LOCUS       sequence-429979-        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-ADGRG1 CTF-FLAG
FEATURES             Location/Qualifiers
     source          1..6297
                     /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"
     regulatory      2406..2415
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2724..2747
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    3462..3583
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3473..3492)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3527..3546
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     polyA_signal    3687..4081
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     CDS             complement(4095..4469)
                     /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        5062..5365
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(5445..6033)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5525..5544)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    6141..6189
                     /note="synthetic polyadenylation signal"
     misc_feature    6200..6291
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     6240..6259
                     /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 ccaaacctac tttgcagtgc
     1801 tgatggtctc ctcggtggag gtggacgccg tgcacaagca ctacctgagc ctcctctcct
     1861 acgtgggctg tgtcgtctct gccctggcct gccttgtcac cattgccgcc tacctctgct
     1921 ccagggtgcc cctgccgtgc aggaggaaac ctcgggacta caccatcaag gtgcacatga
     1981 acctgctgct ggccgtcttc ctgctggaca cgagcttcct gctcagcgag ccggtggccc
     2041 tgacaggctc tgaggctggc tgccgagcca gtgccatctt cctgcacttc tccctgctca
     2101 cctgcctttc ctggatgggc ctcgaggggt acaacctcta ccgactcgtg gtggaggtct
     2161 ttggcaccta tgtccctggc tacctactca agctgagcgc catgggctgg ggcttcccca
     2221 tctttctggt gacgctggtg gccctggtgg atgtggacaa ctatggcccc atcatcttgg
     2281 ctgtgcatag gactccagag ggcgtcatct acccttccat gtgctggatc cgggactccc
     2341 tggtcagcta catcaccaac ctgggcctct tcagcctggt gtttctgttc aacatggcca
     2401 tgctagccac catggtggtg cagatcctgc ggctgcgccc ccacacccaa aagtggtcac
     2461 atgtgctgac actgctgggc ctcagcctgg tccttggcct gccctgggcc ttgatcttct
     2521 tctcctttgc ttctggcacc ttccagcttg tcgtcctcta ccttttcagc atcatcacct
     2581 ccttccaagg cttcctcatc ttcatctggt actggtccat gcggctgcag gcccggggtg
     2641 gcccctcccc tctgaagagc aactcagaca gcgccaggct ccccatcagc tcgggcagca
     2701 cctcgtccag ccgcatcggt tctgactaca aagacgatga cgacaagtga agatctgaca
     2761 aaactcacac atgcccaccg tgcccagcac ctgaactcct ggggggaccg tcagtcttcc
     2821 tcttcccccc aaaacccaag gacaccctca tgatctcccg gacccctgag gtcacatgcg
     2881 tggtggtgga cgtgagccac gaagaccctg aggtcaagtt caactggtac gtggacggcg
     2941 tggaggtgca taatgccaag acaaagccgc gggaggagca gtacaacagc acgtaccgtg
     3001 tggtcagcgt cctcaccgtc ctgcaccagg actggctgaa tggcaaggag tacaagtgca
     3061 aggtctccaa caaagccctc ccagccccca tcgagaaaac catctccaaa gccaaagggc
     3121 agccccgaga accacaggtg tacaccctgc ccccatcccg ggaggagatg accaagaacc
     3181 aggtcagcct gacctgcctg gtcaaaggct tctatcccag cgacatcgcc gtggagtggg
     3241 agagcaatgg gcagccggag aacaactaca agaccacgcc tcccgtgctg gactccgacg
     3301 gctccttctt cctctacagc aagctcaccg tggacaagag caggtggcag caggggaacg
     3361 tcttctcatg ctccgtgatg cacgaggctc tgcacaacca ctacacgcag aagagcctct
     3421 ccctgtctcc gggtaaatga gtgctagctg gccagacatg ataagataca ttgatgagtt
