LOCUS       sequence-430299-        7341 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 7341)
  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 7341)
  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-ADGRB3 CTFdeltaTA-FLAG
FEATURES             Location/Qualifiers
     source          1..7341
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(229..817)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(309..328)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    925..973
                     /note="synthetic polyadenylation signal"
     misc_feature    984..1075
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     1024..1043
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        1112..1323
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             1336..1604
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     sig_peptide     1654..1713
                     /codon_start=1
                     /product="human interleukin-2 signal sequence"
                     /label=IL-2 signal sequence
                     /translation="MYRMQLLSCIALSLALVTNS"
     CDS             1714..1740
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4849..4872
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    5587..5708
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5598..5617)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5652..5671
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     polyA_signal    5812..6206
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     CDS             complement(6220..6594)
                     /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        join(7187..7341,1..149)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gggcagttta ccgtaaatac tccacccatt gacgtcaatg gaaagtccct attggcgtta
       61 ctatgggaac atacgtcatt attgacgtca atgggcgggg gtcgttgggc ggtcagccag
      121 gcgggccatt taccgtaagt tatgtaacgc ctgcaggtta attaagaaca tgtgagcaaa
      181 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
      241 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
      301 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
      361 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
      421 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
      481 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
      541 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
      601 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
      661 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
      721 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
      781 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
      841 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tggctagtta
      901 attaacattt aaatcagcgg ccgcaataaa atatctttat tttcattaca tctgtgtgtt
      961 ggttttttgt gtgaatcgta actaacatac gctctccatc aaaacaaaac gaaacaaaac
     1021 aaactagcaa aataggctgt ccccagtgca agtgcaggtg ccagaacatt tctctatcga
     1081 aggatctgcg atcgctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc
     1141 gagaagttgg ggggaggggt cggcaattga acgggtgcct agagaaggtg gcgcggggta
     1201 aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg
     1261 tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca
     1321 cagctgaagc ttcgaggggc tcgcatctct ccttcacgcg cccgccgccc tacctgaggc
     1381 cgccatccac gccggttgag tcgcgttctg ccgcctcccg cctgtggtgc ctcctgaact
     1441 gcgtccgccg tctaggtaag tttaaagctc aggtcgagac cgggcctttg tccggcgctc
     1501 ccttggagcc tacctagact cagccggctc tccacgcttt gcctgaccct gcttgctcaa
     1561 ctctacgtct ttgtttcgtt ttctgttctg cgccgttaca gatccaagct gtgaccggcg
