LOCUS       sequence-430296-        7269 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 7269)
  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 7269)
  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-ADGRL2 CTF-FLAG
FEATURES             Location/Qualifiers
     source          1..7269
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(102..690)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(182..201)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    798..846
                     /note="synthetic polyadenylation signal"
     misc_feature    857..948
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     897..916
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        985..1196
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             1209..1477
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     sig_peptide     1527..1586
                     /codon_start=1
                     /product="human interleukin-2 signal sequence"
                     /label=IL-2 signal sequence
                     /translation="MYRMQLLSCIALSLALVTNS"
     CDS             1587..1613
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4650..4673
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    5388..5509
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5399..5418)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5453..5472
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     polyA_signal    5613..6007
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     CDS             complement(6021..6395)
                     /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        join(6988..7269,1..22)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 atttaccgta agttatgtaa cgcctgcagg ttaattaaga acatgtgagc aaaaggccag
       61 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
      121 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
      181 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
      241 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
      301 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
      361 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
      421 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
      481 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
      541 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
      601 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
      661 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
      721 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatggctag ttaattaaca
      781 tttaaatcag cggccgcaat aaaatatctt tattttcatt acatctgtgt gttggttttt
      841 tgtgtgaatc gtaactaaca tacgctctcc atcaaaacaa aacgaaacaa aacaaactag
      901 caaaataggc tgtccccagt gcaagtgcag gtgccagaac atttctctat cgaaggatct
      961 gcgatcgctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc cccgagaagt
     1021 tggggggagg ggtcggcaat tgaacgggtg cctagagaag gtggcgcggg gtaaactggg
     1081 aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa ccgtatataa
     1141 gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga acacagctga
     1201 agcttcgagg ggctcgcatc tctccttcac gcgcccgccg ccctacctga ggccgccatc
     1261 cacgccggtt gagtcgcgtt ctgccgcctc ccgcctgtgg tgcctcctga actgcgtccg
     1321 ccgtctaggt aagtttaaag ctcaggtcga gaccgggcct ttgtccggcg ctcccttgga
     1381 gcctacctag actcagccgg ctctccacgc tttgcctgac cctgcttgct caactctacg
     1441 tctttgtttc gttttctgtt ctgcgccgtt acagatccaa gctgtgaccg gcgcctacct
     1501 gagatcaccg gcgaaggagg gccaccatgt acaggatgca actcctgtct tgcattgcac
