LOCUS       sequence-430288-        6441 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 6441)
  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 6441)
  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-ADGRG4 CTF-FLAG
FEATURES             Location/Qualifiers
     source          1..6441
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    96..217
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(107..126)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     161..180
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     polyA_signal    321..715
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     CDS             complement(729..1103)
                     /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        1696..1999
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(2079..2667)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2159..2178)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    2775..2823
                     /note="synthetic polyadenylation signal"
     misc_feature    2834..2925
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     2874..2893
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        2962..3173
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             3186..3454
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     sig_peptide     3504..3563
                     /codon_start=1
                     /product="human interleukin-2 signal sequence"
                     /label=IL-2 signal sequence
                     /translation="MYRMQLLSCIALSLALVTNS"
     CDS             3564..3590
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             5799..5822
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
ORIGIN
        1 catgctccgt gatgcacgag gctctgcaca accactacac gcagaagagc ctctccctgt
       61 ctccgggtaa atgagtgcta gctggccaga catgataaga tacattgatg agtttggaca
      121 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
      181 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
      241 tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa
      301 atgtggtatg gaattaattc taaaatacag catagcaaaa ctttaacctc caaatcaagc
      361 ctctacttga atccttttct gagggatgaa taaggcatag gcatcagggg ctgttgccaa
      421 tgtgcattag ctgtttgcag cctcaccttc tttcatggag tttaagatat agtgtatttt
      481 cccaaggttt gaactagctc ttcatttctt tatgttttaa atgcactgac ctcccacatt
      541 ccctttttag taaaatattc agaaataatt taaatacatc attgcaatga aaataaatgt
      601 tttttattag gcagaatcca gatgctcaag gcccttcata atatccccca gtttagtagt
      661 tggacttagg gaacaaagga acctttaata gaaattggac agcaagaaag cgagcttcta
      721 gcttatcctc agtcctgctc ctctgccaca aagtgcacgc agttgccggc cgggtcgcgc
      781 agggcgaact cccgccccca cggctgctcg ccgatctcgg tcatggccgg cccggaggcg
      841 tcccggaagt tcgtggacac gacctccgac cactcggcgt acagctcgtc caggccgcgc
      901 acccacaccc aggccagggt gttgtccggc accacctggt cctggaccgc gctgatgaac
      961 agggtcacgt cgtcccggac cacaccggcg aagtcgtcct ccacgaagtc ccgggagaac
     1021 ccgagccggt cggtccagaa ctcgaccgct ccggcgacgt cgcgcgcggt gagcaccgga
     1081 acggcactgg tcaacttggc catgatggct cctcctgtca ggagaggaaa gagaagaagg
     1141 ttagtacaat tgctatagtg agttgtatta tactatgcag atatactatg ccaatgatta
