg aactgaccat ggacttcagc
     4681 ctttacatta tctctcacgt tggcattatc atttccttgg tgtgcctcgt cctggctatc
     4741 gctacattcc tcctgtgtag atctatccgc aaccataaca cttacctcca cctgcacctc
     4801 tgtgtgtgtc tgcttctggc caagaccctg tttctcgccg ggattcacaa gaccgacaac
     4861 aaaatgggtt gtgctattat cgctggcttc ctgcattatc tcttcctggc gtgctttttc
     4921 tggatgctgg tggaggctgt catcctgttc ctgatggtca ggaacctcaa agtagtcaac
     4981 tatttcagct cccggaacat caaaatgttg cacatctgcg cgttcggcta cggcctcccg
     5041 atgctggtcg tagtgatttc cgcttccgtc cagccccagg gctacggcat gcataacaga
     5101 tgctggctga acacggaaac aggttttatc tggtctttcc tgggaccagt ttgcactgtt
     5161 atcgtgatca acagtctctt gctcacctgg actctgtgga tcctccggca aaggttgtcc
     5221 agcgtgaacg ctgaggtttc caccctgaag gacacccgcc tcctgacctt caaggctttc
     5281 gctcagctct ttatcctggg ctgcagctgg gtcctcggca tctttcagat cggccccgtg
     5341 gcaggagtca tggcgtacct gtttaccatt atcaactccc tgcagggcgc cttcatcttc
     5401 cttattcact gcctcctgaa tggacaggtt cgtgaagagt acaaaagatg gatcactggc
     5461 aagaccaagc cgagctctca gagccaaaca tctagaatcc tgctcagcag tatgcctagt
     5521 gctagtaaga cgggcggttc tgactacaaa gacgatgacg acaagtgaag atctgacaaa
     5581 actcacacat gcccaccgtg cccagcacct gaactcctgg ggggaccgtc agtcttcctc
     5641 ttccccccaa aacccaagga caccctcatg atctcccgga cccctgaggt cacatgcgtg
     5701 gtggtggacg tgagccacga agaccctgag gtcaagttca actggtacgt ggacggcgtg
     5761 gaggtgcata atgccaagac aaagccgcgg gaggagcagt acaacagcac gtaccgtgtg
     5821 gtcagcgtcc tcaccgtcct gcaccaggac tggctgaatg gcaaggagta caagtgcaag
     5881 gtctccaaca aagccctccc agcccccatc gagaaaacca tctccaaagc caaagggcag
     5941 ccccgagaac cacaggtgta caccctgccc ccatcccggg aggagatgac caagaaccag
     6001 gtcagcctga cctgcctggt caaaggcttc tatcccagcg acatcgccgt ggagtgggag
     6061 agcaatgggc agccggagaa caactacaag accacgcctc ccgtgctgga ctccgacggc
     6121 tccttcttcc tctacagcaa gctcaccgtg gacaagagca ggtggcagca ggggaacgtc
     6181 ttctcatgct ccgtgatgca cgaggctctg cacaaccact acacgcagaa gagcctctcc
     6241 ctgtctccgg gtaaatgagt gctagctggc
//