     3481 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     3541 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     3601 tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct
     3661 ctacaaatgt ggtatggaat taattctaaa atacagcata gcaaaacttt aacctccaaa
     3721 tcaagcctct acttgaatcc ttttctgagg gatgaataag gcataggcat caggggctgt
     3781 tgccaatgtg cattagctgt ttgcagcctc accttctttc atggagttta agatatagtg
     3841 tattttccca aggtttgaac tagctcttca tttctttatg ttttaaatgc actgacctcc
     3901 cacattccct ttttagtaaa atattcagaa ataatttaaa tacatcattg caatgaaaat
     3961 aaatgttttt tattaggcag aatccagatg ctcaaggccc ttcataatat cccccagttt
     4021 agtagttgga cttagggaac aaaggaacct ttaatagaaa ttggacagca agaaagcgag
     4081 cttctagctt atcctcagtc ctgctcctct gccacaaagt gcacgcagtt gccggccggg
     4141 tcgcgcaggg cgaactcccg cccccacggc tgctcgccga tctcggtcat ggccggcccg
     4201 gaggcgtccc ggaagttcgt ggacacgacc tccgaccact cggcgtacag ctcgtccagg
     4261 ccgcgcaccc acacccaggc cagggtgttg tccggcacca cctggtcctg gaccgcgctg
     4321 atgaacaggg tcacgtcgtc ccggaccaca ccggcgaagt cgtcctccac gaagtcccgg
     4381 gagaacccga gccggtcggt ccagaactcg accgctccgg cgacgtcgcg cgcggtgagc
     4441 accggaacgg cactggtcaa cttggccatg atggctcctc ctgtcaggag aggaaagaga
     4501 agaaggttag tacaattgct atagtgagtt gtattatact atgcagatat actatgccaa
     4561 tgattaattg tcaaactagg gctgcagggt tcatagtgcc acttttcctg cactgcccca
     4621 tctcctgccc accctttccc aggcatagac agtcagtgac ttaccaaact cacaggaggg
     4681 agaaggcaga agcttgagac agacccgcgg gaccgccgaa ctgcgagggg acgtggctag
     4741 ggcggcttct tttatggtgc gccggccctc ggaggcaggg cgctcgggga ggcctagcgg
     4801 ccaatctgcg gtggcaggag gcggggccga aggccgtgcc tgaccaatcc ggagcacata
     4861 ggagtctcag ccccccgccc caaagcaagg ggaagtcacg cgcctgtagc gccagcgtgt
     4921 tgtgaaatgg gggcttgggg gggttggggc cctgactagt caaaacaaac tcccattgac
     4981 gtcaatgggg tggagacttg gaaatccccg tgagtcaaac cgctatccac gcccattgat
     5041 gtactgccaa aaccgcatca tcatggtaat agcgatgact aatacgtaga tgtactgcca
     5101 agtaggaaag tcccataagg tcatgtactg ggcataatgc caggcgggcc atttaccgtc
     5161 attgacgtca atagggggcg tacttggcat atgatacact tgatgtactg ccaagtgggc
     5221 agtttaccgt aaatactcca cccattgacg tcaatggaaa gtccctattg gcgttactat
     5281 gggaacatac gtcattattg acgtcaatgg gcgggggtcg ttgggcggtc agccaggcgg
     5341 gccatttacc gtaagttatg taacgcctgc aggttaatta agaacatgtg agcaaaaggc
     5401 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     5461 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     5521 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     5581 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     5641 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     5701 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     5761 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     5821 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     5881 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     5941 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     6001 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     6061 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatggc tagttaatta
     6121 acatttaaat cagcggccgc aataaaatat ctttattttc attacatctg tgtgttggtt
     6181 ttttgtgtga atcgtaacta acatacgctc tccatcaaaa caaaacgaaa caaaacaaac
     6241 tagcaaaata ggctgtcccc agtgcaagtg caggtgccag aacatttctc tatcgaa
//
