LOCUS       sequence-442648-        6510 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 6510)
  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 6510)
  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-ADGRG7 CTF-FLAG
FEATURES             Location/Qualifiers
     source          1..6510
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1167..1193)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     sig_peptide     complement(1194..1253)
                     /codon_start=1
                     /product="human interleukin-2 signal sequence"
                     /label=IL-2 signal sequence
                     /translation="MYRMQLLSCIALSLALVTNS"
     LTR             1303..1571
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     promoter        complement(1584..1795)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     misc_feature    1832..1923
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     complement(1864..1883)
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     polyA_signal    1934..1982
                     /note="synthetic polyadenylation signal"
     rep_origin      2090..2678
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2579..2598
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2758..3061
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             3654..4028
                     /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    4042..4436
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     polyA_signal    4540..4661
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4577..4596)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4631..4650
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(5376..5399)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
ORIGIN
        1 tggtagtctt tcttgaatgt cataagcact gcaaagttgg tttggaagta aaggttctcg
       61 ttggtctgcg catctttcag cgcttcatcc acggtctggc ggccgctcgc cgcgttaatc
      121 accgcggtgc gcaccgcata ccaaaacgcg ctcatctgcg gaatgttcgg cataatttcg
      181 cctttctgcg cgttttccat ggtcgccgca atgcgcggat ctttcgccag ttcttcttca
      241 tagcttttca gcgccaccgc gcccagcggt ttatctttgt tcaccgcttc caggccttca
      301 tcggtcagca gatagttttc cagaaattct ttcgccagtt ctttgttcgg gctcgccgcg
      361 ttaatgcccg cgctcagcac gcccacaaac ggtttgctcg gctggccttt aaaggtcggc
      421 agcacggtca cgccatagtt cactttgctg gtatcaatgt tgctccacgc ccacgggccg
      481 ttaatggtca tcgcggtttc gcctttgtta aacgccgctt ccgcaatgct ataatcggta
      541 tccgcgttca tatgtttgtt tttaatcaga tccaccagaa aggtcaggcc cgctttcgcg
      601 cccgcgttat ccacgcccac atctttaata tcatatttgc cgttttcata tttaaacgca
      661 tagccgccat ccgccgcaat cagcggccag gtaaaatacg gttcctgcag gttaaacatc
      721 agcgcgcttt tgcctttcgc tttcagttct ttatccagcg ccggaatttc ttcccaggtt
      781 ttcggcgggt tcggcagcag atctttgtta taaatcaggc tcagcgcttc caccgcaatc
      841 ggatacgcaa tcagtttgcc gttatagcgc accgcatccc aggtaaacgg atacagttta
      901 tcctgaaacg ctttatccgg ggtaatttcc gccagcaggc cgctctgcgc atagccgcca
      961 aagcgatcat gcgcccaaaa aataatatcc gggccatcgc cggtcgccgc cacctgcgga
     1021 aatttttctt ccagtttatc cggatgttcc acggtcactt taatgccggt atctttttca
     1081 aattttttgc ccacttccgc caggccgtta tagcctttat cgccgttaat ccaaatcacc
     1141 agtttgcctt cttcaatttt ggatccagcg taatctggaa catcgtatgg gtacgaattc
     1201 gtgacaagtg caagacttag tgcaatgcaa gacaggagtt gcatcctgta catggtggcc
     1261 ctccttcgcc ggtgatctca ggtaggcgcc ggtcacagct tggatctgta acggcgcaga
     1321 acagaaaacg aaacaaagac gtagagttga gcaagcaggg tcaggcaaag cgtggagagc
     1381 cggctgagtc taggtaggct ccaagggagc gccggacaaa ggcccggtct cgacctgagc
     1441 tttaaactta cctagacggc ggacgcagtt caggaggcac cacaggcggg aggcggcaga
     1501 acgcgactca accggcgtgg atggcggcct caggtagggc ggcgggcgcg tgaaggagag
     1561 atgcgagccc ctcgaagctt cagctgtgtt ctggcggcaa acccgttgcg aaaaagaacg
     1621 ttcacggcga ctactgcact tatatacggt tctcccccac cctcgggaaa aaggcggagc
     1681 cagtacacga catcactttc ccagtttacc ccgcgccacc ttctctaggc acccgttcaa
     1741 ttgccgaccc ctccccccaa cttctcgggg actgtgggcg atgtgcgctc tgcccactga
     1801 cgggcaccgg agcgatcgca gatccttcga tagagaaatg ttctggcacc tgcacttgca
     1861 ctggggacag cctattttgc tagtttgttt tgtttcgttt tgttttgatg gagagcgtat
     1921 gttagttacg attcacacaa aaaaccaaca cacagatgta atgaaaataa agatatttta
     1981 ttgcggccgc tgatttaaat gttaattaac tagccatgac caaaatccct taacgtgagt
     2041 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     2101 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     2161 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     2221 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     2281 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     2341 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     2401 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     2461 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     2521 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     2581 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     2641 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     2701 tacggttcct ggccttttgc tggccttttg ctcacatgtt cttaattaac ctgcaggcgt