     1621 cctacctgag atcaccggcg aaggagggcc accatgtaca ggatgcaact cctgtcttgc
     1681 attgcactaa gtcttgcact tgtcacgaat tcgtacccat acgatgttcc agattacgct
     1741 ggatccaaaa ttgaagaagg caaactggtg atttggatta acggcgataa aggctataac
     1801 ggcctggcgg aagtgggcaa aaaatttgaa aaagataccg gcattaaagt gaccgtggaa
     1861 catccggata aactggaaga aaaatttccg caggtggcgg cgaccggcga tggcccggat
     1921 attatttttt gggcgcatga tcgctttggc ggctatgcgc agagcggcct gctggcggaa
     1981 attaccccgg ataaagcgtt tcaggataaa ctgtatccgt ttacctggga tgcggtgcgc
     2041 tataacggca aactgattgc gtatccgatt gcggtggaag cgctgagcct gatttataac
     2101 aaagatctgc tgccgaaccc gccgaaaacc tgggaagaaa ttccggcgct ggataaagaa
     2161 ctgaaagcga aaggcaaaag cgcgctgatg tttaacctgc aggaaccgta ttttacctgg
     2221 ccgctgattg cggcggatgg cggctatgcg tttaaatatg aaaacggcaa atatgatatt
     2281 aaagatgtgg gcgtggataa cgcgggcgcg aaagcgggcc tgacctttct ggtggatctg
     2341 attaaaaaca aacatatgaa cgcggatacc gattatagca ttgcggaagc ggcgtttaac
     2401 aaaggcgaaa ccgcgatgac cattaacggc ccgtgggcgt ggagcaacat tgataccagc
     2461 aaagtgaact atggcgtgac cgtgctgccg acctttaaag gccagccgag caaaccgttt
     2521 gtgggcgtgc tgagcgcggg cattaacgcg gcgagcccga acaaagaact ggcgaaagaa
     2581 tttctggaaa actatctgct gaccgatgaa ggcctggaag cggtgaacaa agataaaccg
     2641 ctgggcgcgg tggcgctgaa aagctatgaa gaagaactgg cgaaagatcc gcgcattgcg
     2701 gcgaccatgg aaaacgcgca gaaaggcgaa attatgccga acattccgca gatgagcgcg
     2761 ttttggtatg cggtgcgcac cgcggtgatt aacgcggcga gcggccgcca gaccgtggat
     2821 gaagcgctga aagatgcgca gaccaacgag aacctttact tccaacagca acctagagaa
     2881 ataatcatgg aatcctctgg cacaccttca gttaccctaa tagtaggcag tggtctttct
     2941 tgcttggcct tgattaccct agcagttgtc tatgcagcat tatggaggta catacgctct
     3001 gagagatcca taatactaat taacttctgc ctgtctatca tctcatccaa tatcctcata
     3061 ctggttggac agactcagac acataataag agtatctgca caaccaccac tgcatttttg
     3121 cactttttct tcctggcttc attctgttgg gttttgactg aggcgtggca atcatatatg
     3181 gctgtaactg gaaaaattag gacacggctt ataagaaaac gctttttgtg ccttggatgg
     3241 ggtttaccag cattagtagt ggccacatca gtaggcttca ccagaacaaa aggatatggc
     3301 actgatcact actgctggct ctctcttgaa ggaggactac tctatgcttt tgtgggacct
     3361 gcagccgctg ttgtcctggt caacatggtg attggcattt tggtatttaa taaacttgtt
     3421 tccagagatg gaatcctaga taaaaagctc aaacacagag ccggtcagat gagtgagcct
     3481 catagcggtt tgacgctcaa atgtgccaag tgtggagtag tttcaacaac agctttgtca
     3541 gccaccaccg ccagtaacgc catggcgtct ctttggagct cctgtgtggt gttgcccctt
     3601 ctggctttga cgtggatgtc tgcggttctg gccatgacag ataaacgctc catattgttt
     3661 caaatacttt ttgctgtgtt tgattcattg caaggctttg ttatagtcat ggtccactgc
     3721 attcttcgga gagaggttca ggatgcattt agatgccgat tgagaaactg tcaggatccc
     3781 atcaatgcag attcttcgag ttcgtttcct aatgggcatg ctcaaatcat gacagacttt
     3841 gaaaaggatg tagacattgc ctgtcgatca gttcttcata aggatattgg tccttgccga
     3901 gcagccacaa taacaggaac actttctagg atttctctaa atgatgatga agaagaaaag
     3961 ggaacaaacc ctgaagggct aagctattca acattgcctg gaaatgtcat ttccaaagtc
     4021 atcatccagc aacccacagg tttgcacatg cccatgagta tgaatgagct tagcaatcca
     4081 tgtttgaaaa aagaaaatag tgaattgcgg agaactgtgt acttatgtac ggatgataat
     4141 ttgagagggg ctgacatgga catagtccat cctcaagaaa gaatgatgga aagtgactat
     4201 attgtgatgc ccagaagttc tgtaaataac cagccttcaa tgaaagaaga aagcaaaatg
     4261 aatattggca tggaaacctt gccgcatgaa aggctattgc actacaaagt aaaccctgaa
     4321 ttcaatatga atccccctgt aatggaccag ttcaatatga acttagagca acatctcgca
     4381 ccccaggaac atatgcagaa tttgcccttt gaacctcgca cagctgtgaa gaatttcatg
