LOCUS       sequence-430000-        6180 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 6180)
  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 6180)
  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 CTFdeltaTA-FLAG
FEATURES             Location/Qualifiers
     source          1..6180
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        31..242
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             255..523
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     sig_peptide     573..632
                     /codon_start=1
                     /product="human interleukin-2 signal sequence"
                     /label=IL-2 signal sequence
                     /translation="MYRMQLLSCIALSLALVTNS"
     CDS             633..659
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1767..1787
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             2607..2630
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    3345..3466
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3356..3375)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3410..3429
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     polyA_signal    3570..3964
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     CDS             complement(3978..4352)
                     /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        4945..5248
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(5328..5916)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5408..5427)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    6024..6072
                     /note="synthetic polyadenylation signal"
     misc_feature    6083..6174
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     6123..6142
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggatctgcga tcgctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg
       61 agaagttggg gggaggggtc ggcaattgaa cgggtgccta gagaaggtgg cgcggggtaa
      121 actgggaaag tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt
      181 atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac
      241 agctgaagct tcgaggggct cgcatctctc cttcacgcgc ccgccgccct acctgaggcc
      301 gccatccacg ccggttgagt cgcgttctgc cgcctcccgc ctgtggtgcc tcctgaactg
      361 cgtccgccgt ctaggtaagt ttaaagctca ggtcgagacc gggcctttgt ccggcgctcc
      421 cttggagcct acctagactc agccggctct ccacgctttg cctgaccctg cttgctcaac
      481 tctacgtctt tgtttcgttt tctgttctgc gccgttacag atccaagctg tgaccggcgc
      541 ctacctgaga tcaccggcga aggagggcca ccatgtacag gatgcaactc ctgtcttgca
      601 ttgcactaag tcttgcactt gtcacgaatt cgtacccata cgatgttcca gattacgctg
      661 gatccaaaat tgaagaaggc aaactggtga tttggattaa cggcgataaa ggctataacg
      721 gcctggcgga agtgggcaaa aaatttgaaa aagataccgg cattaaagtg accgtggaac
      781 atccggataa actggaagaa aaatttccgc aggtggcggc gaccggcgat ggcccggata
      841 ttattttttg ggcgcatgat cgctttggcg gctatgcgca gagcggcctg ctggcggaaa
      901 ttaccccgga taaagcgttt caggataaac tgtatccgtt tacctgggat gcggtgcgct
      961 ataacggcaa actgattgcg tatccgattg cggtggaagc gctgagcctg atttataaca
     1021 aagatctgct gccgaacccg ccgaaaacct gggaagaaat tccggcgctg gataaagaac
     1081 tgaaagcgaa aggcaaaagc gcgctgatgt ttaacctgca ggaaccgtat tttacctggc
     1141 cgctgattgc ggcggatggc ggctatgcgt ttaaatatga aaacggcaaa tatgatatta
     1201 aagatgtggg cgtggataac gcgggcgcga aagcgggcct gacctttctg gtggatctga
     1261 ttaaaaacaa acatatgaac gcggataccg attatagcat tgcggaagcg gcgtttaaca
     1321 aaggcgaaac cgcgatgacc attaacggcc cgtgggcgtg gagcaacatt gataccagca
     1381 aagtgaacta tggcgtgacc gtgctgccga cctttaaagg ccagccgagc aaaccgtttg
     1441 tgggcgtgct gagcgcgggc attaacgcgg cgagcccgaa caaagaactg gcgaaagaat
     1501 ttctggaaaa ctatctgctg accgatgaag gcctggaagc ggtgaacaaa gataaaccgc
     1561 tgggcgcggt ggcgctgaaa agctatgaag aagaactggc gaaagatccg cgcattgcgg
     1621 cgaccatgga aaacgcgcag aaaggcgaaa ttatgccgaa cattccgcag atgagcgcgt
     1681 tttggtatgc ggtgcgcacc gcggtgatta acgcggcgag cggccgccag accgtggatg
     1741 aagcgctgaa agatgcgcag accaacgaga acctttactt ccaatccccg cacatcctgg
     1801 agagcctcat cctcacctac atcacttatg ttggtctggg catttctatc tgttcactga
     1861 tcttgtgtct tagcatcgag gtgctggtat ggagccaagt aacaaagact gagatcacat
     1921 acctgcgtca cgtgtgtatc gtgaacatcg ctgcgaccct gcttatggcg gacgtgtggt
     1981 ttatcgttgc ctcttttctg agcggaccta ttacgcatca caagggatgt gtcgcggcta
     2041 cctttttcgt tcatttcttt tacctgagcg tgtttttctg gatgctggct aaggctttgc
     2101 tcatcctgta tggcattatg atcgtgttcc acactctgcc caagtccgtg ctggtcgcga
     2161 gccttttctc tgtcggctat ggctgtcccc tggccattgc tgccattact gtggcagcga
     2221 cagagcccgg aaagggctat ctgcgcccgg agatctgctg gctgaactgg gatatgacga
     2281 aggctctgct cgcgttcgtc atcccggctt tggccatcgt ggtcgtaaac ctgattacag
