LOCUS       sequence-442646-        7095 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 7095)
  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 7095)
  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-ADGRA3 CTFdeltaTA-FLAG
FEATURES             Location/Qualifiers
     source          1..7095
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    368..762
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     polyA_signal    866..987
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(903..922)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     957..976
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(1702..1725)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(4588..4614)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     sig_peptide     complement(4615..4674)
                     /codon_start=1
                     /product="human interleukin-2 signal sequence"
                     /label=IL-2 signal sequence
                     /translation="MYRMQLLSCIALSLALVTNS"
     LTR             4724..4992
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     promoter        complement(5005..5216)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     misc_feature    5253..5344
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     complement(5285..5304)
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     polyA_signal    5355..5403
                     /note="synthetic polyadenylation signal"
     rep_origin      5511..6099
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6000..6019
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        6179..6482
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             join(7075..7095,1..354)
                     /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"
ORIGIN
        1 gttccggtgc tcaccgcgcg cgacgtcgcc ggagcggtcg agttctggac cgaccggctc
       61 gggttctccc gggacttcgt ggaggacgac ttcgccggtg tggtccggga cgacgtgacc
      121 ctgttcatca gcgcggtcca ggaccaggtg gtgccggaca acaccctggc ctgggtgtgg
      181 gtgcgcggcc tggacgagct gtacgccgag tggtcggagg tcgtgtccac gaacttccgg
      241 gacgcctccg ggccggccat gaccgagatc ggcgagcagc cgtgggggcg ggagttcgcc
      301 ctgcgcgacc cggccggcaa ctgcgtgcac tttgtggcag aggagcagga ctgaggataa
      361 gctagaagct cgctttcttg ctgtccaatt tctattaaag gttcctttgt tccctaagtc
      421 caactactaa actgggggat attatgaagg gccttgagca tctggattct gcctaataaa
      481 aaacatttat tttcattgca atgatgtatt taaattattt ctgaatattt tactaaaaag
      541 ggaatgtggg aggtcagtgc atttaaaaca taaagaaatg aagagctagt tcaaaccttg
      601 ggaaaataca ctatatctta aactccatga aagaaggtga ggctgcaaac agctaatgca
      661 cattggcaac agcccctgat gcctatgcct tattcatccc tcagaaaagg attcaagtag
      721 aggcttgatt tggaggttaa agttttgcta tgctgtattt tagaattaat tccataccac
      781 atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca
      841 taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata
      901 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
      961 tttgtccaaa ctcatcaatg tatcttatca tgtctggcca gctagcactc atttacccgg
     1021 agacagggag aggctcttct gcgtgtagtg gttgtgcaga gcctcgtgca tcacggagca
     1081 tgagaagacg ttcccctgct gccacctgct cttgtccacg gtgagcttgc tgtagaggaa
     1141 gaaggagccg tcggagtcca gcacgggagg cgtggtcttg tagttgttct ccggctgccc
     1201 attgctctcc cactccacgg cgatgtcgct gggatagaag cctttgacca ggcaggtcag
     1261 gctgacctgg ttcttggtca tctcctcccg ggatgggggc agggtgtaca cctgtggttc
     1321 tcggggctgc cctttggctt tggagatggt tttctcgatg ggggctggga gggctttgtt
