LOCUS       sequence-430284-        6201 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 6201)
  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 6201)
  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-ADGRF2 CTF-FLAG
FEATURES             Location/Qualifiers
     source          1..6201
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     41..60
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     polyA_signal    201..595
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     CDS             complement(609..983)
                     /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        1576..1879
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(1959..2547)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2039..2058)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    2655..2703
                     /note="synthetic polyadenylation signal"
     misc_feature    2714..2805
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     2754..2773
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        2842..3053
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             3066..3334
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     sig_peptide     3384..3443
                     /codon_start=1
                     /product="human interleukin-2 signal sequence"
                     /label=IL-2 signal sequence
                     /translation="MYRMQLLSCIALSLALVTNS"
     CDS             3444..3470
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             5439..5462
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    join(6177..6201,1..97)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(join(6188..6201,1..6))
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
       61 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
      121 tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa
      181 atgtggtatg gaattaattc taaaatacag catagcaaaa ctttaacctc caaatcaagc
      241 ctctacttga atccttttct gagggatgaa taaggcatag gcatcagggg ctgttgccaa
      301 tgtgcattag ctgtttgcag cctcaccttc tttcatggag tttaagatat agtgtatttt
      361 cccaaggttt gaactagctc ttcatttctt tatgttttaa atgcactgac ctcccacatt
      421 ccctttttag taaaatattc agaaataatt taaatacatc attgcaatga aaataaatgt
      481 tttttattag gcagaatcca gatgctcaag gcccttcata atatccccca gtttagtagt
      541 tggacttagg gaacaaagga acctttaata gaaattggac agcaagaaag cgagcttcta
      601 gcttatcctc agtcctgctc ctctgccaca aagtgcacgc agttgccggc cgggtcgcgc
      661 agggcgaact cccgccccca cggctgctcg ccgatctcgg tcatggccgg cccggaggcg
      721 tcccggaagt tcgtggacac gacctccgac cactcggcgt acagctcgtc caggccgcgc
      781 acccacaccc aggccagggt gttgtccggc accacctggt cctggaccgc gctgatgaac
      841 agggtcacgt cgtcccggac cacaccggcg aagtcgtcct ccacgaagtc ccgggagaac
      901 ccgagccggt cggtccagaa ctcgaccgct ccggcgacgt cgcgcgcggt gagcaccgga
      961 acggcactgg tcaacttggc catgatggct cctcctgtca ggagaggaaa gagaagaagg
     1021 ttagtacaat tgctatagtg agttgtatta tactatgcag atatactatg ccaatgatta
     1081 attgtcaaac tagggctgca gggttcatag tgccactttt cctgcactgc cccatctcct
     1141 gcccaccctt tcccaggcat agacagtcag tgacttacca aactcacagg agggagaagg
     1201 cagaagcttg agacagaccc gcgggaccgc cgaactgcga ggggacgtgg ctagggcggc
     1261 ttcttttatg gtgcgccggc cctcggaggc agggcgctcg gggaggccta gcggccaatc
     1321 tgcggtggca ggaggcgggg ccgaaggccg tgcctgacca atccggagca cataggagtc
     1381 tcagcccccc gccccaaagc aaggggaagt cacgcgcctg tagcgccagc gtgttgtgaa
     1441 atgggggctt gggggggttg gggccctgac tagtcaaaac aaactcccat tgacgtcaat
     1501 ggggtggaga cttggaaatc cccgtgagtc aaaccgctat ccacgcccat tgatgtactg
     1561 ccaaaaccgc atcatcatgg taatagcgat gactaatacg tagatgtact gccaagtagg
     1621 aaagtcccat aaggtcatgt actgggcata atgccaggcg ggccatttac cgtcattgac
     1681 gtcaataggg ggcgtacttg gcatatgata cacttgatgt actgccaagt gggcagttta
     1741 ccgtaaatac tccacccatt gacgtcaatg gaaagtccct attggcgtta ctatgggaac
     1801 atacgtcatt attgacgtca atgggcgggg gtcgttgggc ggtcagccag gcgggccatt
     1861 taccgtaagt tatgtaacgc ctgcaggtta attaagaaca tgtgagcaaa aggccagcaa
     1921 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     1981 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     2041 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     2101 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     2161 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     2221 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     2281 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     2341 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     2401 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     2461 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     2521 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     2581 gctcagtgga acgaaaactc acgttaaggg attttggtca tggctagtta attaacattt