     4441 gcctctgagt tggatgataa tgcaggacta tcaagaagtg aaactggatc aacgatatca
     4501 atgagttctt tagagagaag aaaatcacga tattcagacc ttgactttga gaaggtcatg
     4561 catacaagga agaggcatat ggaactattt caagaactaa atcagaaatt tcaaactttg
     4621 gacagatttc gggatatacc aaatacaagc agtatggaaa accccgcacc aaacaagaat
     4681 ccatgggaca ctttcaaaaa ccccagtgaa tacccgcatt acaccacaat caatgtctta
     4741 gacacagagg caaaggatgc tttggaactg aggccagcag agtgggagaa gtgtctgaat
     4801 ttgcctctgg atgtgcaaga gggtgacttt caaacagaag ttggttctga ctacaaagac
     4861 gatgacgaca agtgaagatc tgacaaaact cacacatgcc caccgtgccc agcacctgaa
     4921 ctcctggggg gaccgtcagt cttcctcttc cccccaaaac ccaaggacac cctcatgatc
     4981 tcccggaccc ctgaggtcac atgcgtggtg gtggacgtga gccacgaaga ccctgaggtc
     5041 aagttcaact ggtacgtgga cggcgtggag gtgcataatg ccaagacaaa gccgcgggag
     5101 gagcagtaca acagcacgta ccgtgtggtc agcgtcctca ccgtcctgca ccaggactgg
     5161 ctgaatggca aggagtacaa gtgcaaggtc tccaacaaag ccctcccagc ccccatcgag
     5221 aaaaccatct ccaaagccaa agggcagccc cgagaaccac aggtgtacac cctgccccca
     5281 tcccgggagg agatgaccaa gaaccaggtc agcctgacct gcctggtcaa aggcttctat
     5341 cccagcgaca tcgccgtgga gtgggagagc aatgggcagc cggagaacaa ctacaagacc
     5401 acgcctcccg tgctggactc cgacggctcc ttcttcctct acagcaagct caccgtggac
     5461 aagagcaggt ggcagcaggg gaacgtcttc tcatgctccg tgatgcacga ggctctgcac
     5521 aaccactaca cgcagaagag cctctccctg tctccgggta aatgagtgct agctggccag
     5581 acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat
     5641 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
     5701 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg
     5761 aggtttttta aagcaagtaa aacctctaca aatgtggtat ggaattaatt ctaaaataca
     5821 gcatagcaaa actttaacct ccaaatcaag cctctacttg aatccttttc tgagggatga
     5881 ataaggcata ggcatcaggg gctgttgcca atgtgcatta gctgtttgca gcctcacctt
     5941 ctttcatgga gtttaagata tagtgtattt tcccaaggtt tgaactagct cttcatttct
     6001 ttatgtttta aatgcactga cctcccacat tcccttttta gtaaaatatt cagaaataat
     6061 ttaaatacat cattgcaatg aaaataaatg ttttttatta ggcagaatcc agatgctcaa
     6121 ggcccttcat aatatccccc agtttagtag ttggacttag ggaacaaagg aacctttaat
     6181 agaaattgga cagcaagaaa gcgagcttct agcttatcct cagtcctgct cctctgccac
     6241 aaagtgcacg cagttgccgg ccgggtcgcg cagggcgaac tcccgccccc acggctgctc
     6301 gccgatctcg gtcatggccg gcccggaggc gtcccggaag ttcgtggaca cgacctccga
     6361 ccactcggcg tacagctcgt ccaggccgcg cacccacacc caggccaggg tgttgtccgg
     6421 caccacctgg tcctggaccg cgctgatgaa cagggtcacg tcgtcccgga ccacaccggc
     6481 gaagtcgtcc tccacgaagt cccgggagaa cccgagccgg tcggtccaga actcgaccgc
     6541 tccggcgacg tcgcgcgcgg tgagcaccgg aacggcactg gtcaacttgg ccatgatggc
     6601 tcctcctgtc aggagaggaa agagaagaag gttagtacaa ttgctatagt gagttgtatt
     6661 atactatgca gatatactat gccaatgatt aattgtcaaa ctagggctgc agggttcata
     6721 gtgccacttt tcctgcactg ccccatctcc tgcccaccct ttcccaggca tagacagtca
     6781 gtgacttacc aaactcacag gagggagaag gcagaagctt gagacagacc cgcgggaccg
     6841 ccgaactgcg aggggacgtg gctagggcgg cttcttttat ggtgcgccgg ccctcggagg
     6901 cagggcgctc ggggaggcct agcggccaat ctgcggtggc aggaggcggg gccgaaggcc
     6961 gtgcctgacc aatccggagc acataggagt ctcagccccc cgccccaaag caaggggaag
     7021 tcacgcgcct gtagcgccag cgtgttgtga aatgggggct tgggggggtt ggggccctga
     7081 ctagtcaaaa caaactccca ttgacgtcaa tggggtggag acttggaaat ccccgtgagt
     7141 caaaccgcta tccacgccca ttgatgtact gccaaaaccg catcatcatg gtaatagcga
     7201 tgactaatac gtagatgtac tgccaagtag gaaagtccca taaggtcatg tactgggcat
     7261 aatgccaggc gggccattta ccgtcattga cgtcaatagg gggcgtactt ggcatatgat
     7321 acacttgatg tactgccaag t
//