     1381 ggagaccttg cacttgtact ccttgccatt cagccagtcc tggtgcagga cggtgaggac
     1441 gctgaccaca cggtacgtgc tgttgtactg ctcctcccgc ggctttgtct tggcattatg
     1501 cacctccacg ccgtccacgt accagttgaa cttgacctca gggtcttcgt ggctcacgtc
     1561 caccaccacg catgtgacct caggggtccg ggagatcatg agggtgtcct tgggttttgg
     1621 ggggaagagg aagactgacg gtccccccag gagttcaggt gctgggcacg gtgggcatgt
     1681 gtgagttttg tcagatcttc acttgtcgtc atcgtctttg tagtcagaac cgacggtggt
     1741 ttcgtgcttc cacaaaccgg tccgaacgtt gcctgtgctg tctgtgccca gcaagggccc
     1801 ctcctgtccg ttggacttga ttggaccgtt ttggatcgcg aggttaagac cgtaactttt
     1861 ctgttggttc tcgagttcaa ccacagcagg cttccgaagg gcgtcctttt tgctggttgt
     1921 gctgactggc ttttcgaagt ttctgctaga ctttggcagg gtactacagg catcagagga
     1981 gtcctgttga gggggattgt actgacgctc acggtatgcg aggtaggccc gacgagacct
     2041 gctgtgcccc tcgttattac cgaggcgact cttgggcagg ccattttgta cggaaccctc
     2101 gacactagtc ggcacatcgt aggcatactc gcggagcaca gtcaggcgag acgctctatg
     2161 gcctttgctc cggttcttgt gatgcctgga agagtgtaca ttagtgcgga attgcacctc
     2221 caatggagcc acgtgcattt taatgtcgtt gtccatggag tgctcagtca gggagttgtc
     2281 cagctgtggg gtgctgttca aaggcagaga gtttgcatgg cactgggcag ccgcggcctg
     2341 aagatttgtc aacttgcacc cctgagatga gtttttgaag ctagaagcag atttgttagt
     2401 gcaactagac tcagcggaag agttcggaca cttgggcgcc tccccgttgg taccgttaga
     2461 gttagggggc tggacattca cctgcacgga atagctagac ctgcccgggc aacaggtcat
     2521 gatccaagcc aaacgaacgt cttcgcggtt cacacagtga tgcacgacga aaaaagcgct
     2581 gaaactcagg gaggtggcac caaacacaaa tgagaacacc aggtccagtg gatagtacag
     2641 ggagacggcc agagcgccga acatccacag cgccacgtac aggagcaggg tcaagctggc
     2701 gcccaacagc tgagagtgga aggtgtgttc gttctcgagg gcagatgtgc tgatcaggga
     2761 gagactcatg ctgtcctggt ggttaatttc tccgttctcg ttggctgcca ggcgctgttg
     2821 ctcttctgtc ggctccttga gctcgtactt acgctcgggg tgtcttttca actgaatgaa
     2881 aatggacaga aagtacatac agttgacgaa tgtgatgaaa gaagctggcc catagaaagc
     2941 gccaagggag ggttcccacg ccatccagca gtatggggcg ttcgggcggg agccgtaatt
     3001 cttgatgtta gcggctgcgg tgatgccgca cacgataata gggataccgc ccccgatcag
     3061 gtaaaaacgc aacattggac ggggaggggg agggggttca tccggatcct ggcagcgctt
     3121 agctttcttt gtgacctgct tgtagatatt gcgggcggtg acccctaccc agagcacagt
     3181 agccagggtg gagtaatgga ggataatgcc aaccgcctgg cagatagaag cgttcctggt
     3241 ctgtgtaatg cccccaacga acacaacgca ggtcagaaag atgtggaaac acagattaac
     3301 cagcatatgc catgacttaa gagagatacg gatgaggctg tgatgataaa tgtagctaac
     3361 gataacagcc agcaagcaca gcaacaggat aatagcagtg gtgtacacaa cggggtgaag
     3421 cagagaagcc gcttgagtgt acagctcgga cccggtcaga tcttggaagt aaaggttctc
     3481 gttggtctgc gcatctttca gcgcttcatc cacggtctgg cggccgctcg ccgcgttaat
     3541 caccgcggtg cgcaccgcat accaaaacgc gctcatctgc ggaatgttcg gcataatttc
     3601 gcctttctgc gcgttttcca tggtcgccgc aatgcgcgga tctttcgcca gttcttcttc
     3661 atagcttttc agcgccaccg cgcccagcgg tttatctttg ttcaccgctt ccaggccttc
     3721 atcggtcagc agatagtttt ccagaaattc tttcgccagt tctttgttcg ggctcgccgc
     3781 gttaatgccc gcgctcagca cgcccacaaa cggtttgctc ggctggcctt taaaggtcgg
     3841 cagcacggtc acgccatagt tcactttgct ggtatcaatg ttgctccacg cccacgggcc
     3901 gttaatggtc atcgcggttt cgcctttgtt aaacgccgct tccgcaatgc tataatcggt
     3961 atccgcgttc atatgtttgt ttttaatcag atccaccaga aaggtcaggc ccgctttcgc
     4021 gcccgcgtta tccacgccca catctttaat atcatatttg ccgttttcat atttaaacgc
     4081 atagccgcca tccgccgcaa tcagcggcca ggtaaaatac ggttcctgca ggttaaacat
