LOCUS       sequence-430016-        6486 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 6486)
  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 6486)
  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-ADGRG6 CTFdeltaTA-FLAG
FEATURES             Location/Qualifiers
     source          1..6486
                     /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             2913..2936
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    3651..3772
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3662..3681)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3716..3735
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     polyA_signal    3876..4270
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     CDS             complement(4284..4658)
                     /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        5251..5554
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(5634..6222)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5714..5733)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    6330..6378
                     /note="synthetic polyadenylation signal"
     misc_feature    6389..6480
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     6429..6448
                     /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 ccaagacctt ccaagaagtg
     1801 cctcacagtt agatgcaaga aacactaaag tcctcacttt catcagctat attgggtgtg
     1861 gaatatctgc tattttttca gcagcaactc tcctgacata tgttgctttt gagaaattgc
     1921 gaagggatta tccctccaaa atcttgatga acctgagcac agccctgctg ttcctgaatc
     1981 tcctcttcct cctagatggc tggatcacct ccttcaatgt ggatggactt tgcattgctg
     2041 ttgcagtcct gttgcatttc ttccttctgg caacctttac ctggatgggg ctagaagcaa
     2101 ttcacatgta cattgctcta gttaaagtat ttaacactta cattcgccga tacattctaa
     2161 aattctgcat cattggctgg ggtttgcctg ccttagtggt gtcagttgtt ctagcgagca
     2221 gaaacaacaa tgaagtctat ggaaaagaaa gttatgggaa agaaaaaggt gatgaattct
     2281 gttggattca agatccagtc atattttatg tgacctgtgc tgggtatttt ggagtcatgt
     2341 tttttctgaa cattgccatg ttcattgtgg taatggtgca gatctgtggg aggaatggca
     2401 agagaagcaa ccggaccctg agagaagaag tgttaaggaa cctgcgcagt gtggttagct
     2461 tgacctttct gttgggcatg acatggggtt ttgcattctt tgcctgggga cccttaaata
     2521 tccccttcat gtacctcttc tccatcttca attcattaca aggcttattt atattcatct
     2581 tccactgtgc tatgaaggag aatgttcaga aacagtggcg gcagcatctc tgctgtggta
     2641 gatttcggtt agcagataac tcagattgga gtaagacagc taccaatatc atcaagaaaa
     2701 gttctgataa tctaggaaaa tctttgtctt caagctccat tggttccaac tcaacctatc
     2761 ttacatccaa atctaaatcc agctctacca cctatttcaa aaggaatagt cacacagata
     2821 atgtctccta tgagcattcc ttcaacaaaa gtggatcact cagacagtgc ttccatggac
     2881 aagtccttgt caaaactggc ccatgcggtt ctgactacaa agacgatgac gacaagtgaa
     2941 gatctgacaa aactcacaca tgcccaccgt gcccagcacc tgaactcctg gggggaccgt
     3001 cagtcttcct cttcccccca aaacccaagg acaccctcat gatctcccgg acccctgagg
     3061 tcacatgcgt ggtggtggac gtgagccacg aagaccctga ggtcaagttc aactggtacg
     3121 tggacggcgt ggaggtgcat aatgccaaga caaagccgcg ggaggagcag tacaacagca
     3181 cgtaccgtgt ggtcagcgtc ctcaccgtcc tgcaccagga ctggctgaat ggcaaggagt
     3241 acaagtgcaa ggtctccaac aaagccctcc cagcccccat cgagaaaacc atctccaaag
     3301 ccaaagggca gccccgagaa ccacaggtgt acaccctgcc cccatcccgg gaggagatga
     3361 ccaagaacca ggtcagcctg acctgcctgg tcaaaggctt ctatcccagc gacatcgccg
     3421 tggagtggga gagcaatggg cagccggaga acaactacaa gaccacgcct cccgtgctgg
     3481 actccgacgg ctccttcttc ctctacagca agctcaccgt ggacaagagc aggtggcagc