     1561 taagtcttgc acttgtcacg aattcgtacc catacgatgt tccagattac gctggatcca
     1621 aaattgaaga aggcaaactg gtgatttgga ttaacggcga taaaggctat aacggcctgg
     1681 cggaagtggg caaaaaattt gaaaaagata ccggcattaa agtgaccgtg gaacatccgg
     1741 ataaactgga agaaaaattt ccgcaggtgg cggcgaccgg cgatggcccg gatattattt
     1801 tttgggcgca tgatcgcttt ggcggctatg cgcagagcgg cctgctggcg gaaattaccc
     1861 cggataaagc gtttcaggat aaactgtatc cgtttacctg ggatgcggtg cgctataacg
     1921 gcaaactgat tgcgtatccg attgcggtgg aagcgctgag cctgatttat aacaaagatc
     1981 tgctgccgaa cccgccgaaa acctgggaag aaattccggc gctggataaa gaactgaaag
     2041 cgaaaggcaa aagcgcgctg atgtttaacc tgcaggaacc gtattttacc tggccgctga
     2101 ttgcggcgga tggcggctat gcgtttaaat atgaaaacgg caaatatgat attaaagatg
     2161 tgggcgtgga taacgcgggc gcgaaagcgg gcctgacctt tctggtggat ctgattaaaa
     2221 acaaacatat gaacgcggat accgattata gcattgcgga agcggcgttt aacaaaggcg
     2281 aaaccgcgat gaccattaac ggcccgtggg cgtggagcaa cattgatacc agcaaagtga
     2341 actatggcgt gaccgtgctg ccgaccttta aaggccagcc gagcaaaccg tttgtgggcg
     2401 tgctgagcgc gggcattaac gcggcgagcc cgaacaaaga actggcgaaa gaatttctgg
     2461 aaaactatct gctgaccgat gaaggcctgg aagcggtgaa caaagataaa ccgctgggcg
     2521 cggtggcgct gaaaagctat gaagaagaac tggcgaaaga tccgcgcatt gcggcgacca
     2581 tggaaaacgc gcagaaaggc gaaattatgc cgaacattcc gcagatgagc gcgttttggt
     2641 atgcggtgcg caccgcggtg attaacgcgg cgagcggccg ccagaccgtg gatgaagcgc
     2701 tgaaagatgc gcagaccaac gagaaccttt acttccaaac caacttcgct attctgatgg
     2761 cccaccggga gattgcttac aaggatggtg tgcacgagct tttgctcacc gtcatcacct
     2821 gggtgggtat tgtgatcagt ttggtttgcc tcgccatctg catttttact ttttgttttt
     2881 tcaggggtct tcagtctgac cggaatacta tccacaagaa cttgtgcatc aacctgttca
     2941 ttgccgagtt catcttcttg atcggcatcg acaaaacgaa atacgccatc gcctgtccga
     3001 tctttgctgg tctgctccac tttttctttc tggcagcctt tgcttggatg tgcctggaag
     3061 gcgttcagct gtacctgatg ctggtggagg tgtttgaatc cgagtattct cgcaaaaagt
     3121 attactatgt tgctggttac ctgttccctg ccaccgtcgt gggcgtatct gcagctatcg
     3181 actataaaag ttacggtacg gagaaggcct gctggcttca cgtcgacaac tatttcatct
     3241 ggtcattcat cggccccgtg acgttcatta tccttctgaa cattatcttc cttgtcatca
     3301 ctctctgtaa aatggtcaag cacagcaaca cactgaagcc tgactcatct cgcctggaga
     3361 acatcaagag ttgggttctg ggcgcattcg ccctgttgtg tcttctgggg ctgacctggt
     3421 ccttcggctt gctcttcatt aacgaagaga ccatcgttat ggcctacctt ttcaccatct
     3481 tcaacgcttt ccagggcgtc tttatcttta tcttccactg tgcattgcag aaaaaggttc
     3541 gtaaagagta cggtaagtgt tttaggcatt cctattgctg tggaggcctg cccacagagt
     3601 ctccacacag ctcagtaaaa gcttctacca ctagaacctc cgcccgctac tctagcggta
     3661 cccagtcaag aatcaggcgc atgtggaacg ataccgtgag aaaacagtcc gagtctagct
     3721 tcatctccgg agacattaac tccacgtcaa ccctgaacca gggcatgacc ggcaactatc
     3781 tgcttacaaa ccctcttctg cgtccccacg ggactaataa cccatacaat acgcttctgg
     3841 ccgagaccgt ggtatgcaac gcacccagtg cccctgtctt caattctccg ggtcacagcc
     3901 tgaataacgc aagagacacg agtgctatgg acaccctgcc actgaacggc aactttaata
     3961 actcatactc actgcacaaa ggtgattaca acgactccgt gcaggtcgtt gattgcggcc
     4021 tgtctctgaa cgacacagcc ttcgagaaga tgattatctc tgagctggtc cacaataacc
     4081 tgagaggcag ctccaagacc cataacctgg agctgactct gcccgtgaaa cccgttattg
     4141 gtggctccag ctcagaggat gacgctatcg tggccgacgc ctcatccctg atgcattccg
     4201 acaaccctgg gctggagctc catcacaagg agctcgaagc tcccctgatc ccgcagagaa
     4261 cccattctct cctgtaccag ccccagaaaa aggtaaaaag tgagggtacg gatagctatg
     4321 tgtcacagct gaccgccgag gccgaggacc atctgcagag ccccaatcgc gacagcctct
     4381 acacatccat gcccaacctg cgtgactcac cctatcccga atcttccccc gatatggaag