     4141 cagcgcgctt ttgcctttcg ctttcagttc tttatccagc gccggaattt cttcccaggt
     4201 tttcggcggg ttcggcagca gatctttgtt ataaatcagg ctcagcgctt ccaccgcaat
     4261 cggatacgca atcagtttgc cgttatagcg caccgcatcc caggtaaacg gatacagttt
     4321 atcctgaaac gctttatccg gggtaatttc cgccagcagg ccgctctgcg catagccgcc
     4381 aaagcgatca tgcgcccaaa aaataatatc cgggccatcg ccggtcgccg ccacctgcgg
     4441 aaatttttct tccagtttat ccggatgttc cacggtcact ttaatgccgg tatctttttc
     4501 aaattttttg cccacttccg ccaggccgtt atagccttta tcgccgttaa tccaaatcac
     4561 cagtttgcct tcttcaattt tggatccagc gtaatctgga acatcgtatg ggtacgaatt
     4621 cgtgacaagt gcaagactta gtgcaatgca agacaggagt tgcatcctgt acatggtggc
     4681 cctccttcgc cggtgatctc aggtaggcgc cggtcacagc ttggatctgt aacggcgcag
     4741 aacagaaaac gaaacaaaga cgtagagttg agcaagcagg gtcaggcaaa gcgtggagag
     4801 ccggctgagt ctaggtaggc tccaagggag cgccggacaa aggcccggtc tcgacctgag
     4861 ctttaaactt acctagacgg cggacgcagt tcaggaggca ccacaggcgg gaggcggcag
     4921 aacgcgactc aaccggcgtg gatggcggcc tcaggtaggg cggcgggcgc gtgaaggaga
     4981 gatgcgagcc cctcgaagct tcagctgtgt tctggcggca aacccgttgc gaaaaagaac
     5041 gttcacggcg actactgcac ttatatacgg ttctccccca ccctcgggaa aaaggcggag
     5101 ccagtacacg acatcacttt cccagtttac cccgcgccac cttctctagg cacccgttca
     5161 attgccgacc cctcccccca acttctcggg gactgtgggc gatgtgcgct ctgcccactg
     5221 acgggcaccg gagcgatcgc agatccttcg atagagaaat gttctggcac ctgcacttgc
     5281 actggggaca gcctattttg ctagtttgtt ttgtttcgtt ttgttttgat ggagagcgta
     5341 tgttagttac gattcacaca aaaaaccaac acacagatgt aatgaaaata aagatatttt
     5401 attgcggccg ctgatttaaa tgttaattaa ctagccatga ccaaaatccc ttaacgtgag
     5461 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     5521 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     5581 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     5641 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     5701 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     5761 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     5821 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     5881 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     5941 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     6001 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     6061 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     6121 ttacggttcc tggccttttg ctggcctttt gctcacatgt tcttaattaa cctgcaggcg
     6181 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
     6241 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
     6301 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
     6361 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
     6421 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
     6481 tgatgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
     6541 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttgactagtc agggccccaa
     6601 cccccccaag cccccatttc acaacacgct ggcgctacag gcgcgtgact tccccttgct
     6661 ttggggcggg gggctgagac tcctatgtgc tccggattgg tcaggcacgg ccttcggccc
     6721 cgcctcctgc caccgcagat tggccgctag gcctccccga gcgccctgcc tccgagggcc
     6781 ggcgcaccat aaaagaagcc gccctagcca cgtcccctcg cagttcggcg gtcccgcggg
     6841 tctgtctcaa gcttctgcct tctccctcct gtgagtttgg taagtcactg actgtctatg
     6901 cctgggaaag ggtgggcagg agatggggca gtgcaggaaa agtggcacta tgaaccctgc
     6961 agccctagtt tgacaattaa tcattggcat agtatatctg catagtataa tacaactcac
     7021 tatagcaatt gtactaacct tcttctcttt cctctcctga caggaggagc catcatggcc
     7081 aagttgacca gtgcc
//
