LOCUS       sequence-430302-        7248 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 7248)
  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 7248)
  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-ADGRL2 CTFdeltaTA-FLAG
FEATURES             Location/Qualifiers
     source          1..7248
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(90..113)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(2001..2021)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQA"
     CDS             complement(3129..3155)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     sig_peptide     complement(3156..3215)
                     /codon_start=1
                     /product="human interleukin-2 signal sequence"
                     /label=IL-2 signal sequence
                     /translation="MYRMQLLSCIALSLALVTNS"
     LTR             3265..3533
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     promoter        complement(3546..3757)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     misc_feature    3794..3885
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     complement(3826..3845)
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     polyA_signal    3896..3944
                     /note="synthetic polyadenylation signal"
     rep_origin      4052..4640
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4541..4560
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        4720..5023
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             5616..5990
                     /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    6004..6398
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     polyA_signal    6502..6623
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6539..6558)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6593..6612
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 ggttttgggg ggaagaggaa gactgacggt ccccccagga gttcaggtgc tgggcacggt
       61 gggcatgtgt gagttttgtc agatcttcac ttgtcgtcat cgtctttgta gtcagaaccc
      121 agagaggtga ccagctgcat ttgtccttcg cgcacgtcgc cttctgggat acatccttct
      181 ttgttgatag ggataatgta gccgtcagaa tttccccgag agatctgata gcacatttga
      241 agctgatgac cggctcccag attcggcatg ctcttatagt agatatcttc gttctcagag
      301 cgacgggacg ggctcagatc ctcttccata tcgggggaag attcgggata gggtgagtca
      361 cgcaggttgg gcatggatgt gtagaggctg tcgcgattgg ggctctgcag atggtcctcg
      421 gcctcggcgg tcagctgtga cacatagcta tccgtaccct cactttttac ctttttctgg
      481 ggctggtaca ggagagaatg ggttctctgc gggatcaggg gagcttcgag ctccttgtga
      541 tggagctcca gcccagggtt gtcggaatgc atcagggatg aggcgtcggc cacgatagcg
      601 tcatcctctg agctggagcc accaataacg ggtttcacgg gcagagtcag ctccaggtta
      661 tgggtcttgg agctgcctct caggttattg tggaccagct cagagataat catcttctcg
      721 aaggctgtgt cgttcagaga caggccgcaa tcaacgacct gcacggagtc gttgtaatca
      781 cctttgtgca gtgagtatga gttattaaag ttgccgttca gtggcagggt gtccatagca
      841 ctcgtgtctc ttgcgttatt caggctgtga cccggagaat tgaagacagg ggcactgggt
      901 gcgttgcata ccacggtctc ggccagaagc gtattgtatg ggttattagt cccgtgggga
      961 cgcagaagag ggtttgtaag cagatagttg ccggtcatgc cctggttcag ggttgacgtg
     1021 gagttaatgt ctccggagat gaagctagac tcggactgtt ttctcacggt atcgttccac
     1081 atgcgcctga ttcttgactg ggtaccgcta gagtagcggg cggaggttct agtggtagaa
     1141 gcttttactg agctgtgtgg agactctgtg ggcaggcctc cacagcaata ggaatgccta
     1201 aaacacttac cgtactcttt acgaaccttt ttctgcaatg cacagtggaa gataaagata
     1261 aagacgccct ggaaagcgtt gaagatggtg aaaaggtagg ccataacgat ggtctcttcg
     1321 ttaatgaaga gcaagccgaa ggaccaggtc agccccagaa gacacaacag ggcgaatgcg
