LOCUS       sequence-430001-        6324 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 6324)
  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 6324)
  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-ADGRF3 CTFdeltaTA-FLAG
FEATURES             Location/Qualifiers
     source          1..6324
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        31..242
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             255..523
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     sig_peptide     573..632
                     /codon_start=1
                     /product="human interleukin-2 signal sequence"
                     /label=IL-2 signal sequence
                     /translation="MYRMQLLSCIALSLALVTNS"
     CDS             633..659
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1767..1787
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             2751..2774
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    3489..3610
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3500..3519)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3554..3573
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     polyA_signal    3714..4108
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     CDS             complement(4122..4496)
                     /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        5089..5392
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(5472..6060)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5552..5571)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    6168..6216
                     /note="synthetic polyadenylation signal"
     misc_feature    6227..6318
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     6267..6286
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggatctgcga tcgctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg
       61 agaagttggg gggaggggtc ggcaattgaa cgggtgccta gagaaggtgg cgcggggtaa
      121 actgggaaag tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt
      181 atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac
      241 agctgaagct tcgaggggct cgcatctctc cttcacgcgc ccgccgccct acctgaggcc
      301 gccatccacg ccggttgagt cgcgttctgc cgcctcccgc ctgtggtgcc tcctgaactg
      361 cgtccgccgt ctaggtaagt ttaaagctca ggtcgagacc gggcctttgt ccggcgctcc
      421 cttggagcct acctagactc agccggctct ccacgctttg cctgaccctg cttgctcaac
      481 tctacgtctt tgtttcgttt tctgttctgc gccgttacag atccaagctg tgaccggcgc
      541 ctacctgaga tcaccggcga aggagggcca ccatgtacag gatgcaactc ctgtcttgca
      601 ttgcactaag tcttgcactt gtcacgaatt cgtacccata cgatgttcca gattacgctg
      661 gatccaaaat tgaagaaggc aaactggtga tttggattaa cggcgataaa ggctataacg
      721 gcctggcgga agtgggcaaa aaatttgaaa aagataccgg cattaaagtg accgtggaac
      781 atccggataa actggaagaa aaatttccgc aggtggcggc gaccggcgat ggcccggata
      841 ttattttttg ggcgcatgat cgctttggcg gctatgcgca gagcggcctg ctggcggaaa
      901 ttaccccgga taaagcgttt caggataaac tgtatccgtt tacctgggat gcggtgcgct
      961 ataacggcaa actgattgcg tatccgattg cggtggaagc gctgagcctg atttataaca
     1021 aagatctgct gccgaacccg ccgaaaacct gggaagaaat tccggcgctg gataaagaac
     1081 tgaaagcgaa aggcaaaagc gcgctgatgt ttaacctgca ggaaccgtat tttacctggc
     1141 cgctgattgc ggcggatggc ggctatgcgt ttaaatatga aaacggcaaa tatgatatta
     1201 aagatgtggg cgtggataac gcgggcgcga aagcgggcct gacctttctg gtggatctga
     1261 ttaaaaacaa acatatgaac gcggataccg attatagcat tgcggaagcg gcgtttaaca
     1321 aaggcgaaac cgcgatgacc attaacggcc cgtgggcgtg gagcaacatt gataccagca
     1381 aagtgaacta tggcgtgacc gtgctgccga cctttaaagg ccagccgagc aaaccgtttg
     1441 tgggcgtgct gagcgcgggc attaacgcgg cgagcccgaa caaagaactg gcgaaagaat
     1501 ttctggaaaa ctatctgctg accgatgaag gcctggaagc ggtgaacaaa gataaaccgc
     1561 tgggcgcggt ggcgctgaaa agctatgaag aagaactggc gaaagatccg cgcattgcgg
     1621 cgaccatgga aaacgcgcag aaaggcgaaa ttatgccgaa cattccgcag atgagcgcgt
     1681 tttggtatgc ggtgcgcacc gcggtgatta acgcggcgag cggccgccag accgtggatg
     1741 aagcgctgaa agatgcgcag accaacgaga acctttactt ccaaagcccc cacaccgtgc
     1801 cggaggaacc cgcgcttgcg ctgctcactc aggttggact gggcgccagc atcttggcac
     1861 ttctggtctg cctgggagtt tactggctgg tgtggcgggt tgtcgtgcgc aacaaaattt
     1921 cttatttccg ccacgccgcg ctgttgaaca tggtgttttg cctcctggct gccgacacct
     1981 gtttcctggg cgcccccttc ctgagccccg ggccccgctc tcctctgtgc ctggccgctg
     2041 cctttctgtg ccactttctg tacctggcaa catttttctg gatgcttgcc caggctctgg
     2101 tgctggctca ccagcttctg ttcgtgtttc accagctcgc aaaacacaga gtgctgcccc
     2161 tgatggtgct cctgggatac ctgtgccccc ttggcttggc tggcgttaca ctgggcctgt
     2221 acctccctca gggccagtat ctgcgcgagg gcgagtgctg gctggacggt aagggcggag
     2281 cactgtacac ctttgtgggc cccgttctgg cgattatcgg agtcaacggg ctggtcttgg
     2341 ccatggcaat gctcaaattg ctgaggcctt ctctttccga gggtccgcca gcagaaaagc
