LOCUS       sequence-430291-        6507 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 6507)
  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 6507)
  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 CTF-FLAG
FEATURES             Location/Qualifiers
     source          1..6507
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    129..250
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(140..159)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     194..213
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     polyA_signal    354..748
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     CDS             complement(762..1136)
                     /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        1729..2032
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(2112..2700)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2192..2211)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    2808..2856
                     /note="synthetic polyadenylation signal"
     misc_feature    2867..2958
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     2907..2926
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        2995..3206
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             3219..3487
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     sig_peptide     3537..3596
                     /codon_start=1
                     /product="human interleukin-2 signal sequence"
                     /label=IL-2 signal sequence
                     /translation="MYRMQLLSCIALSLALVTNS"
     CDS             3597..3623
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             5898..5921
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
ORIGIN
        1 acaagagcag gtggcagcag gggaacgtct tctcatgctc cgtgatgcac gaggctctgc
       61 acaaccacta cacgcagaag agcctctccc tgtctccggg taaatgagtg ctagctggcc
      121 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
      181 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
      241 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg
      301 ggaggttttt taaagcaagt aaaacctcta caaatgtggt atggaattaa ttctaaaata
      361 cagcatagca aaactttaac ctccaaatca agcctctact tgaatccttt tctgagggat
      421 gaataaggca taggcatcag gggctgttgc caatgtgcat tagctgtttg cagcctcacc
      481 ttctttcatg gagtttaaga tatagtgtat tttcccaagg tttgaactag ctcttcattt
      541 ctttatgttt taaatgcact gacctcccac attccctttt tagtaaaata ttcagaaata
      601 atttaaatac atcattgcaa tgaaaataaa tgttttttat taggcagaat ccagatgctc
      661 aaggcccttc ataatatccc ccagtttagt agttggactt agggaacaaa ggaaccttta
      721 atagaaattg gacagcaaga aagcgagctt ctagcttatc ctcagtcctg ctcctctgcc
      781 acaaagtgca cgcagttgcc ggccgggtcg cgcagggcga actcccgccc ccacggctgc
      841 tcgccgatct cggtcatggc cggcccggag gcgtcccgga agttcgtgga cacgacctcc
      901 gaccactcgg cgtacagctc gtccaggccg cgcacccaca cccaggccag ggtgttgtcc
      961 ggcaccacct ggtcctggac cgcgctgatg aacagggtca cgtcgtcccg gaccacaccg
     1021 gcgaagtcgt cctccacgaa gtcccgggag aacccgagcc ggtcggtcca gaactcgacc
     1081 gctccggcga cgtcgcgcgc ggtgagcacc ggaacggcac tggtcaactt ggccatgatg
     1141 gctcctcctg tcaggagagg aaagagaaga aggttagtac aattgctata gtgagttgta
     1201 ttatactatg cagatatact atgccaatga ttaattgtca aactagggct gcagggttca
