LOCUS       sequence-430305-        6591 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 6591)
  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 6591)
  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-ADGRV1 CTFdeltaTA-FLAG
FEATURES             Location/Qualifiers
     source          1..6591
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(547..570)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(2929..2955)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     sig_peptide     complement(2956..3015)
                     /codon_start=1
                     /product="human interleukin-2 signal sequence"
                     /label=IL-2 signal sequence
                     /translation="MYRMQLLSCIALSLALVTNS"
     LTR             3065..3333
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     promoter        complement(3346..3557)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     misc_feature    3594..3685
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     complement(3626..3645)
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     polyA_signal    3696..3744
                     /note="synthetic polyadenylation signal"
     rep_origin      3852..4440
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4341..4360
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        4520..4823
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             5416..5790
                     /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    5804..6198
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     polyA_signal    6302..6423
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6339..6358)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6393..6412
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 gtccagcacg ggaggcgtgg tcttgtagtt gttctccggc tgcccattgc tctcccactc
       61 cacggcgatg tcgctgggat agaagccttt gaccaggcag gtcaggctga cctggttctt
      121 ggtcatctcc tcccgggatg ggggcagggt gtacacctgt ggttctcggg gctgcccttt
      181 ggctttggag atggttttct cgatgggggc tgggagggct ttgttggaga ccttgcactt
      241 gtactccttg ccattcagcc agtcctggtg caggacggtg aggacgctga ccacacggta
      301 cgtgctgttg tactgctcct cccgcggctt tgtcttggca ttatgcacct ccacgccgtc
      361 cacgtaccag ttgaacttga cctcagggtc ttcgtggctc acgtccacca ccacgcatgt
      421 gacctcaggg gtccgggaga tcatgagggt gtccttgggt tttgggggga agaggaagac
      481 tgacggtccc cccaggagtt caggtgctgg gcacggtggg catgtgtgag ttttgtcaga
      541 tcttcacttg tcgtcatcgt ctttgtagtc agaacccaga tgggtgtcgg caattggaat
      601 tctccgcagc tccacaatct gagagtcggt cagggtgcca ccctcctggc ttccttgacc
      661 tgattcattg tcagagacgg agagtccggc gcctgtcttc agcgcgaaga tcaagtcatc
      721 gaactcctga gactcttcat ccgcgatcag gctcccttgg ccatagcctc cagaactagg
      781 gaaagtagct ccgttctggg gtgagggctt cagatcaggg gaagcctggc tgccctgttg
      841 aaaggaagcc cgctcccaat ccgggggtac ctcttccatg gcaccgatca gattttgtgt
      901 agacttggag atttcgcctc cggccggtgg catcccgctg cccggagtga aaaaggctgt
      961 ggagggacca ggatgaccat tcatctccac ggtatagctg gccttcatcg ggcaacacat
     1021 ctggttatgc aggataaaat agaccatgaa cacgtacaag ccctgcaagc tgttaaaaat
     1081 gacgaacagc accagcatcc aaaagtgacg atacgccata tgcaggccac cccaaagcca
     1141 ggtcacgcta atcaatgcga acaagtacaa gatcaggggg atttccgcgg catttgtacg
     1201 tccgcggaag acatcgtcgt atgccttcca ctggggttta acctgatagg catggataaa
     1261 tacgacgaag actacgacga gacaagtcag tggcaccagc gcagcggtga agagggcagc
     1321 gtaaacgtta gggataaagc agagatcgcc gtgaatcagg ccatagatct gggacatgga
     1381 ttggtggtag atgcccttca ggatcacgat cagaaggatc acgacgaacg ccggcaggcc
     1441 ccaagacagg aggaaaaaca gcaaatacct ccgctcggtg tgctcgtcat tcatgaccaa
     1501 gacgtaccag aagttcacac tctggatcag catccaggag aactggcaca ggtagaggta
     1561 gtgggtgacg gcagccatag cggaacagga ttcctcagcc agctggggtg aggcataagc
     1621 gctggcgagg aacagaatct gggtgcccag gctggcagcc atcatgtggg tcagcaactt
     1681 agcggcaaac atgctgtatc tcgcacagaa gatgtgggac agcactgcca ggcagagacc
     1741 gctgatacag atgaatccgc ttgtgaaaaa ggcttcattg tagctagaga gattgtcagt
     1801 acgttggaag taaaggttct cgttggtctg cgcatctttc agcgcttcat ccacggtctg
     1861 gcggccgctc gccgcgttaa tcaccgcggt gcgcaccgca taccaaaacg cgctcatctg
     1921 cggaatgttc ggcataattt cgcctttctg cgcgttttcc atggtcgccg caatgcgcgg
     1981 atctttcgcc agttcttctt catagctttt cagcgccacc gcgcccagcg gtttatcttt
     2041 gttcaccgct tccaggcctt catcggtcag cagatagttt tccagaaatt ctttcgccag
     2101 ttctttgttc gggctcgccg cgttaatgcc cgcgctcagc acgcccacaa acggtttgct
     2161 cggctggcct ttaaaggtcg gcagcacggt cacgccatag ttcactttgc tggtatcaat
     2221 gttgctccac gcccacgggc cgttaatggt catcgcggtt tcgcctttgt taaacgccgc
     2281 ttccgcaatg ctataatcgg tatccgcgtt catatgtttg tttttaatca gatccaccag
     2341 aaaggtcagg cccgctttcg cgcccgcgtt atccacgccc acatctttaa tatcatattt
     2401 gccgttttca tatttaaacg catagccgcc atccgccgca atcagcggcc aggtaaaata
     2461 cggttcctgc aggttaaaca tcagcgcgct tttgcctttc gctttcagtt ctttatccag