     1381 cccagaaccc aactcttgat gttctccagg cgagatgagt caggcttcag tgtgttgctg
     1441 tgcttgacca ttttacagag agtgatgaca aggaagataa tgttcagaag gataatgaac
     1501 gtcacggggc cgatgaatga ccagatgaaa tagttgtcga cgtgaagcca gcaggccttc
     1561 tccgtaccgt aacttttata gtcgatagct gcagatacgc ccacgacggt ggcagggaac
     1621 aggtaaccag caacatagta atactttttg cgagaatact cggattcaaa cacctccacc
     1681 agcatcaggt acagctgaac gccttccagg cacatccaag caaaggctgc cagaaagaaa
     1741 aagtggagca gaccagcaaa gatcggacag gcgatggcgt atttcgtttt gtcgatgccg
     1801 atcaagaaga tgaactcggc aatgaacagg ttgatgcaca agttcttgtg gatagtattc
     1861 cggtcagact gaagacccct gaaaaaacaa aaagtaaaaa tgcagatggc gaggcaaacc
     1921 aaactgatca caatacccac ccaggtgatg acggtgagca aaagctcgtg cacaccatcc
     1981 ttgtaagcaa tctcccggtg ggcttggaag taaaggttct cgttggtctg cgcatctttc
     2041 agcgcttcat ccacggtctg gcggccgctc gccgcgttaa tcaccgcggt gcgcaccgca
     2101 taccaaaacg cgctcatctg cggaatgttc ggcataattt cgcctttctg cgcgttttcc
     2161 atggtcgccg caatgcgcgg atctttcgcc agttcttctt catagctttt cagcgccacc
     2221 gcgcccagcg gtttatcttt gttcaccgct tccaggcctt catcggtcag cagatagttt
     2281 tccagaaatt ctttcgccag ttctttgttc gggctcgccg cgttaatgcc cgcgctcagc
     2341 acgcccacaa acggtttgct cggctggcct ttaaaggtcg gcagcacggt cacgccatag
     2401 ttcactttgc tggtatcaat gttgctccac gcccacgggc cgttaatggt catcgcggtt
     2461 tcgcctttgt taaacgccgc ttccgcaatg ctataatcgg tatccgcgtt catatgtttg
     2521 tttttaatca gatccaccag aaaggtcagg cccgctttcg cgcccgcgtt atccacgccc
     2581 acatctttaa tatcatattt gccgttttca tatttaaacg catagccgcc atccgccgca
     2641 atcagcggcc aggtaaaata cggttcctgc aggttaaaca tcagcgcgct tttgcctttc
     2701 gctttcagtt ctttatccag cgccggaatt tcttcccagg ttttcggcgg gttcggcagc
     2761 agatctttgt tataaatcag gctcagcgct tccaccgcaa tcggatacgc aatcagtttg
     2821 ccgttatagc gcaccgcatc ccaggtaaac ggatacagtt tatcctgaaa cgctttatcc
     2881 ggggtaattt ccgccagcag gccgctctgc gcatagccgc caaagcgatc atgcgcccaa
     2941 aaaataatat ccgggccatc gccggtcgcc gccacctgcg gaaatttttc ttccagttta
     3001 tccggatgtt ccacggtcac tttaatgccg gtatcttttt caaatttttt gcccacttcc
     3061 gccaggccgt tatagccttt atcgccgtta atccaaatca ccagtttgcc ttcttcaatt
     3121 ttggatccag cgtaatctgg aacatcgtat gggtacgaat tcgtgacaag tgcaagactt
     3181 agtgcaatgc aagacaggag ttgcatcctg tacatggtgg ccctccttcg ccggtgatct
     3241 caggtaggcg ccggtcacag cttggatctg taacggcgca gaacagaaaa cgaaacaaag
     3301 acgtagagtt gagcaagcag ggtcaggcaa agcgtggaga gccggctgag tctaggtagg
     3361 ctccaaggga gcgccggaca aaggcccggt ctcgacctga gctttaaact tacctagacg
     3421 gcggacgcag ttcaggaggc accacaggcg ggaggcggca gaacgcgact caaccggcgt
     3481 ggatggcggc ctcaggtagg gcggcgggcg cgtgaaggag agatgcgagc ccctcgaagc
     3541 ttcagctgtg ttctggcggc aaacccgttg cgaaaaagaa cgttcacggc gactactgca
     3601 cttatatacg gttctccccc accctcggga aaaaggcgga gccagtacac gacatcactt
     3661 tcccagttta ccccgcgcca ccttctctag gcacccgttc aattgccgac ccctcccccc
     3721 aacttctcgg ggactgtggg cgatgtgcgc tctgcccact gacgggcacc ggagcgatcg
     3781 cagatccttc gatagagaaa tgttctggca cctgcacttg cactggggac agcctatttt
     3841 gctagtttgt tttgtttcgt tttgttttga tggagagcgt atgttagtta cgattcacac
     3901 aaaaaaccaa cacacagatg taatgaaaat aaagatattt tattgcggcc gctgatttaa