     1261 tagtgccact tttcctgcac tgccccatct cctgcccacc ctttcccagg catagacagt
     1321 cagtgactta ccaaactcac aggagggaga aggcagaagc ttgagacaga cccgcgggac
     1381 cgccgaactg cgaggggacg tggctagggc ggcttctttt atggtgcgcc ggccctcgga
     1441 ggcagggcgc tcggggaggc ctagcggcca atctgcggtg gcaggaggcg gggccgaagg
     1501 ccgtgcctga ccaatccgga gcacatagga gtctcagccc cccgccccaa agcaagggga
     1561 agtcacgcgc ctgtagcgcc agcgtgttgt gaaatggggg cttggggggg ttggggccct
     1621 gactagtcaa aacaaactcc cattgacgtc aatggggtgg agacttggaa atccccgtga
     1681 gtcaaaccgc tatccacgcc cattgatgta ctgccaaaac cgcatcatca tggtaatagc
     1741 gatgactaat acgtagatgt actgccaagt aggaaagtcc cataaggtca tgtactgggc
     1801 ataatgccag gcgggccatt taccgtcatt gacgtcaata gggggcgtac ttggcatatg
     1861 atacacttga tgtactgcca agtgggcagt ttaccgtaaa tactccaccc attgacgtca
     1921 atggaaagtc cctattggcg ttactatggg aacatacgtc attattgacg tcaatgggcg
     1981 ggggtcgttg ggcggtcagc caggcgggcc atttaccgta agttatgtaa cgcctgcagg
     2041 ttaattaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     2101 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     2161 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     2221 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     2281 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     2341 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     2401 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     2461 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     2521 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     2581 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     2641 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     2701 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     2761 gggattttgg tcatggctag ttaattaaca tttaaatcag cggccgcaat aaaatatctt
     2821 tattttcatt acatctgtgt gttggttttt tgtgtgaatc gtaactaaca tacgctctcc
     2881 atcaaaacaa aacgaaacaa aacaaactag caaaataggc tgtccccagt gcaagtgcag
     2941 gtgccagaac atttctctat cgaaggatct gcgatcgctc cggtgcccgt cagtgggcag
     3001 agcgcacatc gcccacagtc cccgagaagt tggggggagg ggtcggcaat tgaacgggtg
     3061 cctagagaag gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt
     3121 ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac gttctttttc
     3181 gcaacgggtt tgccgccaga acacagctga agcttcgagg ggctcgcatc tctccttcac
     3241 gcgcccgccg ccctacctga ggccgccatc cacgccggtt gagtcgcgtt ctgccgcctc
     3301 ccgcctgtgg tgcctcctga actgcgtccg ccgtctaggt aagtttaaag ctcaggtcga
     3361 gaccgggcct ttgtccggcg ctcccttgga gcctacctag actcagccgg ctctccacgc
     3421 tttgcctgac cctgcttgct caactctacg tctttgtttc gttttctgtt ctgcgccgtt
     3481 acagatccaa gctgtgaccg gcgcctacct gagatcaccg gcgaaggagg gccaccatgt
     3541 acaggatgca actcctgtct tgcattgcac taagtcttgc acttgtcacg aattcgtacc
     3601 catacgatgt tccagattac gctggatcca aaattgaaga aggcaaactg gtgatttgga
     3661 ttaacggcga taaaggctat aacggcctgg cggaagtggg caaaaaattt gaaaaagata
     3721 ccggcattaa agtgaccgtg gaacatccgg ataaactgga agaaaaattt ccgcaggtgg
     3781 cggcgaccgg cgatggcccg gatattattt tttgggcgca tgatcgcttt ggcggctatg
     3841 cgcagagcgg cctgctggcg gaaattaccc cggataaagc gtttcaggat aaactgtatc
     3901 cgtttacctg ggatgcggtg cgctataacg gcaaactgat tgcgtatccg attgcggtgg