     1201 attgtcaaac tagggctgca gggttcatag tgccactttt cctgcactgc cccatctcct
     1261 gcccaccctt tcccaggcat agacagtcag tgacttacca aactcacagg agggagaagg
     1321 cagaagcttg agacagaccc gcgggaccgc cgaactgcga ggggacgtgg ctagggcggc
     1381 ttcttttatg gtgcgccggc cctcggaggc agggcgctcg gggaggccta gcggccaatc
     1441 tgcggtggca ggaggcgggg ccgaaggccg tgcctgacca atccggagca cataggagtc
     1501 tcagcccccc gccccaaagc aaggggaagt cacgcgcctg tagcgccagc gtgttgtgaa
     1561 atgggggctt gggggggttg gggccctgac tagtcaaaac aaactcccat tgacgtcaat
     1621 ggggtggaga cttggaaatc cccgtgagtc aaaccgctat ccacgcccat tgatgtactg
     1681 ccaaaaccgc atcatcatgg taatagcgat gactaatacg tagatgtact gccaagtagg
     1741 aaagtcccat aaggtcatgt actgggcata atgccaggcg ggccatttac cgtcattgac
     1801 gtcaataggg ggcgtacttg gcatatgata cacttgatgt actgccaagt gggcagttta
     1861 ccgtaaatac tccacccatt gacgtcaatg gaaagtccct attggcgtta ctatgggaac
     1921 atacgtcatt attgacgtca atgggcgggg gtcgttgggc ggtcagccag gcgggccatt
     1981 taccgtaagt tatgtaacgc ctgcaggtta attaagaaca tgtgagcaaa aggccagcaa
     2041 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     2101 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     2161 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     2221 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     2281 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     2341 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     2401 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     2461 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     2521 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     2581 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     2641 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     2701 gctcagtgga acgaaaactc acgttaaggg attttggtca tggctagtta attaacattt
     2761 aaatcagcgg ccgcaataaa atatctttat tttcattaca tctgtgtgtt ggttttttgt
     2821 gtgaatcgta actaacatac gctctccatc aaaacaaaac gaaacaaaac aaactagcaa
     2881 aataggctgt ccccagtgca agtgcaggtg ccagaacatt tctctatcga aggatctgcg
     2941 atcgctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg
     3001 ggggaggggt cggcaattga acgggtgcct agagaaggtg gcgcggggta aactgggaaa
     3061 gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg
     3121 cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca cagctgaagc
     3181 ttcgaggggc tcgcatctct ccttcacgcg cccgccgccc tacctgaggc cgccatccac
     3241 gccggttgag tcgcgttctg ccgcctcccg cctgtggtgc ctcctgaact gcgtccgccg
     3301 tctaggtaag tttaaagctc aggtcgagac cgggcctttg tccggcgctc ccttggagcc
     3361 tacctagact cagccggctc tccacgcttt gcctgaccct gcttgctcaa ctctacgtct
     3421 ttgtttcgtt ttctgttctg cgccgttaca gatccaagct gtgaccggcg cctacctgag
     3481 atcaccggcg aaggagggcc accatgtaca ggatgcaact cctgtcttgc attgcactaa
     3541 gtcttgcact tgtcacgaat tcgtacccat acgatgttcc agattacgct ggatccaaaa
     3601 ttgaagaagg caaactggtg atttggatta acggcgataa aggctataac ggcctggcgg
     3661 aagtgggcaa aaaatttgaa aaagataccg gcattaaagt gaccgtggaa catccggata
     3721 aactggaaga aaaatttccg caggtggcgg cgaccggcga tggcccggat attatttttt
     3781 gggcgcatga tcgctttggc ggctatgcgc agagcggcct gctggcggaa attaccccgg
     3841 ataaagcgtt tcaggataaa ctgtatccgt ttacctggga tgcggtgcgc tataacggca
