LOCUS       sequence-430276-        7752 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 7752)
  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 7752)
  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-ADGRC3 CTF-FLAG
FEATURES             Location/Qualifiers
     source          1..7752
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    165..256
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     complement(197..216)
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     polyA_signal    267..315
                     /note="synthetic polyadenylation signal"
     rep_origin      423..1011
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     912..931
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        1091..1394
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             1987..2361
                     /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    2375..2769
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     polyA_signal    2873..2994
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2910..2929)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2964..2983
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(3709..3732)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(7252..7278)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     sig_peptide     complement(7279..7338)
                     /codon_start=1
                     /product="human interleukin-2 signal sequence"
                     /label=IL-2 signal sequence
                     /translation="MYRMQLLSCIALSLALVTNS"
     LTR             7388..7656
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     promoter        complement(join(7669..7752,1..128))
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
ORIGIN
        1 aaaaaggcgg agccagtaca cgacatcact ttcccagttt accccgcgcc accttctcta
       61 ggcacccgtt caattgccga cccctccccc caacttctcg gggactgtgg gcgatgtgcg
      121 ctctgcccac tgacgggcac cggagcgatc gcagatcctt cgatagagaa atgttctggc
      181 acctgcactt gcactgggga cagcctattt tgctagtttg ttttgtttcg ttttgttttg
      241 atggagagcg tatgttagtt acgattcaca caaaaaacca acacacagat gtaatgaaaa
      301 taaagatatt ttattgcggc cgctgattta aatgttaatt aactagccat gaccaaaatc
      361 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
      421 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
      481 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
      541 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
      601 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
      661 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
      721 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
      781 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
      841 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
      901 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
      961 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     1021 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttcttaatt
     1081 aacctgcagg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
     1141 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
     1201 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
     1261 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
     1321 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
     1381 tcgctattac catgatgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
     1441 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttgactag
     1501 tcagggcccc aaccccccca agcccccatt tcacaacacg ctggcgctac aggcgcgtga
     1561 cttccccttg ctttggggcg gggggctgag actcctatgt gctccggatt ggtcaggcac
     1621 ggccttcggc cccgcctcct gccaccgcag attggccgct aggcctcccc gagcgccctg
     1681 cctccgaggg ccggcgcacc ataaaagaag ccgccctagc cacgtcccct cgcagttcgg