     4441 aggatctgag cccgtcccgt cgctctgaga acgaagatat ctactataag agcatgccga
     4501 atctgggagc cggtcatcag cttcaaatgt gctatcagat ctctcgggga aattctgacg
     4561 gctacattat ccctatcaac aaagaaggat gtatcccaga aggcgacgtg cgcgaaggac
     4621 aaatgcagct ggtcacctct ctgggttctg actacaaaga cgatgacgac aagtgaagat
     4681 ctgacaaaac tcacacatgc ccaccgtgcc cagcacctga actcctgggg ggaccgtcag
     4741 tcttcctctt ccccccaaaa cccaaggaca ccctcatgat ctcccggacc cctgaggtca
     4801 catgcgtggt ggtggacgtg agccacgaag accctgaggt caagttcaac tggtacgtgg
     4861 acggcgtgga ggtgcataat gccaagacaa agccgcggga ggagcagtac aacagcacgt
     4921 accgtgtggt cagcgtcctc accgtcctgc accaggactg gctgaatggc aaggagtaca
     4981 agtgcaaggt ctccaacaaa gccctcccag cccccatcga gaaaaccatc tccaaagcca
     5041 aagggcagcc ccgagaacca caggtgtaca ccctgccccc atcccgggag gagatgacca
     5101 agaaccaggt cagcctgacc tgcctggtca aaggcttcta tcccagcgac atcgccgtgg
     5161 agtgggagag caatgggcag ccggagaaca actacaagac cacgcctccc gtgctggact
     5221 ccgacggctc cttcttcctc tacagcaagc tcaccgtgga caagagcagg tggcagcagg
     5281 ggaacgtctt ctcatgctcc gtgatgcacg aggctctgca caaccactac acgcagaaga
     5341 gcctctccct gtctccgggt aaatgagtgc tagctggcca gacatgataa gatacattga
     5401 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     5461 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
     5521 ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta
     5581 aaacctctac aaatgtggta tggaattaat tctaaaatac agcatagcaa aactttaacc
     5641 tccaaatcaa gcctctactt gaatcctttt ctgagggatg aataaggcat aggcatcagg
     5701 ggctgttgcc aatgtgcatt agctgtttgc agcctcacct tctttcatgg agtttaagat
     5761 atagtgtatt ttcccaaggt ttgaactagc tcttcatttc tttatgtttt aaatgcactg
     5821 acctcccaca ttcccttttt agtaaaatat tcagaaataa tttaaataca tcattgcaat
     5881 gaaaataaat gttttttatt aggcagaatc cagatgctca aggcccttca taatatcccc
     5941 cagtttagta gttggactta gggaacaaag gaacctttaa tagaaattgg acagcaagaa
     6001 agcgagcttc tagcttatcc tcagtcctgc tcctctgcca caaagtgcac gcagttgccg
     6061 gccgggtcgc gcagggcgaa ctcccgcccc cacggctgct cgccgatctc ggtcatggcc
     6121 ggcccggagg cgtcccggaa gttcgtggac acgacctccg accactcggc gtacagctcg
     6181 tccaggccgc gcacccacac ccaggccagg gtgttgtccg gcaccacctg gtcctggacc
     6241 gcgctgatga acagggtcac gtcgtcccgg accacaccgg cgaagtcgtc ctccacgaag
     6301 tcccgggaga acccgagccg gtcggtccag aactcgaccg ctccggcgac gtcgcgcgcg
     6361 gtgagcaccg gaacggcact ggtcaacttg gccatgatgg ctcctcctgt caggagagga
     6421 aagagaagaa ggttagtaca attgctatag tgagttgtat tatactatgc agatatacta
     6481 tgccaatgat taattgtcaa actagggctg cagggttcat agtgccactt ttcctgcact
     6541 gccccatctc ctgcccaccc tttcccaggc atagacagtc agtgacttac caaactcaca
     6601 ggagggagaa ggcagaagct tgagacagac ccgcgggacc gccgaactgc gaggggacgt
     6661 ggctagggcg gcttctttta tggtgcgccg gccctcggag gcagggcgct cggggaggcc
     6721 tagcggccaa tctgcggtgg caggaggcgg ggccgaaggc cgtgcctgac caatccggag
     6781 cacataggag tctcagcccc ccgccccaaa gcaaggggaa gtcacgcgcc tgtagcgcca
     6841 gcgtgttgtg aaatgggggc ttgggggggt tggggccctg actagtcaaa acaaactccc
     6901 attgacgtca atggggtgga gacttggaaa tccccgtgag tcaaaccgct atccacgccc
     6961 attgatgtac tgccaaaacc gcatcatcat ggtaatagcg atgactaata cgtagatgta
     7021 ctgccaagta ggaaagtccc ataaggtcat gtactgggca taatgccagg cgggccattt
     7081 accgtcattg acgtcaatag ggggcgtact tggcatatga tacacttgat gtactgccaa
     7141 gtgggcagtt taccgtaaat actccaccca ttgacgtcaa tggaaagtcc ctattggcgt
     7201 tactatggga acatacgtca ttattgacgt caatgggcgg gggtcgttgg gcggtcagcc
     7261 aggcgggcc
//