     2641 aaatcagcgg ccgcaataaa atatctttat tttcattaca tctgtgtgtt ggttttttgt
     2701 gtgaatcgta actaacatac gctctccatc aaaacaaaac gaaacaaaac aaactagcaa
     2761 aataggctgt ccccagtgca agtgcaggtg ccagaacatt tctctatcga aggatctgcg
     2821 atcgctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg
     2881 ggggaggggt cggcaattga acgggtgcct agagaaggtg gcgcggggta aactgggaaa
     2941 gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg
     3001 cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca cagctgaagc
     3061 ttcgaggggc tcgcatctct ccttcacgcg cccgccgccc tacctgaggc cgccatccac
     3121 gccggttgag tcgcgttctg ccgcctcccg cctgtggtgc ctcctgaact gcgtccgccg
     3181 tctaggtaag tttaaagctc aggtcgagac cgggcctttg tccggcgctc ccttggagcc
     3241 tacctagact cagccggctc tccacgcttt gcctgaccct gcttgctcaa ctctacgtct
     3301 ttgtttcgtt ttctgttctg cgccgttaca gatccaagct gtgaccggcg cctacctgag
     3361 atcaccggcg aaggagggcc accatgtaca ggatgcaact cctgtcttgc attgcactaa
     3421 gtcttgcact tgtcacgaat tcgtacccat acgatgttcc agattacgct ggatccaaaa
     3481 ttgaagaagg caaactggtg atttggatta acggcgataa aggctataac ggcctggcgg
     3541 aagtgggcaa aaaatttgaa aaagataccg gcattaaagt gaccgtggaa catccggata
     3601 aactggaaga aaaatttccg caggtggcgg cgaccggcga tggcccggat attatttttt
     3661 gggcgcatga tcgctttggc ggctatgcgc agagcggcct gctggcggaa attaccccgg
     3721 ataaagcgtt tcaggataaa ctgtatccgt ttacctggga tgcggtgcgc tataacggca
     3781 aactgattgc gtatccgatt gcggtggaag cgctgagcct gatttataac aaagatctgc
     3841 tgccgaaccc gccgaaaacc tgggaagaaa ttccggcgct ggataaagaa ctgaaagcga
     3901 aaggcaaaag cgcgctgatg tttaacctgc aggaaccgta ttttacctgg ccgctgattg
     3961 cggcggatgg cggctatgcg tttaaatatg aaaacggcaa atatgatatt aaagatgtgg
     4021 gcgtggataa cgcgggcgcg aaagcgggcc tgacctttct ggtggatctg attaaaaaca
     4081 aacatatgaa cgcggatacc gattatagca ttgcggaagc ggcgtttaac aaaggcgaaa
     4141 ccgcgatgac cattaacggc ccgtgggcgt ggagcaacat tgataccagc aaagtgaact
     4201 atggcgtgac cgtgctgccg acctttaaag gccagccgag caaaccgttt gtgggcgtgc
     4261 tgagcgcggg cattaacgcg gcgagcccga acaaagaact ggcgaaagaa tttctggaaa
     4321 actatctgct gaccgatgaa ggcctggaag cggtgaacaa agataaaccg ctgggcgcgg
     4381 tggcgctgaa aagctatgaa gaagaactgg cgaaagatcc gcgcattgcg gcgaccatgg
     4441 aaaacgcgca gaaaggcgaa attatgccga acattccgca gatgagcgcg ttttggtatg
     4501 cggtgcgcac cgcggtgatt aacgcggcga gcggccgcca gaccgtggat gaagcgctga
     4561 aagatgcgca gaccaacgag aacctttact tccaaacaag cttctctatc ctgatgtccc
     4621 cgcacatcct ggagagcctc atcctcacct acatcactta tgttggtctg ggcatttcta
     4681 tctgttcact gatcttgtgt cttagcatcg aggtgctggt atggagccaa gtaacaaaga
     4741 ctgagatcac atacctgcgt cacgtgtgta tcgtgaacat cgctgcgacc ctgcttatgg
     4801 cggacgtgtg gtttatcgtt gcctcttttc tgagcggacc tattacgcat cacaagggat
     4861 gtgtcgcggc tacctttttc gttcatttct tttacctgag cgtgtttttc tggatgctgg
     4921 ctaaggcttt gctcatcctg tatggcatta tgatcgtgtt ccacactctg cccaagtccg
     4981 tgctggtcgc gagccttttc tctgtcggct atggctgtcc cctggccatt gctgccatta
     5041 ctgtggcagc gacagagccc ggaaagggct atctgcgccc ggagatctgc tggctgaact
     5101 gggatatgac gaaggctctg ctcgcgttcg tcatcccggc tttggccatc gtggtcgtaa
     5161 acctgattac agtgaccctg gtgatcgtga agacccagcg tgctgccatc ggtaacagca
     5221 tgtttcagga ggtgagggcg attgttcgga tttctaagaa cattgccatc ctgacacccc
     5281 tgctcggcct gacgtgggga ttcggcgttg caaccgtaat cgatgaccgc tccctggcct
     5341 tccatatcat ttttagcctc ctgaatgcct tccaagtgtc ccctgacgcg agtgatcagg
     5401 tgcagtcaga gcgcatccat gaagatgtgc tgggttctga ctacaaagac gatgacgaca
     5461 agtgaagatc tgacaaaact cacacatgcc caccgtgccc agcacctgaa ctcctggggg
     5521 gaccgtcagt cttcctcttc cccccaaaac ccaaggacac cctcatgatc tcccggaccc
     5581 ctgaggtcac atgcgtggtg gtggacgtga gccacgaaga ccctgaggtc aagttcaact
     5641 ggtacgtgga cggcgtggag gtgcataatg ccaagacaaa gccgcgggag gagcagtaca
     5701 acagcacgta ccgtgtggtc agcgtcctca ccgtcctgca ccaggactgg ctgaatggca
     5761 aggagtacaa gtgcaaggtc tccaacaaag ccctcccagc ccccatcgag aaaaccatct
     5821 ccaaagccaa agggcagccc cgagaaccac aggtgtacac cctgccccca tcccgggagg
     5881 agatgaccaa gaaccaggtc agcctgacct gcctggtcaa aggcttctat cccagcgaca
     5941 tcgccgtgga gtgggagagc aatgggcagc cggagaacaa ctacaagacc acgcctcccg
     6001 tgctggactc cgacggctcc ttcttcctct acagcaagct caccgtggac aagagcaggt
     6061 ggcagcaggg gaacgtcttc tcatgctccg tgatgcacga ggctctgcac aaccactaca
     6121 cgcagaagag cctctccctg tctccgggta aatgagtgct agctggccag acatgataag
     6181 atacattgat gagtttggac a
//