     3961 atgttaatta actagccatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     4021 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     4081 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     4141 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     4201 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     4261 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     4321 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     4381 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     4441 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     4501 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     4561 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     4621 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     4681 gctggccttt tgctcacatg ttcttaatta acctgcaggc gttacataac ttacggtaaa
     4741 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
     4801 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
     4861 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
     4921 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
     4981 tacttggcag tacatctacg tattagtcat cgctattacc atgatgatgc ggttttggca
     5041 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
     5101 tgacgtcaat gggagtttgt tttgactagt cagggcccca acccccccaa gcccccattt
     5161 cacaacacgc tggcgctaca ggcgcgtgac ttccccttgc tttggggcgg ggggctgaga
     5221 ctcctatgtg ctccggattg gtcaggcacg gccttcggcc ccgcctcctg ccaccgcaga
     5281 ttggccgcta ggcctccccg agcgccctgc ctccgagggc cggcgcacca taaaagaagc
     5341 cgccctagcc acgtcccctc gcagttcggc ggtcccgcgg gtctgtctca agcttctgcc
     5401 ttctccctcc tgtgagtttg gtaagtcact gactgtctat gcctgggaaa gggtgggcag
     5461 gagatggggc agtgcaggaa aagtggcact atgaaccctg cagccctagt ttgacaatta
     5521 atcattggca tagtatatct gcatagtata atacaactca ctatagcaat tgtactaacc
     5581 ttcttctctt tcctctcctg acaggaggag ccatcatggc caagttgacc agtgccgttc
     5641 cggtgctcac cgcgcgcgac gtcgccggag cggtcgagtt ctggaccgac cggctcgggt
     5701 tctcccggga cttcgtggag gacgacttcg ccggtgtggt ccgggacgac gtgaccctgt
     5761 tcatcagcgc ggtccaggac caggtggtgc cggacaacac cctggcctgg gtgtgggtgc
     5821 gcggcctgga cgagctgtac gccgagtggt cggaggtcgt gtccacgaac ttccgggacg
     5881 cctccgggcc ggccatgacc gagatcggcg agcagccgtg ggggcgggag ttcgccctgc
     5941 gcgacccggc cggcaactgc gtgcactttg tggcagagga gcaggactga ggataagcta
     6001 gaagctcgct ttcttgctgt ccaatttcta ttaaaggttc ctttgttccc taagtccaac
     6061 tactaaactg ggggatatta tgaagggcct tgagcatctg gattctgcct aataaaaaac
     6121 atttattttc attgcaatga tgtatttaaa ttatttctga atattttact aaaaagggaa
     6181 tgtgggaggt cagtgcattt aaaacataaa gaaatgaaga gctagttcaa accttgggaa
     6241 aatacactat atcttaaact ccatgaaaga aggtgaggct gcaaacagct aatgcacatt
     6301 ggcaacagcc cctgatgcct atgccttatt catccctcag aaaaggattc aagtagaggc
     6361 ttgatttgga ggttaaagtt ttgctatgct gtattttaga attaattcca taccacattt
     6421 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa
     6481 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
     6541 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     6601 tccaaactca tcaatgtatc ttatcatgtc tggccagcta gcactcattt acccggagac
     6661 agggagaggc tcttctgcgt gtagtggttg tgcagagcct cgtgcatcac ggagcatgag
     6721 aagacgttcc cctgctgcca cctgctcttg tccacggtga gcttgctgta gaggaagaag
     6781 gagccgtcgg agtccagcac gggaggcgtg gtcttgtagt tgttctccgg ctgcccattg
     6841 ctctcccact ccacggcgat gtcgctggga tagaagcctt tgaccaggca ggtcaggctg
     6901 acctggttct tggtcatctc ctcccgggat gggggcaggg tgtacacctg tggttctcgg
     6961 ggctgccctt tggctttgga gatggttttc tcgatggggg ctgggagggc tttgttggag
     7021 accttgcact tgtactcctt gccattcagc cagtcctggt gcaggacggt gaggacgctg
     7081 accacacggt acgtgctgtt gtactgctcc tcccgcggct ttgtcttggc attatgcacc
     7141 tccacgccgt ccacgtacca gttgaacttg acctcagggt cttcgtggct cacgtccacc
     7201 accacgcatg tgacctcagg ggtccgggag atcatgaggg tgtccttg
//