     1741 cggtcccgcg ggtctgtctc aagcttctgc cttctccctc ctgtgagttt ggtaagtcac
     1801 tgactgtcta tgcctgggaa agggtgggca ggagatgggg cagtgcagga aaagtggcac
     1861 tatgaaccct gcagccctag tttgacaatt aatcattggc atagtatatc tgcatagtat
     1921 aatacaactc actatagcaa ttgtactaac cttcttctct ttcctctcct gacaggagga
     1981 gccatcatgg ccaagttgac cagtgccgtt ccggtgctca ccgcgcgcga cgtcgccgga
     2041 gcggtcgagt tctggaccga ccggctcggg ttctcccggg acttcgtgga ggacgacttc
     2101 gccggtgtgg tccgggacga cgtgaccctg ttcatcagcg cggtccagga ccaggtggtg
     2161 ccggacaaca ccctggcctg ggtgtgggtg cgcggcctgg acgagctgta cgccgagtgg
     2221 tcggaggtcg tgtccacgaa cttccgggac gcctccgggc cggccatgac cgagatcggc
     2281 gagcagccgt gggggcggga gttcgccctg cgcgacccgg ccggcaactg cgtgcacttt
     2341 gtggcagagg agcaggactg aggataagct agaagctcgc tttcttgctg tccaatttct
     2401 attaaaggtt cctttgttcc ctaagtccaa ctactaaact gggggatatt atgaagggcc
     2461 ttgagcatct ggattctgcc taataaaaaa catttatttt cattgcaatg atgtatttaa
     2521 attatttctg aatattttac taaaaaggga atgtgggagg tcagtgcatt taaaacataa
     2581 agaaatgaag agctagttca aaccttggga aaatacacta tatcttaaac tccatgaaag
     2641 aaggtgaggc tgcaaacagc taatgcacat tggcaacagc ccctgatgcc tatgccttat
     2701 tcatccctca gaaaaggatt caagtagagg cttgatttgg aggttaaagt tttgctatgc
     2761 tgtattttag aattaattcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct
     2821 cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt
     2881 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     2941 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     3001 ctggccagct agcactcatt tacccggaga cagggagagg ctcttctgcg tgtagtggtt
     3061 gtgcagagcc tcgtgcatca cggagcatga gaagacgttc ccctgctgcc acctgctctt
     3121 gtccacggtg agcttgctgt agaggaagaa ggagccgtcg gagtccagca cgggaggcgt
     3181 ggtcttgtag ttgttctccg gctgcccatt gctctcccac tccacggcga tgtcgctggg
     3241 atagaagcct ttgaccaggc aggtcaggct gacctggttc ttggtcatct cctcccggga
     3301 tgggggcagg gtgtacacct gtggttctcg gggctgccct ttggctttgg agatggtttt
     3361 ctcgatgggg gctgggaggg ctttgttgga gaccttgcac ttgtactcct tgccattcag
     3421 ccagtcctgg tgcaggacgg tgaggacgct gaccacacgg tacgtgctgt tgtactgctc
     3481 ctcccgcggc tttgtcttgg cattatgcac ctccacgccg tccacgtacc agttgaactt
     3541 gacctcaggg tcttcgtggc tcacgtccac caccacgcat gtgacctcag gggtccggga
     3601 gatcatgagg gtgtccttgg gttttggggg gaagaggaag actgacggtc cccccaggag
     3661 ttcaggtgct gggcacggtg ggcatgtgtg agttttgtca gatcttcact tgtcgtcatc
     3721 gtctttgtag tcagaaccag agtgcccttc tgatcgggga acttcggaat cgggagagag
     3781 ctcagagatt gagtgtgacg ttgctgatcg aggagtggag ggtcccaaaa cggaggccgt
     3841 cgcggaagga gttgctgtgg tgtgggggcc aagcggtgtg ctgctggact gaacagaaga
     3901 cagggctgaa ctattgaagg aagcaaggat ggaggacagt atatccagtt gctctgtaga
     3961 gggaccatgg gacgggcctg agacggagtc ttctctcgct ctcaagagct ggggcagcgg
     4021 acccagggcc tcccttgacg ggtggcgaga aggtacttgg tcgagttgtt ctctactatt
     4081 gctggaccta ctgagtgagt caagcggcct tgaggggaga agcggatcgc gagagagttg
     4141 ccgttgtggt gacaggggaa gcgggggtgg ttgggggtcg agatctcgtg ttcggcgcgg
     4201 aggtggaagt gtggagagcc actctcttgg agcgcccagg tcaagcgcgt ctctactgcc
     4261 aaaccgccct gccatggccc cagggtagtc tctgggtggt tgtcgccgtg gaagcgtact
     4321 gcccctgtct ttaggctcca gtcgcccggg cgccgcgtcc atgcactcct gagagccggg
     4381 cctgctcaaa ggtctcaaga caggcgctgg ggcttcttca agccgttcgc gtgacagttg
     4441 tcttctgagc aggaggtcca actgttctcg gcttgaatag cgtgatgcag gctgggagag
     4501 gcttcccgtg cctcctcccg cataaatccg gccgtaggac gccgcgggta ccgcccggtg
     4561 gccgagagtc gctgcccggc accaagaaag ctccggtgct ccgcgggtct gaggcaccag
     4621 tggatactgc aagcgattct taagaatgcc tttgcgttgt cgttgtgccc taccgagaga
     4681 tgtttcatca ccgctagtaa gagccgggtc aggctgatta ttgttcgcgg cgtctttgct