     2521 cgccggaatt tcttcccagg ttttcggcgg gttcggcagc agatctttgt tataaatcag
     2581 gctcagcgct tccaccgcaa tcggatacgc aatcagtttg ccgttatagc gcaccgcatc
     2641 ccaggtaaac ggatacagtt tatcctgaaa cgctttatcc ggggtaattt ccgccagcag
     2701 gccgctctgc gcatagccgc caaagcgatc atgcgcccaa aaaataatat ccgggccatc
     2761 gccggtcgcc gccacctgcg gaaatttttc ttccagttta tccggatgtt ccacggtcac
     2821 tttaatgccg gtatcttttt caaatttttt gcccacttcc gccaggccgt tatagccttt
     2881 atcgccgtta atccaaatca ccagtttgcc ttcttcaatt ttggatccag cgtaatctgg
     2941 aacatcgtat gggtacgaat tcgtgacaag tgcaagactt agtgcaatgc aagacaggag
     3001 ttgcatcctg tacatggtgg ccctccttcg ccggtgatct caggtaggcg ccggtcacag
     3061 cttggatctg taacggcgca gaacagaaaa cgaaacaaag acgtagagtt gagcaagcag
     3121 ggtcaggcaa agcgtggaga gccggctgag tctaggtagg ctccaaggga gcgccggaca
     3181 aaggcccggt ctcgacctga gctttaaact tacctagacg gcggacgcag ttcaggaggc
     3241 accacaggcg ggaggcggca gaacgcgact caaccggcgt ggatggcggc ctcaggtagg
     3301 gcggcgggcg cgtgaaggag agatgcgagc ccctcgaagc ttcagctgtg ttctggcggc
     3361 aaacccgttg cgaaaaagaa cgttcacggc gactactgca cttatatacg gttctccccc
     3421 accctcggga aaaaggcgga gccagtacac gacatcactt tcccagttta ccccgcgcca
     3481 ccttctctag gcacccgttc aattgccgac ccctcccccc aacttctcgg ggactgtggg
     3541 cgatgtgcgc tctgcccact gacgggcacc ggagcgatcg cagatccttc gatagagaaa
     3601 tgttctggca cctgcacttg cactggggac agcctatttt gctagtttgt tttgtttcgt
     3661 tttgttttga tggagagcgt atgttagtta cgattcacac aaaaaaccaa cacacagatg
     3721 taatgaaaat aaagatattt tattgcggcc gctgatttaa atgttaatta actagccatg
     3781 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     3841 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     3901 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     3961 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     4021 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     4081 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     4141 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     4201 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     4261 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     4321 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     4381 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     4441 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     4501 ttcttaatta acctgcaggc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
     4561 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
     4621 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
     4681 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
     4741 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
     4801 tattagtcat cgctattacc atgatgatgc ggttttggca gtacatcaat gggcgtggat
     4861 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
     4921 tttgactagt cagggcccca acccccccaa gcccccattt cacaacacgc tggcgctaca
     4981 ggcgcgtgac ttccccttgc tttggggcgg ggggctgaga ctcctatgtg ctccggattg
     5041 gtcaggcacg gccttcggcc ccgcctcctg ccaccgcaga ttggccgcta ggcctccccg
     5101 agcgccctgc ctccgagggc cggcgcacca taaaagaagc cgccctagcc acgtcccctc
     5161 gcagttcggc ggtcccgcgg gtctgtctca agcttctgcc ttctccctcc tgtgagtttg
     5221 gtaagtcact gactgtctat gcctgggaaa gggtgggcag gagatggggc agtgcaggaa
     5281 aagtggcact atgaaccctg cagccctagt ttgacaatta atcattggca tagtatatct
     5341 gcatagtata atacaactca ctatagcaat tgtactaacc ttcttctctt tcctctcctg
     5401 acaggaggag ccatcatggc caagttgacc agtgccgttc cggtgctcac cgcgcgcgac
     5461 gtcgccggag cggtcgagtt ctggaccgac cggctcgggt tctcccggga cttcgtggag
     5521 gacgacttcg ccggtgtggt ccgggacgac gtgaccctgt tcatcagcgc ggtccaggac
     5581 caggtggtgc cggacaacac cctggcctgg gtgtgggtgc gcggcctgga cgagctgtac
     5641 gccgagtggt cggaggtcgt gtccacgaac ttccgggacg cctccgggcc ggccatgacc
     5701 gagatcggcg agcagccgtg ggggcgggag ttcgccctgc gcgacccggc cggcaactgc
     5761 gtgcactttg tggcagagga gcaggactga ggataagcta gaagctcgct ttcttgctgt
     5821 ccaatttcta ttaaaggttc ctttgttccc taagtccaac tactaaactg ggggatatta
     5881 tgaagggcct tgagcatctg gattctgcct aataaaaaac atttattttc attgcaatga
     5941 tgtatttaaa ttatttctga atattttact aaaaagggaa tgtgggaggt cagtgcattt
     6001 aaaacataaa gaaatgaaga gctagttcaa accttgggaa aatacactat atcttaaact
     6061 ccatgaaaga aggtgaggct gcaaacagct aatgcacatt ggcaacagcc cctgatgcct
     6121 atgccttatt catccctcag aaaaggattc aagtagaggc ttgatttgga ggttaaagtt
     6181 ttgctatgct gtattttaga attaattcca taccacattt gtagaggttt tacttgcttt
     6241 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
     6301 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     6361 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     6421 ttatcatgtc tggccagcta gcactcattt acccggagac agggagaggc tcttctgcgt
     6481 gtagtggttg tgcagagcct cgtgcatcac ggagcatgag aagacgttcc cctgctgcca
     6541 cctgctcttg tccacggtga gcttgctgta gaggaagaag gagccgtcgg a
//