     2761 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
     2821 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
     2881 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
     2941 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
     3001 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
     3061 gatgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
     3121 tccaagtctc caccccattg acgtcaatgg gagtttgttt tgactagtca gggccccaac
     3181 ccccccaagc ccccatttca caacacgctg gcgctacagg cgcgtgactt ccccttgctt
     3241 tggggcgggg ggctgagact cctatgtgct ccggattggt caggcacggc cttcggcccc
     3301 gcctcctgcc accgcagatt ggccgctagg cctccccgag cgccctgcct ccgagggccg
     3361 gcgcaccata aaagaagccg ccctagccac gtcccctcgc agttcggcgg tcccgcgggt
     3421 ctgtctcaag cttctgcctt ctccctcctg tgagtttggt aagtcactga ctgtctatgc
     3481 ctgggaaagg gtgggcagga gatggggcag tgcaggaaaa gtggcactat gaaccctgca
     3541 gccctagttt gacaattaat cattggcata gtatatctgc atagtataat acaactcact
     3601 atagcaattg tactaacctt cttctctttc ctctcctgac aggaggagcc atcatggcca
     3661 agttgaccag tgccgttccg gtgctcaccg cgcgcgacgt cgccggagcg gtcgagttct
     3721 ggaccgaccg gctcgggttc tcccgggact tcgtggagga cgacttcgcc ggtgtggtcc
     3781 gggacgacgt gaccctgttc atcagcgcgg tccaggacca ggtggtgccg gacaacaccc
     3841 tggcctgggt gtgggtgcgc ggcctggacg agctgtacgc cgagtggtcg gaggtcgtgt
     3901 ccacgaactt ccgggacgcc tccgggccgg ccatgaccga gatcggcgag cagccgtggg
     3961 ggcgggagtt cgccctgcgc gacccggccg gcaactgcgt gcactttgtg gcagaggagc
     4021 aggactgagg ataagctaga agctcgcttt cttgctgtcc aatttctatt aaaggttcct
     4081 ttgttcccta agtccaacta ctaaactggg ggatattatg aagggccttg agcatctgga
     4141 ttctgcctaa taaaaaacat ttattttcat tgcaatgatg tatttaaatt atttctgaat
     4201 attttactaa aaagggaatg tgggaggtca gtgcatttaa aacataaaga aatgaagagc
     4261 tagttcaaac cttgggaaaa tacactatat cttaaactcc atgaaagaag gtgaggctgc
     4321 aaacagctaa tgcacattgg caacagcccc tgatgcctat gccttattca tccctcagaa
     4381 aaggattcaa gtagaggctt gatttggagg ttaaagtttt gctatgctgt attttagaat
     4441 taattccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc
     4501 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat
     4561 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     4621 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg gccagctagc
     4681 actcatttac ccggagacag ggagaggctc ttctgcgtgt agtggttgtg cagagcctcg
     4741 tgcatcacgg agcatgagaa gacgttcccc tgctgccacc tgctcttgtc cacggtgagc
     4801 ttgctgtaga ggaagaagga gccgtcggag tccagcacgg gaggcgtggt cttgtagttg
     4861 ttctccggct gcccattgct ctcccactcc acggcgatgt cgctgggata gaagcctttg
     4921 accaggcagg tcaggctgac ctggttcttg gtcatctcct cccgggatgg gggcagggtg
     4981 tacacctgtg gttctcgggg ctgccctttg gctttggaga tggttttctc gatgggggct
     5041 gggagggctt tgttggagac cttgcacttg tactccttgc cattcagcca gtcctggtgc
     5101 aggacggtga ggacgctgac cacacggtac gtgctgttgt actgctcctc ccgcggcttt
     5161 gtcttggcat tatgcacctc cacgccgtcc acgtaccagt tgaacttgac ctcagggtct
     5221 tcgtggctca cgtccaccac cacgcatgtg acctcagggg tccgggagat catgagggtg
     5281 tccttgggtt ttggggggaa gaggaagact gacggtcccc ccaggagttc aggtgctggg
     5341 cacggtgggc atgtgtgagt tttgtcagat cttcacttgt cgtcatcgtc tttgtagtca
     5401 gaaccgatag actccttagc gttgtcactt tcgctcaggg taatctcttc agtggaggga
     5461 ctggtctcca agaggcgaaa gcgctcatgc agtgtaggca agctgcggag aaagttgtac
     5521 atcttgacgc gcaggcgggg tctagtcacg ctgggcaggc ttttgcgcct accaatggaa
     5581 ctcaggagca tcaaaacctt gcttgcttcg ctctggaaga cctttgtgcg cacagtgtaa
     5641 aggataaaga tctgcaggcc ctgggttgtg ttgaaaaggc agaaaatgta ggagaacacg
     5701 atgcggatag aatcgtcatt caccagcatc agataggcga ggatccatgt gatgccgaag
     5761 acaaccgcca cagacagtgt ggacacaatc tttttcatag aggacacctt tttagtgcta
     5821 gtcaagttct ggttattctt ccagagcact ttgatgctga ttgtgatgaa catgaccacg
     5881 ttgctaatca gaatgatggt cacaggcacg atgaatgacc acaggagagg actcttgatc
     5941 accccgttag gttccgggat tgccagccag cagatttttt cctgacggta gtccagctcc
     6001 cattgaggat tgtttccgtt ttgagagtag attactccga cagtaattgc cactacgata
     6061 gcgggtactc cccatccgat cagactaatg aacaggatga aatggcgcgg cagaggcttc
     6121 attgtgcgga tcagaaggta atacaactga gctgcggaaa gagcgttcca agtgaaggtt
     6181 accagaagaa aatagtggag cagagcggcg atcgccgtac acatggggtt ggggatatta
     6241 attgtatcgg tcctgggaat atcattgttg tcgaaatcaa tgttattgat atctccatcg
     6301 gaggtctgca ggttcttatt ggaattctcg atcccaaaca cgaacagcaa gttgaagatc
     6361 agcatagaga tgcagaggtt gacgagaacc catgtcacgg aggtctttct caccttgcgg
     6421 gtaacgatct gaaaaataac cgtcaacgcg agccctgtga cgctcagggc acaaccgacg
     6481 ttagacagga tgtcaagact tttagggtat
//
