LOCUS       sequence-429976-        6597 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 6597)
  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 6597)
  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-ADGRG2 CTF-FLAG
FEATURES             Location/Qualifiers
     source          1..6597
                     /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             3024..3047
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    3762..3883
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3773..3792)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3827..3846
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     polyA_signal    3987..4381
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     CDS             complement(4395..4769)
                     /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        5362..5665
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(5745..6333)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5825..5844)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    6441..6489
                     /note="synthetic polyadenylation signal"
     misc_feature    6500..6591
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     6540..6559
                     /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 ccaaacttct ttcggggtcc
     1801 tgctcgatct gtctaggaca tctgtgctgc cagcccagat gatggccctg acatttatca
     1861 cttacattgg ctgcggtctg tctagcattt tcctgtccgt tactctggtg acctacattg
     1921 ccttcgaaaa gatcagaagg gactatccct ctaagatcct catccagctg tgcgctgccc
     1981 tgcttctcct gaacctggtt ttccttctgg actcctggat cgccctctac aagatgcagg
     2041 gcctgtgtat ctctgtggct gttttcctgc actactttct gctcgtgtcc ttcacttgga
     2101 tgggactgga ggctttccac atgtacctgg ccctggtgaa ggtgttcaat acatacatcc
     2161 gcaaatatat cctgaagttc tgcattgtgg gctggggcgt gcctgctgtg gtcgtgacca
     2221 ttatcttgac aattagtccg gataattacg gtctgggctc ctatggtaag tttccaaacg
     2281 ggagccccga tgacttctgc tggatcaaca ataacgctgt attttatatc accgttgtgg
     2341 ggtatttttg cgtgatcttt ctcctgaacg tttccatgtt catcgtcgtg ctggtccaac
     2401 tctgtcgtat caaaaagaaa aagcagctgg gagcacagcg taaaacttca atccaggacc
     2461 tgagatctat cgccggcctg accttcctcc tggggatcac ctggggcttt gcctttttcg
     2521 catggggccc cgtcaacgta acattcatgt acctgtttgc gatctttaac acactgcaag
     2581 gttttttcat cttcatcttc tactgcgtcg ccaaagagaa cgtccgtaag cagtggcgca
     2641 ggtacctgtg ctgtggcaag ctgcgcctcg cagagaactc cgactggtcc aagacagcaa
     2701 ctaacggtct gaagaaacag actgtaaatc agggggtcag ttccagttcc aactccctcc
     2761 aaagcagtag caacagtacc aacagcacca cacttctcgt gaataacgac tgcagtgttc
     2821 acgccagcgg caatggaaac gctagcactg agagaaacgg cgtttctttc tccgtgcaga
     2881 atggcgatgt ttgcctgcac gactttacag ggaagcagca catgttcaat gaaaaggaag
     2941 attcctgtaa cgggaaaggt aggatggctc tgagacgtac ctccaagagg ggtagcctgc
     3001 acttcatcga gcagatgggt tctgactaca aagacgatga cgacaagtga agatctgaca
     3061 aaactcacac atgcccaccg tgcccagcac ctgaactcct ggggggaccg tcagtcttcc
     3121 tcttcccccc aaaacccaag gacaccctca tgatctcccg gacccctgag gtcacatgcg
     3181 tggtggtgga cgtgagccac gaagaccctg aggtcaagtt caactggtac gtggacggcg
     3241 tggaggtgca taatgccaag acaaagccgc gggaggagca gtacaacagc acgtaccgtg
     3301 tggtcagcgt cctcaccgtc ctgcaccagg actggctgaa tggcaaggag tacaagtgca
     3361 aggtctccaa caaagccctc ccagccccca tcgagaaaac catctccaaa gccaaagggc
     3421 agccccgaga accacaggtg tacaccctgc ccccatcccg ggaggagatg accaagaacc
     3481 aggtcagcct gacctgcctg gtcaaaggct tctatcccag cgacatcgcc gtggagtggg
     3541 agagcaatgg gcagccggag aacaactaca agaccacgcc tcccgtgctg gactccgacg
