LOCUS       sequence-430283-        6396 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 6396)
  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 6396)
  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-ADGRF1 CTF-FLAG
FEATURES             Location/Qualifiers
     source          1..6396
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(644..667)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(2831..2857)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     sig_peptide     complement(2858..2917)
                     /codon_start=1
                     /product="human interleukin-2 signal sequence"
                     /label=IL-2 signal sequence
                     /translation="MYRMQLLSCIALSLALVTNS"
     LTR             2967..3235
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     promoter        complement(3248..3459)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     misc_feature    3496..3587
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     complement(3528..3547)
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     polyA_signal    3598..3646
                     /note="synthetic polyadenylation signal"
     rep_origin      3754..4342
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4243..4262
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        4422..4725
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             5318..5692
                     /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"
     polyA_signal    5706..6100
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     polyA_signal    6204..6325
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6241..6260)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6295..6314
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 gagcctcgtg catcacggag catgagaaga cgttcccctg ctgccacctg ctcttgtcca
       61 cggtgagctt gctgtagagg aagaaggagc cgtcggagtc cagcacggga ggcgtggtct
      121 tgtagttgtt ctccggctgc ccattgctct cccactccac ggcgatgtcg ctgggataga
      181 agcctttgac caggcaggtc aggctgacct ggttcttggt catctcctcc cgggatgggg
      241 gcagggtgta cacctgtggt tctcggggct gccctttggc tttggagatg gttttctcga
      301 tgggggctgg gagggctttg ttggagacct tgcacttgta ctccttgcca ttcagccagt
      361 cctggtgcag gacggtgagg acgctgacca cacggtacgt gctgttgtac tgctcctccc
      421 gcggctttgt cttggcatta tgcacctcca cgccgtccac gtaccagttg aacttgacct
      481 cagggtcttc gtggctcacg tccaccacca cgcatgtgac ctcaggggtc cgggagatca
      541 tgagggtgtc cttgggtttt ggggggaaga ggaagactga cggtcccccc aggagttcag
      601 gtgctgggca cggtgggcat gtgtgagttt tgtcagatct tcacttgtcg tcatcgtctt
      661 tgtagtcaga accttcattt gagacaaact gagttagcat gatgttgtcg gaggaatctc
      721 cagtatgaga aaatgcataa tggcctttgt tttgcagtgg gttgaaaggc tttgagaatt
      781 tgggtttggc agataaatct gatgagtttt gcttttctgt ttgcttccaa gaacttaagg
      841 cagacaactt gttgaacaga agttgtcgca gcttactgtc caagagtatt ccaaagcata
      901 agataaaaaa tccctggaat gcattgagta aagcaaaaat aacatgccaa gccagattct
      961 ggctgtccac tattgttcct attccaaagc cccaggtgag ccctagcaga ggggtcagaa
     1021 tgaggaggct cttccccacg cggatgatgg tggccttgtc atcccgactc agtctttccc
     1081 caacagtcgg cctccagagc tttgtgagaa ctagcagcac cacaacgaag ttcacagcca
     1141 caatagccag tgcagggaca acaaaagcca ggagtggttt gcttccattg gaccagttaa
     1201 gccaacacac atctttcctt ttgtaggtat tgctaggttg tgtgacagca atggtaatga
     1261 cagatataat gagagggcac ccataaccca ggcaaaatcc aacagccatc atcaaatgct
     1321 gggccatgtg atggaacacg aggatgatcc ggtaagccag caggatgcca agcatgagca
     1381 tccagaagaa caaagagagg tagaagaagt gtgtaaagaa cacagcagct gtgcagactc
     1441 cagaagggtt caccgtggtg tccactgtgg caccaacaat aaaccagaca tcagcaatca
     1501 agagggacag ggctatgttc accatgcaaa tacgacgtgt gtgagaggtt tggctttttt
     1561 taatctgctt ccaaaacaaa gcctcgatga tcaggcataa aatgagactt ccaatggaga
     1621 tacccagtcc cacataggtg atccatttta caacggggaa gattgtagag gggacaaaag
     1681 gtgacatcaa tatggagaag gaggtttgga agtaaaggtt ctcgttggtc tgcgcatctt
     1741 tcagcgcttc atccacggtc tggcggccgc tcgccgcgtt aatcaccgcg gtgcgcaccg
     1801 cataccaaaa cgcgctcatc tgcggaatgt tcggcataat ttcgcctttc tgcgcgtttt
     1861 ccatggtcgc cgcaatgcgc ggatctttcg ccagttcttc ttcatagctt ttcagcgcca
     1921 ccgcgcccag cggtttatct ttgttcaccg cttccaggcc ttcatcggtc agcagatagt
     1981 tttccagaaa ttctttcgcc agttctttgt tcgggctcgc cgcgttaatg cccgcgctca
     2041 gcacgcccac aaacggtttg ctcggctggc ctttaaaggt cggcagcacg gtcacgccat
     2101 agttcacttt gctggtatca atgttgctcc acgcccacgg gccgttaatg gtcatcgcgg
     2161 tttcgccttt gttaaacgcc gcttccgcaa tgctataatc ggtatccgcg ttcatatgtt
     2221 tgtttttaat cagatccacc agaaaggtca ggcccgcttt cgcgcccgcg ttatccacgc
     2281 ccacatcttt aatatcatat ttgccgtttt catatttaaa cgcatagccg ccatccgccg
     2341 caatcagcgg ccaggtaaaa tacggttcct gcaggttaaa catcagcgcg cttttgcctt
