LOCUS       sequence-430287-        6297 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 6297)
  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 6297)
  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-ADGRF4 CTF-FLAG
FEATURES             Location/Qualifiers
     source          1..6297
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..48)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     83..102
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(828..851)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(1788..1808)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQM"
     CDS             complement(2916..2942)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     sig_peptide     complement(2943..3002)
                     /codon_start=1
                     /product="human interleukin-2 signal sequence"
                     /label=IL-2 signal sequence
                     /translation="MYRMQLLSCIALSLALVTNS"
     LTR             3052..3320
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     promoter        complement(3333..3544)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     misc_feature    3581..3672
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     complement(3613..3632)
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     polyA_signal    3683..3731
                     /note="synthetic polyadenylation signal"
     rep_origin      3839..4427
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4328..4347
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        4507..4810
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             5403..5777
                     /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    5791..6185
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     polyA_signal    join(6289..6297,1..113)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
       61 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
      121 tggccagcta gcactcattt acccggagac agggagaggc tcttctgcgt gtagtggttg
      181 tgcagagcct cgtgcatcac ggagcatgag aagacgttcc cctgctgcca cctgctcttg
      241 tccacggtga gcttgctgta gaggaagaag gagccgtcgg agtccagcac gggaggcgtg
      301 gtcttgtagt tgttctccgg ctgcccattg ctctcccact ccacggcgat gtcgctggga
      361 tagaagcctt tgaccaggca ggtcaggctg acctggttct tggtcatctc ctcccgggat
      421 gggggcaggg tgtacacctg tggttctcgg ggctgccctt tggctttgga gatggttttc
      481 tcgatggggg ctgggagggc tttgttggag accttgcact tgtactcctt gccattcagc
      541 cagtcctggt gcaggacggt gaggacgctg accacacggt acgtgctgtt gtactgctcc
      601 tcccgcggct ttgtcttggc attatgcacc tccacgccgt ccacgtacca gttgaacttg
      661 acctcagggt cttcgtggct cacgtccacc accacgcatg tgacctcagg ggtccgggag
      721 atcatgaggg tgtccttggg ttttgggggg aagaggaaga ctgacggtcc ccccaggagt
      781 tcaggtgctg ggcacggtgg gcatgtgtga gttttgtcag atcttcactt gtcgtcatcg
      841 tctttgtagt cagaacctcc ctggcgattc atcagctttg agccgttagt ggggcccaga
      901 gatgcgtttt cagctgctct agatttcccc ttcaggctag acattctcat gcgcagagcg
      961 tcgcggatct tgtggtccat gatcgttccg aaaagcagaa tgaaaaatcc ttggaaagca
     1021 ttcaggaggg caaagataat gtggaaggtc aagctggtgc cctcaatcag ggtagcgatg
     1081 ccgaaacccc aggtaagccc aagcagcggg gtcaggatcg cgacattctt gctgatcctc
     1141 ataatgatca caacgtcctg gctcttggag cttccaatgg aggggcgctg agtgttcacg
     1201 gccactacca ggaccacaat caagttcacc gcaacgataa caaaggcagg gatggcgaaa
     1261 gccagcaagg ctttggtgtt gtcccagttc agccagcagg cctcaggacg catgtatccc
     1321 ttttcgggtt cggtgatggc aacggttgtc acggcgataa tcagtggaca gccgtatccg
     1381 atagcaaagc cgatgaccat catgcgactc ttcatcatgc gcctgaaaat gaccaggatt
     1441 ccgtagataa tcagaagcgc cttgaacagc atccagaaaa acagagacag atagaaaaaa
     1501 tgggagaaaa aggtcactgc cacgcacata ttgtaatcct gggccttgat gttaaaatga
     1561 gaaccgataa tgaaccacac atttgcggtc agcaaagaca cggcaatgtt cacgatgcac
     1621 acatgccgca tgtaggaaat ctcagtcact acgacccggc tccacacggt ggcttcgata
     1681 atcaaacaga gcaccaggct caggatggac acggacagac caatgcaggt gatgtaatcc
     1741 agcaccttat cggtcatgga cttggaagac atcagaatag aaaaggacat ttggaagtaa
     1801 aggttctcgt tggtctgcgc atctttcagc gcttcatcca cggtctggcg gccgctcgcc
     1861 gcgttaatca ccgcggtgcg caccgcatac caaaacgcgc tcatctgcgg aatgttcggc
     1921 ataatttcgc ctttctgcgc gttttccatg gtcgccgcaa tgcgcggatc tttcgccagt
     1981 tcttcttcat agcttttcag cgccaccgcg cccagcggtt tatctttgtt caccgcttcc
     2041 aggccttcat cggtcagcag atagttttcc agaaattctt tcgccagttc tttgttcggg
     2101 ctcgccgcgt taatgcccgc gctcagcacg cccacaaacg gtttgctcgg ctggccttta
     2161 aaggtcggca gcacggtcac gccatagttc actttgctgg tatcaatgtt gctccacgcc
     2221 cacgggccgt taatggtcat cgcggtttcg cctttgttaa acgccgcttc cgcaatgcta
     2281 taatcggtat ccgcgttcat atgtttgttt ttaatcagat ccaccagaaa ggtcaggccc
