LOCUS       sequence-430119-        6612 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 6612)
  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 6612)
  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 CTF-FLAG
FEATURES             Location/Qualifiers
     source          1..6612
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        49..260
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             273..541
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     sig_peptide     591..650
                     /codon_start=1
                     /product="human interleukin-2 signal sequence"
                     /label=IL-2 signal sequence
                     /translation="MYRMQLLSCIALSLALVTNS"
     CDS             651..677
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1785..1805
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             3057..3080
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    3795..3916
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3806..3825)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3860..3879
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     polyA_signal    4020..4414
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     CDS             complement(4428..4802)
                     /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        5395..5698
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(5778..6366)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5858..5877)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    6474..6522
                     /note="synthetic polyadenylation signal"
     misc_feature    join(6533..6612,1..12)
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     6573..6592
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 gaacatttct ctatcgaagg atctgcgatc gctccggtgc ccgtcagtgg gcagagcgca
       61 catcgcccac agtccccgag aagttggggg gaggggtcgg caattgaacg ggtgcctaga
      121 gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg
      181 agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg
      241 ggtttgccgc cagaacacag ctgaagcttc gaggggctcg catctctcct tcacgcgccc
      301 gccgccctac ctgaggccgc catccacgcc ggttgagtcg cgttctgccg cctcccgcct
      361 gtggtgcctc ctgaactgcg tccgccgtct aggtaagttt aaagctcagg tcgagaccgg
      421 gcctttgtcc ggcgctccct tggagcctac ctagactcag ccggctctcc acgctttgcc
      481 tgaccctgct tgctcaactc tacgtctttg tttcgttttc tgttctgcgc cgttacagat
      541 ccaagctgtg accggcgcct acctgagatc accggcgaag gagggccacc atgtacagga
      601 tgcaactcct gtcttgcatt gcactaagtc ttgcacttgt cacgaattcg tacccatacg
      661 atgttccaga ttacgctgga tccaaaattg aagaaggcaa actggtgatt tggattaacg
      721 gcgataaagg ctataacggc ctggcggaag tgggcaaaaa atttgaaaaa gataccggca
      781 ttaaagtgac cgtggaacat ccggataaac tggaagaaaa atttccgcag gtggcggcga
      841 ccggcgatgg cccggatatt attttttggg cgcatgatcg ctttggcggc tatgcgcaga
      901 gcggcctgct ggcggaaatt accccggata aagcgtttca ggataaactg tatccgttta
      961 cctgggatgc ggtgcgctat aacggcaaac tgattgcgta tccgattgcg gtggaagcgc
     1021 tgagcctgat ttataacaaa gatctgctgc cgaacccgcc gaaaacctgg gaagaaattc
     1081 cggcgctgga taaagaactg aaagcgaaag gcaaaagcgc gctgatgttt aacctgcagg
     1141 aaccgtattt tacctggccg ctgattgcgg cggatggcgg ctatgcgttt aaatatgaaa
     1201 acggcaaata tgatattaaa gatgtgggcg tggataacgc gggcgcgaaa gcgggcctga