     2401 tcgctttcag ttctttatcc agcgccggaa tttcttccca ggttttcggc gggttcggca
     2461 gcagatcttt gttataaatc aggctcagcg cttccaccgc aatcggatac gcaatcagtt
     2521 tgccgttata gcgcaccgca tcccaggtaa acggatacag tttatcctga aacgctttat
     2581 ccggggtaat ttccgccagc aggccgctct gcgcatagcc gccaaagcga tcatgcgccc
     2641 aaaaaataat atccgggcca tcgccggtcg ccgccacctg cggaaatttt tcttccagtt
     2701 tatccggatg ttccacggtc actttaatgc cggtatcttt ttcaaatttt ttgcccactt
     2761 ccgccaggcc gttatagcct ttatcgccgt taatccaaat caccagtttg ccttcttcaa
     2821 ttttggatcc agcgtaatct ggaacatcgt atgggtacga attcgtgaca agtgcaagac
     2881 ttagtgcaat gcaagacagg agttgcatcc tgtacatggt ggccctcctt cgccggtgat
     2941 ctcaggtagg cgccggtcac agcttggatc tgtaacggcg cagaacagaa aacgaaacaa
     3001 agacgtagag ttgagcaagc agggtcaggc aaagcgtgga gagccggctg agtctaggta
     3061 ggctccaagg gagcgccgga caaaggcccg gtctcgacct gagctttaaa cttacctaga
     3121 cggcggacgc agttcaggag gcaccacagg cgggaggcgg cagaacgcga ctcaaccggc
     3181 gtggatggcg gcctcaggta gggcggcggg cgcgtgaagg agagatgcga gcccctcgaa
     3241 gcttcagctg tgttctggcg gcaaacccgt tgcgaaaaag aacgttcacg gcgactactg
     3301 cacttatata cggttctccc ccaccctcgg gaaaaaggcg gagccagtac acgacatcac
     3361 tttcccagtt taccccgcgc caccttctct aggcacccgt tcaattgccg acccctcccc
     3421 ccaacttctc ggggactgtg ggcgatgtgc gctctgccca ctgacgggca ccggagcgat
     3481 cgcagatcct tcgatagaga aatgttctgg cacctgcact tgcactgggg acagcctatt
     3541 ttgctagttt gttttgtttc gttttgtttt gatggagagc gtatgttagt tacgattcac
     3601 acaaaaaacc aacacacaga tgtaatgaaa ataaagatat tttattgcgg ccgctgattt
     3661 aaatgttaat taactagcca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     3721 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     3781 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     3841 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     3901 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     3961 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     4021 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     4081 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     4141 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     4201 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     4261 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     4321 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     4381 ttgctggcct tttgctcaca tgttcttaat taacctgcag gcgttacata acttacggta
     4441 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat
     4501 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
     4561 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac
     4621 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt
     4681 cctacttggc agtacatcta cgtattagtc atcgctatta ccatgatgat gcggttttgg
     4741 cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc
     4801 attgacgtca atgggagttt gttttgacta gtcagggccc caaccccccc aagcccccat
     4861 ttcacaacac gctggcgcta caggcgcgtg acttcccctt gctttggggc ggggggctga
     4921 gactcctatg tgctccggat tggtcaggca cggccttcgg ccccgcctcc tgccaccgca
     4981 gattggccgc taggcctccc cgagcgccct gcctccgagg gccggcgcac cataaaagaa
     5041 gccgccctag ccacgtcccc tcgcagttcg gcggtcccgc gggtctgtct caagcttctg
     5101 ccttctccct cctgtgagtt tggtaagtca ctgactgtct atgcctggga aagggtgggc
     5161 aggagatggg gcagtgcagg aaaagtggca ctatgaaccc tgcagcccta gtttgacaat
     5221 taatcattgg catagtatat ctgcatagta taatacaact cactatagca attgtactaa
     5281 ccttcttctc tttcctctcc tgacaggagg agccatcatg gccaagttga ccagtgccgt
     5341 tccggtgctc accgcgcgcg acgtcgccgg agcggtcgag ttctggaccg accggctcgg
     5401 gttctcccgg gacttcgtgg aggacgactt cgccggtgtg gtccgggacg acgtgaccct
     5461 gttcatcagc gcggtccagg accaggtggt gccggacaac accctggcct gggtgtgggt
     5521 gcgcggcctg gacgagctgt acgccgagtg gtcggaggtc gtgtccacga acttccggga
     5581 cgcctccggg ccggccatga ccgagatcgg cgagcagccg tgggggcggg agttcgccct
     5641 gcgcgacccg gccggcaact gcgtgcactt tgtggcagag gagcaggact gaggataagc
     5701 tagaagctcg ctttcttgct gtccaatttc tattaaaggt tcctttgttc cctaagtcca
     5761 actactaaac tgggggatat tatgaagggc cttgagcatc tggattctgc ctaataaaaa
     5821 acatttattt tcattgcaat gatgtattta aattatttct gaatatttta ctaaaaaggg
     5881 aatgtgggag gtcagtgcat ttaaaacata aagaaatgaa gagctagttc aaaccttggg
     5941 aaaatacact atatcttaaa ctccatgaaa gaaggtgagg ctgcaaacag ctaatgcaca
     6001 ttggcaacag cccctgatgc ctatgcctta ttcatccctc agaaaaggat tcaagtagag
     6061 gcttgatttg gaggttaaag ttttgctatg ctgtatttta gaattaattc cataccacat
     6121 ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata
     6181 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
     6241 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     6301 tgtccaaact catcaatgta tcttatcatg tctggccagc tagcactcat ttacccggag
     6361 acagggagag gctcttctgc gtgtagtggt tgtgca
//