     2341 gctttcgcgc ccgcgttatc cacgcccaca tctttaatat catatttgcc gttttcatat
     2401 ttaaacgcat agccgccatc cgccgcaatc agcggccagg taaaatacgg ttcctgcagg
     2461 ttaaacatca gcgcgctttt gcctttcgct ttcagttctt tatccagcgc cggaatttct
     2521 tcccaggttt tcggcgggtt cggcagcaga tctttgttat aaatcaggct cagcgcttcc
     2581 accgcaatcg gatacgcaat cagtttgccg ttatagcgca ccgcatccca ggtaaacgga
     2641 tacagtttat cctgaaacgc tttatccggg gtaatttccg ccagcaggcc gctctgcgca
     2701 tagccgccaa agcgatcatg cgcccaaaaa ataatatccg ggccatcgcc ggtcgccgcc
     2761 acctgcggaa atttttcttc cagtttatcc ggatgttcca cggtcacttt aatgccggta
     2821 tctttttcaa attttttgcc cacttccgcc aggccgttat agcctttatc gccgttaatc
     2881 caaatcacca gtttgccttc ttcaattttg gatccagcgt aatctggaac atcgtatggg
     2941 tacgaattcg tgacaagtgc aagacttagt gcaatgcaag acaggagttg catcctgtac
     3001 atggtggccc tccttcgccg gtgatctcag gtaggcgccg gtcacagctt ggatctgtaa
     3061 cggcgcagaa cagaaaacga aacaaagacg tagagttgag caagcagggt caggcaaagc
     3121 gtggagagcc ggctgagtct aggtaggctc caagggagcg ccggacaaag gcccggtctc
     3181 gacctgagct ttaaacttac ctagacggcg gacgcagttc aggaggcacc acaggcggga
     3241 ggcggcagaa cgcgactcaa ccggcgtgga tggcggcctc aggtagggcg gcgggcgcgt
     3301 gaaggagaga tgcgagcccc tcgaagcttc agctgtgttc tggcggcaaa cccgttgcga
     3361 aaaagaacgt tcacggcgac tactgcactt atatacggtt ctcccccacc ctcgggaaaa
     3421 aggcggagcc agtacacgac atcactttcc cagtttaccc cgcgccacct tctctaggca
     3481 cccgttcaat tgccgacccc tccccccaac ttctcgggga ctgtgggcga tgtgcgctct
     3541 gcccactgac gggcaccgga gcgatcgcag atccttcgat agagaaatgt tctggcacct
     3601 gcacttgcac tggggacagc ctattttgct agtttgtttt gtttcgtttt gttttgatgg
     3661 agagcgtatg ttagttacga ttcacacaaa aaaccaacac acagatgtaa tgaaaataaa
     3721 gatattttat tgcggccgct gatttaaatg ttaattaact agccatgacc aaaatccctt
     3781 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     3841 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     3901 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     3961 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     4021 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     4081 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     4141 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     4201 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     4261 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     4321 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     4381 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     4441 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc ttaattaacc
     4501 tgcaggcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
     4561 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
     4621 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
     4681 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
     4741 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
     4801 tattaccatg atgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
     4861 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt gactagtcag
     4921 ggccccaacc cccccaagcc cccatttcac aacacgctgg cgctacaggc gcgtgacttc
     4981 cccttgcttt ggggcggggg gctgagactc ctatgtgctc cggattggtc aggcacggcc
     5041 ttcggccccg cctcctgcca ccgcagattg gccgctaggc ctccccgagc gccctgcctc
     5101 cgagggccgg cgcaccataa aagaagccgc cctagccacg tcccctcgca gttcggcggt
     5161 cccgcgggtc tgtctcaagc ttctgccttc tccctcctgt gagtttggta agtcactgac
     5221 tgtctatgcc tgggaaaggg tgggcaggag atggggcagt gcaggaaaag tggcactatg
     5281 aaccctgcag ccctagtttg acaattaatc attggcatag tatatctgca tagtataata
     5341 caactcacta tagcaattgt actaaccttc ttctctttcc tctcctgaca ggaggagcca
     5401 tcatggccaa gttgaccagt gccgttccgg tgctcaccgc gcgcgacgtc gccggagcgg
     5461 tcgagttctg gaccgaccgg ctcgggttct cccgggactt cgtggaggac gacttcgccg
     5521 gtgtggtccg ggacgacgtg accctgttca tcagcgcggt ccaggaccag gtggtgccgg
     5581 acaacaccct ggcctgggtg tgggtgcgcg gcctggacga gctgtacgcc gagtggtcgg
     5641 aggtcgtgtc cacgaacttc cgggacgcct ccgggccggc catgaccgag atcggcgagc
     5701 agccgtgggg gcgggagttc gccctgcgcg acccggccgg caactgcgtg cactttgtgg
     5761 cagaggagca ggactgagga taagctagaa gctcgctttc ttgctgtcca atttctatta
     5821 aaggttcctt tgttccctaa gtccaactac taaactgggg gatattatga agggccttga
     5881 gcatctggat tctgcctaat aaaaaacatt tattttcatt gcaatgatgt atttaaatta
     5941 tttctgaata ttttactaaa aagggaatgt gggaggtcag tgcatttaaa acataaagaa
     6001 atgaagagct agttcaaacc ttgggaaaat acactatatc ttaaactcca tgaaagaagg
     6061 tgaggctgca aacagctaat gcacattggc aacagcccct gatgcctatg ccttattcat
     6121 ccctcagaaa aggattcaag tagaggcttg atttggaggt taaagttttg ctatgctgta
     6181 ttttagaatt aattccatac cacatttgta gaggttttac ttgctttaaa aaacctccca
     6241 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgttt
//