     1261 cctttctggt ggatctgatt aaaaacaaac atatgaacgc ggataccgat tatagcattg
     1321 cggaagcggc gtttaacaaa ggcgaaaccg cgatgaccat taacggcccg tgggcgtgga
     1381 gcaacattga taccagcaaa gtgaactatg gcgtgaccgt gctgccgacc tttaaaggcc
     1441 agccgagcaa accgtttgtg ggcgtgctga gcgcgggcat taacgcggcg agcccgaaca
     1501 aagaactggc gaaagaattt ctggaaaact atctgctgac cgatgaaggc ctggaagcgg
     1561 tgaacaaaga taaaccgctg ggcgcggtgg cgctgaaaag ctatgaagaa gaactggcga
     1621 aagatccgcg cattgcggcg accatggaaa acgcgcagaa aggcgaaatt atgccgaaca
     1681 ttccgcagat gagcgcgttt tggtatgcgg tgcgcaccgc ggtgattaac gcggcgagcg
     1741 gccgccagac cgtggatgaa gcgctgaaag atgcgcagac caacgagaac ctttacttcc
     1801 aaagcgtcta cgcagtctac gcgcgtactg acaatctctc tagctacaat gaagcctttt
     1861 tcacaagcgg attcatctgt atcagcggtc tctgcctggc agtgctgtcc cacatcttct
     1921 gtgcgagata cagcatgttt gccgctaagt tgctgaccca catgatggct gccagcctgg
     1981 gcacccagat tctgttcctc gccagcgctt atgcctcacc ccagctggct gaggaatcct
     2041 gttccgctat ggctgccgtc acccactacc tctacctgtg ccagttctcc tggatgctga
     2101 tccagagtgt gaacttctgg tacgtcttgg tcatgaatga cgagcacacc gagcggaggt
     2161 atttgctgtt tttcctcctg tcttggggcc tgccggcgtt cgtcgtgatc cttctgatcg
     2221 tgatcctgaa gggcatctac caccaatcca tgtcccagat ctatggcctg attcacggcg
     2281 atctctgctt tatccctaac gtttacgctg ccctcttcac cgctgcgctg gtgccactga
     2341 cttgtctcgt cgtagtcttc gtcgtattta tccatgccta tcaggttaaa ccccagtgga
     2401 aggcatacga cgatgtcttc cgcggacgta caaatgccgc ggaaatcccc ctgatcttgt
     2461 acttgttcgc attgattagc gtgacctggc tttggggtgg cctgcatatg gcgtatcgtc
     2521 acttttggat gctggtgctg ttcgtcattt ttaacagctt gcagggcttg tacgtgttca
     2581 tggtctattt tatcctgcat aaccagatgt gttgcccgat gaaggccagc tataccgtgg
     2641 agatgaatgg tcatcctggt ccctccacag cctttttcac tccgggcagc gggatgccac
     2701 cggccggagg cgaaatctcc aagtctacac aaaatctgat cggtgccatg gaagaggtac
     2761 ccccggattg ggagcgggct tcctttcaac agggcagcca ggcttcccct gatctgaagc
     2821 cctcacccca gaacggagct actttcccta gttctggagg ctatggccaa gggagcctga
     2881 tcgcggatga agagtctcag gagttcgatg acttgatctt cgcgctgaag acaggcgccg
     2941 gactctccgt ctctgacaat gaatcaggtc aaggaagcca ggagggtggc accctgaccg
     3001 actctcagat tgtggagctg cggagaattc caattgccga cacccatctg ggttctgact
     3061 acaaagacga tgacgacaag tgaagatctg acaaaactca cacatgccca ccgtgcccag
     3121 cacctgaact cctgggggga ccgtcagtct tcctcttccc cccaaaaccc aaggacaccc
     3181 tcatgatctc ccggacccct gaggtcacat gcgtggtggt ggacgtgagc cacgaagacc
     3241 ctgaggtcaa gttcaactgg tacgtggacg gcgtggaggt gcataatgcc aagacaaagc
     3301 cgcgggagga gcagtacaac agcacgtacc gtgtggtcag cgtcctcacc gtcctgcacc
     3361 aggactggct gaatggcaag gagtacaagt gcaaggtctc caacaaagcc ctcccagccc
     3421 ccatcgagaa aaccatctcc aaagccaaag ggcagccccg agaaccacag gtgtacaccc
     3481 tgcccccatc ccgggaggag atgaccaaga accaggtcag cctgacctgc ctggtcaaag
     3541 gcttctatcc cagcgacatc gccgtggagt gggagagcaa tgggcagccg gagaacaact
     3601 acaagaccac gcctcccgtg ctggactccg acggctcctt cttcctctac agcaagctca
     3661 ccgtggacaa gagcaggtgg cagcagggga acgtcttctc atgctccgtg atgcacgagg
     3721 ctctgcacaa ccactacacg cagaagagcc tctccctgtc tccgggtaaa tgagtgctag
     3781 ctggccagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     3841 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     3901 ctgcaataaa caagttaaca acaaaaattg cattcatttt atgtttcagg ttcaggggga