     3961 aagcgctgag cctgatttat aacaaagatc tgctgccgaa cccgccgaaa acctgggaag
     4021 aaattccggc gctggataaa gaactgaaag cgaaaggcaa aagcgcgctg atgtttaacc
     4081 tgcaggaacc gtattttacc tggccgctga ttgcggcgga tggcggctat gcgtttaaat
     4141 atgaaaacgg caaatatgat attaaagatg tgggcgtgga taacgcgggc gcgaaagcgg
     4201 gcctgacctt tctggtggat ctgattaaaa acaaacatat gaacgcggat accgattata
     4261 gcattgcgga agcggcgttt aacaaaggcg aaaccgcgat gaccattaac ggcccgtggg
     4321 cgtggagcaa cattgatacc agcaaagtga actatggcgt gaccgtgctg ccgaccttta
     4381 aaggccagcc gagcaaaccg tttgtgggcg tgctgagcgc gggcattaac gcggcgagcc
     4441 cgaacaaaga actggcgaaa gaatttctgg aaaactatct gctgaccgat gaaggcctgg
     4501 aagcggtgaa caaagataaa ccgctgggcg cggtggcgct gaaaagctat gaagaagaac
     4561 tggcgaaaga tccgcgcatt gcggcgacca tggaaaacgc gcagaaaggc gaaattatgc
     4621 cgaacattcc gcagatgagc gcgttttggt atgcggtgcg caccgcggtg attaacgcgg
     4681 cgagcggccg ccagaccgtg gatgaagcgc tgaaagatgc gcagaccaac gagaaccttt
     4741 acttccaaac acactttgga gttctgatgg accttccaag aagtgcctca cagttagatg
     4801 caagaaacac taaagtcctc actttcatca gctatattgg gtgtggaata tctgctattt
     4861 tttcagcagc aactctcctg acatatgttg cttttgagaa attgcgaagg gattatccct
     4921 ccaaaatctt gatgaacctg agcacagccc tgctgttcct gaatctcctc ttcctcctag
     4981 atggctggat cacctccttc aatgtggatg gactttgcat tgctgttgca gtcctgttgc
     5041 atttcttcct tctggcaacc tttacctgga tggggctaga agcaattcac atgtacattg
     5101 ctctagttaa agtatttaac acttacattc gccgatacat tctaaaattc tgcatcattg
     5161 gctggggttt gcctgcctta gtggtgtcag ttgttctagc gagcagaaac aacaatgaag
     5221 tctatggaaa agaaagttat gggaaagaaa aaggtgatga attctgttgg attcaagatc
     5281 cagtcatatt ttatgtgacc tgtgctgggt attttggagt catgtttttt ctgaacattg
     5341 ccatgttcat tgtggtaatg gtgcagatct gtgggaggaa tggcaagaga agcaaccgga
     5401 ccctgagaga agaagtgtta aggaacctgc gcagtgtggt tagcttgacc tttctgttgg
     5461 gcatgacatg gggttttgca ttctttgcct ggggaccctt aaatatcccc ttcatgtacc
     5521 tcttctccat cttcaattca ttacaaggct tatttatatt catcttccac tgtgctatga
     5581 aggagaatgt tcagaaacag tggcggcagc atctctgctg tggtagattt cggttagcag
     5641 ataactcaga ttggagtaag acagctacca atatcatcaa gaaaagttct gataatctag
     5701 gaaaatcttt gtcttcaagc tccattggtt ccaactcaac ctatcttaca tccaaatcta
     5761 aatccagctc taccacctat ttcaaaagga atagccacac agataatgtc tcctatgagc
     5821 attccttcaa caaaagtgga tcactcagac agtgcttcca tggacaagtc cttgtcaaaa
     5881 ctggcccatg cggttctgac tacaaagacg atgacgacaa gtgaagatct gacaaaactc
     5941 acacatgccc accgtgccca gcacctgaac tcctgggggg accgtcagtc ttcctcttcc
     6001 ccccaaaacc caaggacacc ctcatgatct cccggacccc tgaggtcaca tgcgtggtgg
     6061 tggacgtgag ccacgaagac cctgaggtca agttcaactg gtacgtggac ggcgtggagg
     6121 tgcataatgc caagacaaag ccgcgggagg agcagtacaa cagcacgtac cgtgtggtca
     6181 gcgtcctcac cgtcctgcac caggactggc tgaatggcaa ggagtacaag tgcaaggtct
     6241 ccaacaaagc cctcccagcc cccatcgaga aaaccatctc caaagccaaa gggcagcccc
     6301 gagaaccaca ggtgtacacc ctgcccccat cccgggagga gatgaccaag aaccaggtca
     6361 gcctgacctg cctggtcaaa ggcttctatc ccagcgacat cgccgtggag tgggagagca
     6421 atgggcagcc ggagaacaac tacaagacca cgcctcccgt gctggactcc gacggctcct
     6481 tcttcctcta cagcaagctc accgtgg
//