     3901 aactgattgc gtatccgatt gcggtggaag cgctgagcct gatttataac aaagatctgc
     3961 tgccgaaccc gccgaaaacc tgggaagaaa ttccggcgct ggataaagaa ctgaaagcga
     4021 aaggcaaaag cgcgctgatg tttaacctgc aggaaccgta ttttacctgg ccgctgattg
     4081 cggcggatgg cggctatgcg tttaaatatg aaaacggcaa atatgatatt aaagatgtgg
     4141 gcgtggataa cgcgggcgcg aaagcgggcc tgacctttct ggtggatctg attaaaaaca
     4201 aacatatgaa cgcggatacc gattatagca ttgcggaagc ggcgtttaac aaaggcgaaa
     4261 ccgcgatgac cattaacggc ccgtgggcgt ggagcaacat tgataccagc aaagtgaact
     4321 atggcgtgac cgtgctgccg acctttaaag gccagccgag caaaccgttt gtgggcgtgc
     4381 tgagcgcggg cattaacgcg gcgagcccga acaaagaact ggcgaaagaa tttctggaaa
     4441 actatctgct gaccgatgaa ggcctggaag cggtgaacaa agataaaccg ctgggcgcgg
     4501 tggcgctgaa aagctatgaa gaagaactgg cgaaagatcc gcgcattgcg gcgaccatgg
     4561 aaaacgcgca gaaaggcgaa attatgccga acattccgca gatgagcgcg ttttggtatg
     4621 cggtgcgcac cgcggtgatt aacgcggcga gcggccgcca gaccgtggat gaagcgctga
     4681 aagatgcgca gaccaacgag aacctttact tccaaaccca ctttggcgtg ctcatggacc
     4741 tctcaagatc taccgtggac tcagtgaatg agcagatcct ggccctgatc acctacaccg
     4801 ggtgtggcat cagtagcatt ttccttggcg tcgctgtagt gacatacatt gctttccaca
     4861 agctgcgcaa ggattaccct gcgaaaatcc tgatcaacct ctgtacagct ctgctcatgc
     4921 tgaacctggt cttcttgatc aactcctggc tctccagctt ccagaaggtg ggagtctgta
     4981 tcacagctgc agttgccctg cactatttct tgcttgtctc atttacatgg atgggacttg
     5041 aggccgtgca catgtacctg gccctcgtca aggttttcaa catctacatc ccgaactaca
     5101 tcctcaagtt ctgtctggtg ggctggggaa ttcccgcaat catggtggca attacggtct
     5161 ccgttaaaaa ggatctgtac ggcaccctct ccccgacaac cccgttttgt tggatcaagg
     5221 atgactccat cttttatatc agcgttgtgg cctatttttg cctgattttt ctgatgaacc
     5281 tgtctatgtt ttgcacggtc ctggtgcagc tgaattctgt aaagtcccag atccaaaaaa
     5341 ctcgccgtaa aatgatcctg catgacctga aaggcaccat gtccctcact tttctgctcg
     5401 gcctgacatg gggatttgcc ttctttgcct ggggaccgat gagaaatttc tttttgtatc
     5461 tttttgcaat ctttaacaca ctccagggat ttttcatctt tgtattccat tgtgttatga
     5521 aggagtcagt cagagagcaa tggcagatcc acttgtgctg tggctggctg cgtctcgata
     5581 actcctcaga cggaagctcc cgctgtcaga ttaaggttgg ctataagcag gaaggtctga
     5641 aaaagatttt tgagcacaaa ctgctcaccc cgtctctgaa atctaccgca acctcatcca
     5701 ccttcaaatc cctgggcagc gcccaaggca ctcccagcga aatcagtttc cccaatgacg
     5761 atttcgacaa agatccctat tgcagtagtc cgggttctga ctacaaagac gatgacgaca
     5821 agtgaagatc tgacaaaact cacacatgcc caccgtgccc agcacctgaa ctcctggggg
     5881 gaccgtcagt cttcctcttc cccccaaaac ccaaggacac cctcatgatc tcccggaccc
     5941 ctgaggtcac atgcgtggtg gtggacgtga gccacgaaga ccctgaggtc aagttcaact
     6001 ggtacgtgga cggcgtggag gtgcataatg ccaagacaaa gccgcgggag gagcagtaca
     6061 acagcacgta ccgtgtggtc agcgtcctca ccgtcctgca ccaggactgg ctgaatggca
     6121 aggagtacaa gtgcaaggtc tccaacaaag ccctcccagc ccccatcgag aaaaccatct
     6181 ccaaagccaa agggcagccc cgagaaccac aggtgtacac cctgccccca tcccgggagg
     6241 agatgaccaa gaaccaggtc agcctgacct gcctggtcaa aggcttctat cccagcgaca
     6301 tcgccgtgga gtgggagagc aatgggcagc cggagaacaa ctacaagacc acgcctcccg
     6361 tgctggactc cgacggctcc ttcttcctct acagcaagct caccgtggac aagagcaggt
     6421 ggcagcaggg gaacgtcttc t
//