     3961 ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtatgg aattaattct
     4021 aaaatacagc atagcaaaac tttaacctcc aaatcaagcc tctacttgaa tccttttctg
     4081 agggatgaat aaggcatagg catcaggggc tgttgccaat gtgcattagc tgtttgcagc
     4141 ctcaccttct ttcatggagt ttaagatata gtgtattttc ccaaggtttg aactagctct
     4201 tcatttcttt atgttttaaa tgcactgacc tcccacattc cctttttagt aaaatattca
     4261 gaaataattt aaatacatca ttgcaatgaa aataaatgtt ttttattagg cagaatccag
     4321 atgctcaagg cccttcataa tatcccccag tttagtagtt ggacttaggg aacaaaggaa
     4381 cctttaatag aaattggaca gcaagaaagc gagcttctag cttatcctca gtcctgctcc
     4441 tctgccacaa agtgcacgca gttgccggcc gggtcgcgca gggcgaactc ccgcccccac
     4501 ggctgctcgc cgatctcggt catggccggc ccggaggcgt cccggaagtt cgtggacacg
     4561 acctccgacc actcggcgta cagctcgtcc aggccgcgca cccacaccca ggccagggtg
     4621 ttgtccggca ccacctggtc ctggaccgcg ctgatgaaca gggtcacgtc gtcccggacc
     4681 acaccggcga agtcgtcctc cacgaagtcc cgggagaacc cgagccggtc ggtccagaac
     4741 tcgaccgctc cggcgacgtc gcgcgcggtg agcaccggaa cggcactggt caacttggcc
     4801 atgatggctc ctcctgtcag gagaggaaag agaagaaggt tagtacaatt gctatagtga
     4861 gttgtattat actatgcaga tatactatgc caatgattaa ttgtcaaact agggctgcag
     4921 ggttcatagt gccacttttc ctgcactgcc ccatctcctg cccacccttt cccaggcata
     4981 gacagtcagt gacttaccaa actcacagga gggagaaggc agaagcttga gacagacccg
     5041 cgggaccgcc gaactgcgag gggacgtggc tagggcggct tcttttatgg tgcgccggcc
     5101 ctcggaggca gggcgctcgg ggaggcctag cggccaatct gcggtggcag gaggcggggc
     5161 cgaaggccgt gcctgaccaa tccggagcac ataggagtct cagccccccg ccccaaagca
     5221 aggggaagtc acgcgcctgt agcgccagcg tgttgtgaaa tgggggcttg ggggggttgg
     5281 ggccctgact agtcaaaaca aactcccatt gacgtcaatg gggtggagac ttggaaatcc
     5341 ccgtgagtca aaccgctatc cacgcccatt gatgtactgc caaaaccgca tcatcatggt
     5401 aatagcgatg actaatacgt agatgtactg ccaagtagga aagtcccata aggtcatgta
     5461 ctgggcataa tgccaggcgg gccatttacc gtcattgacg tcaatagggg gcgtacttgg
     5521 catatgatac acttgatgta ctgccaagtg ggcagtttac cgtaaatact ccacccattg
     5581 acgtcaatgg aaagtcccta ttggcgttac tatgggaaca tacgtcatta ttgacgtcaa
     5641 tgggcggggg tcgttgggcg gtcagccagg cgggccattt accgtaagtt atgtaacgcc
     5701 tgcaggttaa ttaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     5761 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     5821 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     5881 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     5941 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     6001 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     6061 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     6121 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     6181 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     6241 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     6301 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     6361 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     6421 cgttaaggga ttttggtcat ggctagttaa ttaacattta aatcagcggc cgcaataaaa
     6481 tatctttatt ttcattacat ctgtgtgttg gttttttgtg tgaatcgtaa ctaacatacg
     6541 ctctccatca aaacaaaacg aaacaaaaca aactagcaaa ataggctgtc cccagtgcaa
     6601 gtgcaggtgc ca
//