     3541 aggggaacgt cttctcatgc tccgtgatgc acgaggctct gcacaaccac tacacgcaga
     3601 agagcctctc cctgtctccg ggtaaatgag tgctagctgg ccagacatga taagatacat
     3661 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     3721 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
     3781 caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa
     3841 gtaaaacctc tacaaatgtg gtatggaatt aattctaaaa tacagcatag caaaacttta
     3901 acctccaaat caagcctcta cttgaatcct tttctgaggg atgaataagg cataggcatc
     3961 aggggctgtt gccaatgtgc attagctgtt tgcagcctca ccttctttca tggagtttaa
     4021 gatatagtgt attttcccaa ggtttgaact agctcttcat ttctttatgt tttaaatgca
     4081 ctgacctccc acattccctt tttagtaaaa tattcagaaa taatttaaat acatcattgc
     4141 aatgaaaata aatgtttttt attaggcaga atccagatgc tcaaggccct tcataatatc
     4201 ccccagttta gtagttggac ttagggaaca aaggaacctt taatagaaat tggacagcaa
     4261 gaaagcgagc ttctagctta tcctcagtcc tgctcctctg ccacaaagtg cacgcagttg
     4321 ccggccgggt cgcgcagggc gaactcccgc ccccacggct gctcgccgat ctcggtcatg
     4381 gccggcccgg aggcgtcccg gaagttcgtg gacacgacct ccgaccactc ggcgtacagc
     4441 tcgtccaggc cgcgcaccca cacccaggcc agggtgttgt ccggcaccac ctggtcctgg
     4501 accgcgctga tgaacagggt cacgtcgtcc cggaccacac cggcgaagtc gtcctccacg
     4561 aagtcccggg agaacccgag ccggtcggtc cagaactcga ccgctccggc gacgtcgcgc
     4621 gcggtgagca ccggaacggc actggtcaac ttggccatga tggctcctcc tgtcaggaga
     4681 ggaaagagaa gaaggttagt acaattgcta tagtgagttg tattatacta tgcagatata
     4741 ctatgccaat gattaattgt caaactaggg ctgcagggtt catagtgcca cttttcctgc
     4801 actgccccat ctcctgccca ccctttccca ggcatagaca gtcagtgact taccaaactc
     4861 acaggaggga gaaggcagaa gcttgagaca gacccgcggg accgccgaac tgcgagggga
     4921 cgtggctagg gcggcttctt ttatggtgcg ccggccctcg gaggcagggc gctcggggag
     4981 gcctagcggc caatctgcgg tggcaggagg cggggccgaa ggccgtgcct gaccaatccg
     5041 gagcacatag gagtctcagc cccccgcccc aaagcaaggg gaagtcacgc gcctgtagcg
     5101 ccagcgtgtt gtgaaatggg ggcttggggg ggttggggcc ctgactagtc aaaacaaact
     5161 cccattgacg tcaatggggt ggagacttgg aaatccccgt gagtcaaacc gctatccacg
     5221 cccattgatg tactgccaaa accgcatcat catggtaata gcgatgacta atacgtagat
     5281 gtactgccaa gtaggaaagt cccataaggt catgtactgg gcataatgcc aggcgggcca
     5341 tttaccgtca ttgacgtcaa tagggggcgt acttggcata tgatacactt gatgtactgc
     5401 caagtgggca gtttaccgta aatactccac ccattgacgt caatggaaag tccctattgg
     5461 cgttactatg ggaacatacg tcattattga cgtcaatggg cgggggtcgt tgggcggtca
     5521 gccaggcggg ccatttaccg taagttatgt aacgcctgca ggttaattaa gaacatgtga
     5581 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     5641 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     5701 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     5761 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     5821 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     5881 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     5941 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     6001 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     6061 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     6121 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     6181 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     6241 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatggct
     6301 agttaattaa catttaaatc agcggccgca ataaaatatc tttattttca ttacatctgt
     6361 gtgttggttt tttgtgtgaa tcgtaactaa catacgctct ccatcaaaac aaaacgaaac
     6421 aaaacaaact agcaaaatag gctgtcccca gtgcaagtgc aggtgccaga acatttctct
     6481 atcgaa
//
