LOCUS       sequence-430280-        6282 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 6282)
  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 6282)
  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-ADGRE2 CTF-FLAG
FEATURES             Location/Qualifiers
     source          1..6282
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    16..137
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(27..46)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     81..100
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     polyA_signal    241..635
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     CDS             complement(649..1023)
                     /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        1616..1919
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(1999..2587)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2079..2098)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    2695..2743
                     /note="synthetic polyadenylation signal"
     misc_feature    2754..2845
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     2794..2813
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        2882..3093
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             3106..3374
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     sig_peptide     3424..3483
                     /codon_start=1
                     /product="human interleukin-2 signal sequence"
                     /label=IL-2 signal sequence
                     /translation="MYRMQLLSCIALSLALVTNS"
     CDS             3484..3510
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4618..4638
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             5560..5583
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
ORIGIN
        1 gctggccaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt
       61 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
      121 gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg
      181 aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtatg gaattaattc
      241 taaaatacag catagcaaaa ctttaacctc caaatcaagc ctctacttga atccttttct
      301 gagggatgaa taaggcatag gcatcagggg ctgttgccaa tgtgcattag ctgtttgcag
      361 cctcaccttc tttcatggag tttaagatat agtgtatttt cccaaggttt gaactagctc
      421 ttcatttctt tatgttttaa atgcactgac ctcccacatt ccctttttag taaaatattc
      481 agaaataatt taaatacatc attgcaatga aaataaatgt tttttattag gcagaatcca
      541 gatgctcaag gcccttcata atatccccca gtttagtagt tggacttagg gaacaaagga
      601 acctttaata gaaattggac agcaagaaag cgagcttcta gcttatcctc agtcctgctc
      661 ctctgccaca aagtgcacgc agttgccggc cgggtcgcgc agggcgaact cccgccccca
      721 cggctgctcg ccgatctcgg tcatggccgg cccggaggcg tcccggaagt tcgtggacac
      781 gacctccgac cactcggcgt acagctcgtc caggccgcgc acccacaccc aggccagggt
      841 gttgtccggc accacctggt cctggaccgc gctgatgaac agggtcacgt cgtcccggac
      901 cacaccggcg aagtcgtcct ccacgaagtc ccgggagaac ccgagccggt cggtccagaa
      961 ctcgaccgct ccggcgacgt cgcgcgcggt gagcaccgga acggcactgg tcaacttggc
     1021 catgatggct cctcctgtca ggagaggaaa gagaagaagg ttagtacaat tgctatagtg
     1081 agttgtatta tactatgcag atatactatg ccaatgatta attgtcaaac tagggctgca
     1141 gggttcatag tgccactttt cctgcactgc cccatctcct gcccaccctt tcccaggcat
     1201 agacagtcag tgacttacca aactcacagg agggagaagg cagaagcttg agacagaccc
     1261 gcgggaccgc cgaactgcga ggggacgtgg ctagggcggc ttcttttatg gtgcgccggc
     1321 cctcggaggc agggcgctcg gggaggccta gcggccaatc tgcggtggca ggaggcgggg
     1381 ccgaaggccg tgcctgacca atccggagca cataggagtc tcagcccccc gccccaaagc
     1441 aaggggaagt cacgcgcctg tagcgccagc gtgttgtgaa atgggggctt gggggggttg
     1501 gggccctgac tagtcaaaac aaactcccat tgacgtcaat ggggtggaga cttggaaatc
     1561 cccgtgagtc aaaccgctat ccacgcccat tgatgtactg ccaaaaccgc atcatcatgg
     1621 taatagcgat gactaatacg tagatgtact gccaagtagg aaagtcccat aaggtcatgt
     1681 actgggcata atgccaggcg ggccatttac cgtcattgac gtcaataggg ggcgtacttg
     1741 gcatatgata cacttgatgt actgccaagt gggcagttta ccgtaaatac tccacccatt
     1801 gacgtcaatg gaaagtccct attggcgtta ctatgggaac atacgtcatt attgacgtca
     1861 atgggcgggg gtcgttgggc ggtcagccag gcgggccatt taccgtaagt tatgtaacgc
     1921 ctgcaggtta attaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     1981 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     2041 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     2101 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     2161 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     2221 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     2281 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     2341 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     2401 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     2461 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     2521 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     2581 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     2641 acgttaaggg attttggtca tggctagtta attaacattt aaatcagcgg ccgcaataaa