     2341 tgaccctggt gatcgtgaag acccagcgtg ctgccatcgg taacagcatg tttcaggagg
     2401 tgagggcgat tgttcggatt tctaagaaca ttgccatcct gacacccctg ctcggcctga
     2461 cgtggggatt cggcgttgca accgtaatcg atgaccgctc cctggccttc catatcattt
     2521 ttagcctcct gaatgccttc caagtgtccc ctgacgcgag tgatcaggtg cagtcagagc
     2581 gcatccatga agatgtgctg ggttctgact acaaagacga tgacgacaag tgaagatctg
     2641 acaaaactca cacatgccca ccgtgcccag cacctgaact cctgggggga ccgtcagtct
     2701 tcctcttccc cccaaaaccc aaggacaccc tcatgatctc ccggacccct gaggtcacat
     2761 gcgtggtggt ggacgtgagc cacgaagacc ctgaggtcaa gttcaactgg tacgtggacg
     2821 gcgtggaggt gcataatgcc aagacaaagc cgcgggagga gcagtacaac agcacgtacc
     2881 gtgtggtcag cgtcctcacc gtcctgcacc aggactggct gaatggcaag gagtacaagt
     2941 gcaaggtctc caacaaagcc ctcccagccc ccatcgagaa aaccatctcc aaagccaaag
     3001 ggcagccccg agaaccacag gtgtacaccc tgcccccatc ccgggaggag atgaccaaga
     3061 accaggtcag cctgacctgc ctggtcaaag gcttctatcc cagcgacatc gccgtggagt
     3121 gggagagcaa tgggcagccg gagaacaact acaagaccac gcctcccgtg ctggactccg
     3181 acggctcctt cttcctctac agcaagctca ccgtggacaa gagcaggtgg cagcagggga
     3241 acgtcttctc atgctccgtg atgcacgagg ctctgcacaa ccactacacg cagaagagcc
     3301 tctccctgtc tccgggtaaa tgagtgctag ctggccagac atgataagat acattgatga
     3361 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     3421 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     3481 cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa
     3541 cctctacaaa tgtggtatgg aattaattct aaaatacagc atagcaaaac tttaacctcc
     3601 aaatcaagcc tctacttgaa tccttttctg agggatgaat aaggcatagg catcaggggc
     3661 tgttgccaat gtgcattagc tgtttgcagc ctcaccttct ttcatggagt ttaagatata
     3721 gtgtattttc ccaaggtttg aactagctct tcatttcttt atgttttaaa tgcactgacc
     3781 tcccacattc cctttttagt aaaatattca gaaataattt aaatacatca ttgcaatgaa
     3841 aataaatgtt ttttattagg cagaatccag atgctcaagg cccttcataa tatcccccag
     3901 tttagtagtt ggacttaggg aacaaaggaa cctttaatag aaattggaca gcaagaaagc
     3961 gagcttctag cttatcctca gtcctgctcc tctgccacaa agtgcacgca gttgccggcc
     4021 gggtcgcgca gggcgaactc ccgcccccac ggctgctcgc cgatctcggt catggccggc
     4081 ccggaggcgt cccggaagtt cgtggacacg acctccgacc actcggcgta cagctcgtcc
     4141 aggccgcgca cccacaccca ggccagggtg ttgtccggca ccacctggtc ctggaccgcg
     4201 ctgatgaaca gggtcacgtc gtcccggacc acaccggcga agtcgtcctc cacgaagtcc
     4261 cgggagaacc cgagccggtc ggtccagaac tcgaccgctc cggcgacgtc gcgcgcggtg
     4321 agcaccggaa cggcactggt caacttggcc atgatggctc ctcctgtcag gagaggaaag
     4381 agaagaaggt tagtacaatt gctatagtga gttgtattat actatgcaga tatactatgc
     4441 caatgattaa ttgtcaaact agggctgcag ggttcatagt gccacttttc ctgcactgcc
     4501 ccatctcctg cccacccttt cccaggcata gacagtcagt gacttaccaa actcacagga
     4561 gggagaaggc agaagcttga gacagacccg cgggaccgcc gaactgcgag gggacgtggc
     4621 tagggcggct tcttttatgg tgcgccggcc ctcggaggca gggcgctcgg ggaggcctag
     4681 cggccaatct gcggtggcag gaggcggggc cgaaggccgt gcctgaccaa tccggagcac
     4741 ataggagtct cagccccccg ccccaaagca aggggaagtc acgcgcctgt agcgccagcg
     4801 tgttgtgaaa tgggggcttg ggggggttgg ggccctgact agtcaaaaca aactcccatt
     4861 gacgtcaatg gggtggagac ttggaaatcc ccgtgagtca aaccgctatc cacgcccatt
     4921 gatgtactgc caaaaccgca tcatcatggt aatagcgatg actaatacgt agatgtactg
     4981 ccaagtagga aagtcccata aggtcatgta ctgggcataa tgccaggcgg gccatttacc
     5041 gtcattgacg tcaatagggg gcgtacttgg catatgatac acttgatgta ctgccaagtg
     5101 ggcagtttac cgtaaatact ccacccattg acgtcaatgg aaagtcccta ttggcgttac
     5161 tatgggaaca tacgtcatta ttgacgtcaa tgggcggggg tcgttgggcg gtcagccagg
     5221 cgggccattt accgtaagtt atgtaacgcc tgcaggttaa ttaagaacat gtgagcaaaa
     5281 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     5341 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     5401 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     5461 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     5521 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     5581 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     5641 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     5701 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     5761 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     5821 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     5881 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     5941 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat ggctagttaa
     6001 ttaacattta aatcagcggc cgcaataaaa tatctttatt ttcattacat ctgtgtgttg
     6061 gttttttgtg tgaatcgtaa ctaacatacg ctctccatca aaacaaaacg aaacaaaaca
     6121 aactagcaaa ataggctgtc cccagtgcaa gtgcaggtgc cagaacattt ctctatcgaa
//