     2401 gccaggccct cctgggcgtc atcaaggccc tccttatcct caccccaatc ttcgggttga
     2461 cctggggcct gggtcttgcg accctgctcg aagaggttag taccgtgccc cactatatct
     2521 tcaccatcct gaatacactg cagggcgtct ttatcctgct tttcggctgc ctgatggacc
     2581 ggaaaatcca ggaagcgttg cgtaagcgct tttgccgcgc ccaggcaccg agctctacta
     2641 tcagtctcgt gagttgttgc ctgcagatcc tgagctgtgc aagcaaatct atgtctgagg
     2701 gtatcccatg gccatcctct gaagatatgg gtaccgcccg cagtggttct gactacaaag
     2761 acgatgacga caagtgaaga tctgacaaaa ctcacacatg cccaccgtgc ccagcacctg
     2821 aactcctggg gggaccgtca gtcttcctct tccccccaaa acccaaggac accctcatga
     2881 tctcccggac ccctgaggtc acatgcgtgg tggtggacgt gagccacgaa gaccctgagg
     2941 tcaagttcaa ctggtacgtg gacggcgtgg aggtgcataa tgccaagaca aagccgcggg
     3001 aggagcagta caacagcacg taccgtgtgg tcagcgtcct caccgtcctg caccaggact
     3061 ggctgaatgg caaggagtac aagtgcaagg tctccaacaa agccctccca gcccccatcg
     3121 agaaaaccat ctccaaagcc aaagggcagc cccgagaacc acaggtgtac accctgcccc
     3181 catcccggga ggagatgacc aagaaccagg tcagcctgac ctgcctggtc aaaggcttct
     3241 atcccagcga catcgccgtg gagtgggaga gcaatgggca gccggagaac aactacaaga
     3301 ccacgcctcc cgtgctggac tccgacggct ccttcttcct ctacagcaag ctcaccgtgg
     3361 acaagagcag gtggcagcag gggaacgtct tctcatgctc cgtgatgcac gaggctctgc
     3421 acaaccacta cacgcagaag agcctctccc tgtctccggg taaatgagtg ctagctggcc
     3481 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     3541 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     3601 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg
     3661 ggaggttttt taaagcaagt aaaacctcta caaatgtggt atggaattaa ttctaaaata
     3721 cagcatagca aaactttaac ctccaaatca agcctctact tgaatccttt tctgagggat
     3781 gaataaggca taggcatcag gggctgttgc caatgtgcat tagctgtttg cagcctcacc
     3841 ttctttcatg gagtttaaga tatagtgtat tttcccaagg tttgaactag ctcttcattt
     3901 ctttatgttt taaatgcact gacctcccac attccctttt tagtaaaata ttcagaaata
     3961 atttaaatac atcattgcaa tgaaaataaa tgttttttat taggcagaat ccagatgctc
     4021 aaggcccttc ataatatccc ccagtttagt agttggactt agggaacaaa ggaaccttta
     4081 atagaaattg gacagcaaga aagcgagctt ctagcttatc ctcagtcctg ctcctctgcc
     4141 acaaagtgca cgcagttgcc ggccgggtcg cgcagggcga actcccgccc ccacggctgc
     4201 tcgccgatct cggtcatggc cggcccggag gcgtcccgga agttcgtgga cacgacctcc
     4261 gaccactcgg cgtacagctc gtccaggccg cgcacccaca cccaggccag ggtgttgtcc
     4321 ggcaccacct ggtcctggac cgcgctgatg aacagggtca cgtcgtcccg gaccacaccg
     4381 gcgaagtcgt cctccacgaa gtcccgggag aacccgagcc ggtcggtcca gaactcgacc
     4441 gctccggcga cgtcgcgcgc ggtgagcacc ggaacggcac tggtcaactt ggccatgatg
     4501 gctcctcctg tcaggagagg aaagagaaga aggttagtac aattgctata gtgagttgta
     4561 ttatactatg cagatatact atgccaatga ttaattgtca aactagggct gcagggttca
     4621 tagtgccact tttcctgcac tgccccatct cctgcccacc ctttcccagg catagacagt
     4681 cagtgactta ccaaactcac aggagggaga aggcagaagc ttgagacaga cccgcgggac
     4741 cgccgaactg cgaggggacg tggctagggc ggcttctttt atggtgcgcc ggccctcgga
     4801 ggcagggcgc tcggggaggc ctagcggcca atctgcggtg gcaggaggcg gggccgaagg
     4861 ccgtgcctga ccaatccgga gcacatagga gtctcagccc cccgccccaa agcaagggga
     4921 agtcacgcgc ctgtagcgcc agcgtgttgt gaaatggggg cttggggggg ttggggccct
     4981 gactagtcaa aacaaactcc cattgacgtc aatggggtgg agacttggaa atccccgtga
     5041 gtcaaaccgc tatccacgcc cattgatgta ctgccaaaac cgcatcatca tggtaatagc
     5101 gatgactaat acgtagatgt actgccaagt aggaaagtcc cataaggtca tgtactgggc
     5161 ataatgccag gcgggccatt taccgtcatt gacgtcaata gggggcgtac ttggcatatg
     5221 atacacttga tgtactgcca agtgggcagt ttaccgtaaa tactccaccc attgacgtca
     5281 atggaaagtc cctattggcg ttactatggg aacatacgtc attattgacg tcaatgggcg
     5341 ggggtcgttg ggcggtcagc caggcgggcc atttaccgta agttatgtaa cgcctgcagg
     5401 ttaattaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     5461 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     5521 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     5581 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     5641 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     5701 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     5761 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     5821 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     5881 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     5941 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     6001 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     6061 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     6121 gggattttgg tcatggctag ttaattaaca tttaaatcag cggccgcaat aaaatatctt
     6181 tattttcatt acatctgtgt gttggttttt tgtgtgaatc gtaactaaca tacgctctcc
     6241 atcaaaacaa aacgaaacaa aacaaactag caaaataggc tgtccccagt gcaagtgcag
     6301 gtgccagaac atttctctat cgaa
//