     2701 atatctttat tttcattaca tctgtgtgtt ggttttttgt gtgaatcgta actaacatac
     2761 gctctccatc aaaacaaaac gaaacaaaac aaactagcaa aataggctgt ccccagtgca
     2821 agtgcaggtg ccagaacatt tctctatcga aggatctgcg atcgctccgg tgcccgtcag
     2881 tgggcagagc gcacatcgcc cacagtcccc gagaagttgg ggggaggggt cggcaattga
     2941 acgggtgcct agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt gtactggctc
     3001 cgcctttttc ccgagggtgg gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt
     3061 ctttttcgca acgggtttgc cgccagaaca cagctgaagc ttcgaggggc tcgcatctct
     3121 ccttcacgcg cccgccgccc tacctgaggc cgccatccac gccggttgag tcgcgttctg
     3181 ccgcctcccg cctgtggtgc ctcctgaact gcgtccgccg tctaggtaag tttaaagctc
     3241 aggtcgagac cgggcctttg tccggcgctc ccttggagcc tacctagact cagccggctc
     3301 tccacgcttt gcctgaccct gcttgctcaa ctctacgtct ttgtttcgtt ttctgttctg
     3361 cgccgttaca gatccaagct gtgaccggcg cctacctgag atcaccggcg aaggagggcc
     3421 accatgtaca ggatgcaact cctgtcttgc attgcactaa gtcttgcact tgtcacgaat
     3481 tcgtacccat acgatgttcc agattacgct ggatccaaaa ttgaagaagg caaactggtg
     3541 atttggatta acggcgataa aggctataac ggcctggcgg aagtgggcaa aaaatttgaa
     3601 aaagataccg gcattaaagt gaccgtggaa catccggata aactggaaga aaaatttccg
     3661 caggtggcgg cgaccggcga tggcccggat attatttttt gggcgcatga tcgctttggc
     3721 ggctatgcgc agagcggcct gctggcggaa attaccccgg ataaagcgtt tcaggataaa
     3781 ctgtatccgt ttacctggga tgcggtgcgc tataacggca aactgattgc gtatccgatt
     3841 gcggtggaag cgctgagcct gatttataac aaagatctgc tgccgaaccc gccgaaaacc
     3901 tgggaagaaa ttccggcgct ggataaagaa ctgaaagcga aaggcaaaag cgcgctgatg
     3961 tttaacctgc aggaaccgta ttttacctgg ccgctgattg cggcggatgg cggctatgcg
     4021 tttaaatatg aaaacggcaa atatgatatt aaagatgtgg gcgtggataa cgcgggcgcg
     4081 aaagcgggcc tgacctttct ggtggatctg attaaaaaca aacatatgaa cgcggatacc
     4141 gattatagca ttgcggaagc ggcgtttaac aaaggcgaaa ccgcgatgac cattaacggc
     4201 ccgtgggcgt ggagcaacat tgataccagc aaagtgaact atggcgtgac cgtgctgccg
     4261 acctttaaag gccagccgag caaaccgttt gtgggcgtgc tgagcgcggg cattaacgcg
     4321 gcgagcccga acaaagaact ggcgaaagaa tttctggaaa actatctgct gaccgatgaa
     4381 ggcctggaag cggtgaacaa agataaaccg ctgggcgcgg tggcgctgaa aagctatgaa
     4441 gaagaactgg cgaaagatcc gcgcattgcg gcgaccatgg aaaacgcgca gaaaggcgaa
     4501 attatgccga acattccgca gatgagcgcg ttttggtatg cggtgcgcac cgcggtgatt
     4561 aacgcggcga gcggccgcca gaccgtggat gaagcgctga aagatgcgca gaccaacgag
     4621 aacctttact tccaaagcag ctttgccgtc ctcatggccc actacgatgt gcaggaggag
     4681 gatcccgtgc tgactgtcat cacctacatg gggctgagcg tctctctgct gtgcctcctc
     4741 ctggcggccc tcacttttct cctgtgtaaa gccatccaga acaccagcac ctcactgcat
     4801 ctgcagctct cgctctgcct cttcctggcc cacctcctct tcctcgtggc aattgatcaa
     4861 accggacaca aggtgctgtg ctccatcatc gccggtacct tgcactatct ctacctggcc
     4921 accttgacct ggatgctgct ggaggccctg tacctcttcc tcactgcacg gaacctgacg
     4981 gtggtcaact actcaagcat caacagattc atgaagaagc tcatgttccc tgtgggctac
     5041 ggagtcccag ctgtgacagt ggccatttct gcagcctcca ggcctcacct ttatggaaca
     5101 ccttcccgct gctggctcca accagaaaag ggatttatat ggggcttcct tggacctgtc
     5161 tgcgccatct tctctgtgaa tttagttctc tttctggtga ctctctggat tttgaaaaac
     5221 agactctcct ccctcaatag tgaagtgtcc accctccgga acacaaggat gctggcattt
     5281 aaagcgacag ctcagctgtt catcctgggc tgcacgtggt gtctgggcat cttgcaggtg
     5341 ggtccggctg cccgggtcat ggcctacctc ttcaccatca tcaacagcct gcagggtgtc
     5401 ttcatcttcc tggtgtactg cctcctcagc cagcaggtcc gggagcaata tgggaaatgg
     5461 tccaaaggga tcaggaaatt gaaaactgag tctgagatgc acacactctc cagcagtgct
     5521 aaggctgaca cctccaaacc cagcacggtt aacggttctg actacaaaga cgatgacgac
     5581 aagtgaagat ctgacaaaac tcacacatgc ccaccgtgcc cagcacctga actcctgggg
     5641 ggaccgtcag tcttcctctt ccccccaaaa cccaaggaca ccctcatgat ctcccggacc
     5701 cctgaggtca catgcgtggt ggtggacgtg agccacgaag accctgaggt caagttcaac
     5761 tggtacgtgg acggcgtgga ggtgcataat gccaagacaa agccgcggga ggagcagtac
     5821 aacagcacgt accgtgtggt cagcgtcctc accgtcctgc accaggactg gctgaatggc
     5881 aaggagtaca agtgcaaggt ctccaacaaa gccctcccag cccccatcga gaaaaccatc
     5941 tccaaagcca aagggcagcc ccgagaacca caggtgtaca ccctgccccc atcccgggag
     6001 gagatgacca agaaccaggt cagcctgacc tgcctggtca aaggcttcta tcccagcgac
     6061 atcgccgtgg agtgggagag caatgggcag ccggagaaca actacaagac cacgcctccc
     6121 gtgctggact ccgacggctc cttcttcctc tacagcaagc tcaccgtgga caagagcagg
     6181 tggcagcagg ggaacgtctt ctcatgctcc gtgatgcacg aggctctgca caaccactac
     6241 acgcagaaga gcctctccct gtctccgggt aaatgagtgc ta
//