     3601 gctccttctt cctctacagc aagctcaccg tggacaagag caggtggcag caggggaacg
     3661 tcttctcatg ctccgtgatg cacgaggctc tgcacaacca ctacacgcag aagagcctct
     3721 ccctgtctcc gggtaaatga gtgctagctg gccagacatg ataagataca ttgatgagtt
     3781 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     3841 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     3901 tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct
     3961 ctacaaatgt ggtatggaat taattctaaa atacagcata gcaaaacttt aacctccaaa
     4021 tcaagcctct acttgaatcc ttttctgagg gatgaataag gcataggcat caggggctgt
     4081 tgccaatgtg cattagctgt ttgcagcctc accttctttc atggagttta agatatagtg
     4141 tattttccca aggtttgaac tagctcttca tttctttatg ttttaaatgc actgacctcc
     4201 cacattccct ttttagtaaa atattcagaa ataatttaaa tacatcattg caatgaaaat
     4261 aaatgttttt tattaggcag aatccagatg ctcaaggccc ttcataatat cccccagttt
     4321 agtagttgga cttagggaac aaaggaacct ttaatagaaa ttggacagca agaaagcgag
     4381 cttctagctt atcctcagtc ctgctcctct gccacaaagt gcacgcagtt gccggccggg
     4441 tcgcgcaggg cgaactcccg cccccacggc tgctcgccga tctcggtcat ggccggcccg
     4501 gaggcgtccc ggaagttcgt ggacacgacc tccgaccact cggcgtacag ctcgtccagg
     4561 ccgcgcaccc acacccaggc cagggtgttg tccggcacca cctggtcctg gaccgcgctg
     4621 atgaacaggg tcacgtcgtc ccggaccaca ccggcgaagt cgtcctccac gaagtcccgg
     4681 gagaacccga gccggtcggt ccagaactcg accgctccgg cgacgtcgcg cgcggtgagc
     4741 accggaacgg cactggtcaa cttggccatg atggctcctc ctgtcaggag aggaaagaga
     4801 agaaggttag tacaattgct atagtgagtt gtattatact atgcagatat actatgccaa
     4861 tgattaattg tcaaactagg gctgcagggt tcatagtgcc acttttcctg cactgcccca
     4921 tctcctgccc accctttccc aggcatagac agtcagtgac ttaccaaact cacaggaggg
     4981 agaaggcaga agcttgagac agacccgcgg gaccgccgaa ctgcgagggg acgtggctag
     5041 ggcggcttct tttatggtgc gccggccctc ggaggcaggg cgctcgggga ggcctagcgg
     5101 ccaatctgcg gtggcaggag gcggggccga aggccgtgcc tgaccaatcc ggagcacata
     5161 ggagtctcag ccccccgccc caaagcaagg ggaagtcacg cgcctgtagc gccagcgtgt
     5221 tgtgaaatgg gggcttgggg gggttggggc cctgactagt caaaacaaac tcccattgac
     5281 gtcaatgggg tggagacttg gaaatccccg tgagtcaaac cgctatccac gcccattgat
     5341 gtactgccaa aaccgcatca tcatggtaat agcgatgact aatacgtaga tgtactgcca
     5401 agtaggaaag tcccataagg tcatgtactg ggcataatgc caggcgggcc atttaccgtc
     5461 attgacgtca atagggggcg tacttggcat atgatacact tgatgtactg ccaagtgggc
     5521 agtttaccgt aaatactcca cccattgacg tcaatggaaa gtccctattg gcgttactat
     5581 gggaacatac gtcattattg acgtcaatgg gcgggggtcg ttgggcggtc agccaggcgg
     5641 gccatttacc gtaagttatg taacgcctgc aggttaatta agaacatgtg agcaaaaggc
     5701 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     5761 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     5821 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     5881 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     5941 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     6001 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     6061 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     6121 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     6181 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     6241 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     6301 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     6361 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatggc tagttaatta
     6421 acatttaaat cagcggccgc aataaaatat ctttattttc attacatctg tgtgttggtt
     6481 ttttgtgtga atcgtaacta acatacgctc tccatcaaaa caaaacgaaa caaaacaaac
     6541 tagcaaaata ggctgtcccc agtgcaagtg caggtgccag aacatttctc tatcgaa
//