     4741 cgtatcgagg ccaagccggc gctcacttcc ccatgtttga agtgcacatg gtgctgcctc
     4801 acactctcca agagcaggcc aataagacaa cagatcgttt ccgtctacgt ccttaggatg
     4861 ggtcaagagt ctctctgatt gcgcggcgcg acagagcggt ctctggaacc gacccctggt
     4921 tctaccgttg tcttcacttt ctgaagatgg aatagaaaga gaccgctcct cttcaagtga
     4981 caagtcggaa tcgctatcac tgtcggcacc agcgtcgcgg tggaacatcg caacgtcgag
     5041 gtcagtgggt ccagcatggg cttggagaga atgatcggtg tggtcagctg cgctaccatg
     5101 cctcacgaga acgttatccc taaggtaact gcggccgcgt tgtgagtctt gatcttgggt
     5161 ccggccagac cgtgctgagc ttacacttga aacagttgac gcgccaagag ttatccgtat
     5221 aagcccactt tcttcgaaca aggcggtgtt gttgtatgcc ccaggtccca ggcctggtgc
     5281 cggtctagcc tcttctggtg cagccttgcg tccaaggcaa gccggcatcc atgctgctct
     5341 ggcgtcggcg ttaagaacgc agaacagcag aagcactgcc aatccttgga gtccacacaa
     5401 ccctgcgtgc agatagtgaa aggccagaat tgaatggttg acggccagca aaccaaacaa
     5461 ccaggacgcg cttacaagaa gcagcaaaag aaagcttgat cgcaatgtca gcgcgctcgt
     5521 cttttttgct tccctttggc cagtactaca agaggtgcga gcagccagaa ggaacatagt
     5581 accgttcatt acaatcacca agactaccgg accagcaaaa gaccatatga gcggctcatg
     5641 aacactaatc caacagaagt cagggtttcc gtagccctcc gggtccaacc ccaccgcaag
     5701 tccaaggagg acagcgggca caccccaccc caaggcatga taaaacctca ttgcacccct
     5761 atctacgttc ctaggctcaa cttgcattcg gtacagatga agcccctgca caaacagcca
     5821 agcgaaggtg ctaagaaaga aataatgaag gaggatggca accgccgtgc aaacaagctg
     5881 attgtgcgtt cgatgaatcc caaggagaaa caggagttcg gctaccccaa gagccgcagc
     5941 aacgttggcg tgtatgcccc gaacattgct tttcagactg cgcaaggaaa gcagaatcgc
     6001 cgcggtaagc accagggcgg ctacggatac ggccacaact acatgtgtaa atacggccag
     6061 cagttccagg tccccctcca agcgttccct tgggcttgca tccatcaata caccgaaggt
     6121 gccggtttgg aagtaaaggt tctcgttggt ctgcgcatct ttcagcgctt catccacggt
     6181 ctggcggccg ctcgccgcgt taatcaccgc ggtgcgcacc gcataccaaa acgcgctcat
     6241 ctgcggaatg ttcggcataa tttcgccttt ctgcgcgttt tccatggtcg ccgcaatgcg
     6301 cggatctttc gccagttctt cttcatagct tttcagcgcc accgcgccca gcggtttatc
     6361 tttgttcacc gcttccaggc cttcatcggt cagcagatag ttttccagaa attctttcgc
     6421 cagttctttg ttcgggctcg ccgcgttaat gcccgcgctc agcacgccca caaacggttt
     6481 gctcggctgg cctttaaagg tcggcagcac ggtcacgcca tagttcactt tgctggtatc
     6541 aatgttgctc cacgcccacg ggccgttaat ggtcatcgcg gtttcgcctt tgttaaacgc
     6601 cgcttccgca atgctataat cggtatccgc gttcatatgt ttgtttttaa tcagatccac
     6661 cagaaaggtc aggcccgctt tcgcgcccgc gttatccacg cccacatctt taatatcata
     6721 tttgccgttt tcatatttaa acgcatagcc gccatccgcc gcaatcagcg gccaggtaaa
     6781 atacggttcc tgcaggttaa acatcagcgc gcttttgcct ttcgctttca gttctttatc
     6841 cagcgccgga atttcttccc aggttttcgg cgggttcggc agcagatctt tgttataaat
     6901 caggctcagc gcttccaccg caatcggata cgcaatcagt ttgccgttat agcgcaccgc
     6961 atcccaggta aacggataca gtttatcctg aaacgcttta tccggggtaa tttccgccag
     7021 caggccgctc tgcgcatagc cgccaaagcg atcatgcgcc caaaaaataa tatccgggcc
     7081 atcgccggtc gccgccacct gcggaaattt ttcttccagt ttatccggat gttccacggt
     7141 cactttaatg ccggtatctt tttcaaattt tttgcccact tccgccaggc cgttatagcc
     7201 tttatcgccg ttaatccaaa tcaccagttt gccttcttca attttggatc cagcgtaatc
     7261 tggaacatcg tatgggtacg aattcgtgac aagtgcaaga cttagtgcaa tgcaagacag
     7321 gagttgcatc ctgtacatgg tggccctcct tcgccggtga tctcaggtag gcgccggtca
     7381 cagcttggat ctgtaacggc gcagaacaga aaacgaaaca aagacgtaga gttgagcaag
     7441 cagggtcagg caaagcgtgg agagccggct gagtctaggt aggctccaag ggagcgccgg
     7501 acaaaggccc ggtctcgacc tgagctttaa acttacctag acggcggacg cagttcagga
     7561 ggcaccacag gcgggaggcg gcagaacgcg actcaaccgg cgtggatggc ggcctcaggt
     7621 agggcggcgg gcgcgtgaag gagagatgcg agcccctcga agcttcagct gtgttctggc
     7681 ggcaaacccg ttgcgaaaaa gaacgttcac ggcgactact gcacttatat acggttctcc
     7741 cccaccctcg gg
//
