LOCUS       sequence-429985-        6432 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 6432)
  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 6432)
  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-ADGRF5 CTF-FLAG
FEATURES             Location/Qualifiers
     source          1..6432
                     /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             2859..2882
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    3597..3718
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3608..3627)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3662..3681
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     polyA_signal    3822..4216
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     CDS             complement(4230..4604)
                     /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        5197..5500
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(5580..6168)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5660..5679)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    6276..6324
                     /note="synthetic polyadenylation signal"
     misc_feature    6335..6426
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     6375..6394
                     /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 ccaaacatca ttctccatcc
     1801 tcatgtcccc tgactcccca gatcctagtt ctctcctggg aatactcctg gatattattt
     1861 cttatgttgg ggtgggcttt tccatcttga gcttggcagc ctgtctagtt gtggaagctg
     1921 tggtgtggaa atcggtgacc aagaatcgga cttcttatat gcgccacacc tgcatagtga
     1981 atatcgctgc ctcccttctg gtcgccaaca cctggttcat tgtggtcgct gccatccagg
     2041 acaatcgcta catactctgc aagacagcct gtgtggctgc caccttcttc atccacttct
     2101 tctacctcag cgtcttcttc tggatgctga cactgggcct catgctgttc tatcgcctgg
     2161 ttttcattct gcatgaaaca agcaggtcca ctcagaaagc cattgccttc tgtcttggct
     2221 atggctgccc acttgccatc tcggtcatca cgctgggagc cacccagccc cgggaagtct
     2281 atacgaggaa gaatgtctgt tggctcaact gggaggacac caaggccctg ctggctttcg
     2341 ccatcccagc actgatcatt gtggtggtga acataaccat cactattgtg gtcatcacca
     2401 agatcctgag gccttccatt ggagacaagc catgcaagca ggagaagagc agcctgtttc
     2461 agatcagcaa gagcattggg gtcctcacac cactcttggg cctcacttgg ggttttggtc
     2521 tcaccactgt gttcccaggg accaaccttg tgttccatat catatttgcc atcctcaatg
     2581 tcttccaggg attattcatt ttactctttg gatgcctctg ggatctgaag gtacaggaag
     2641 ctttgctgaa taagttttca ttgtcgagat ggtcttcaca gcactcaaag tcaacatccc
     2701 tgggttcatc cacacctgtg ttttctatga gttctccaat atcaaggaga tttaacaatt
     2761 tgtttggtaa aacaggaacg tataatgttt ccaccccaga agcaaccagc tcatccctgg
     2821 aaaactcatc cagtgcttct tcgttgctca acggttctga ctacaaagac gatgacgaca
     2881 agtgaagatc tgacaaaact cacacatgcc caccgtgccc agcacctgaa ctcctggggg
     2941 gaccgtcagt cttcctcttc cccccaaaac ccaaggacac cctcatgatc tcccggaccc
     3001 ctgaggtcac atgcgtggtg gtggacgtga gccacgaaga ccctgaggtc aagttcaact
     3061 ggtacgtgga cggcgtggag gtgcataatg ccaagacaaa gccgcgggag gagcagtaca
     3121 acagcacgta ccgtgtggtc agcgtcctca ccgtcctgca ccaggactgg ctgaatggca
     3181 aggagtacaa gtgcaaggtc tccaacaaag ccctcccagc ccccatcgag aaaaccatct
     3241 ccaaagccaa agggcagccc cgagaaccac aggtgtacac cctgccccca tcccgggagg
     3301 agatgaccaa gaaccaggtc agcctgacct gcctggtcaa aggcttctat cccagcgaca
     3361 tcgccgtgga gtgggagagc aatgggcagc cggagaacaa ctacaagacc acgcctcccg
     3421 tgctggactc cgacggctcc ttcttcctct acagcaagct caccgtggac aagagcaggt
     3481 ggcagcaggg gaacgtcttc tcatgctccg tgatgcacga ggctctgcac aaccactaca
     3541 cgcagaagag cctctccctg tctccgggta aatgagtgct agctggccag acatgataag
     3601 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
     3661 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
     3721 caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta
     3781 aagcaagtaa aacctctaca aatgtggtat ggaattaatt ctaaaataca gcatagcaaa
     3841 actttaacct ccaaatcaag cctctacttg aatccttttc tgagggatga ataaggcata
     3901 ggcatcaggg gctgttgcca atgtgcatta gctgtttgca gcctcacctt ctttcatgga
     3961 gtttaagata tagtgtattt tcccaaggtt tgaactagct cttcatttct ttatgtttta
     4021 aatgcactga cctcccacat tcccttttta gtaaaatatt cagaaataat ttaaatacat
     4081 cattgcaatg aaaataaatg ttttttatta ggcagaatcc agatgctcaa ggcccttcat
     4141 aatatccccc agtttagtag ttggacttag ggaacaaagg aacctttaat agaaattgga
     4201 cagcaagaaa gcgagcttct agcttatcct cagtcctgct cctctgccac aaagtgcacg
     4261 cagttgccgg ccgggtcgcg cagggcgaac tcccgccccc acggctgctc gccgatctcg
     4321 gtcatggccg gcccggaggc gtcccggaag ttcgtggaca cgacctccga ccactcggcg
     4381 tacagctcgt ccaggccgcg cacccacacc caggccaggg tgttgtccgg caccacctgg
     4441 tcctggaccg cgctgatgaa cagggtcacg tcgtcccgga ccacaccggc gaagtcgtcc
     4501 tccacgaagt cccgggagaa cccgagccgg tcggtccaga actcgaccgc tccggcgacg
     4561 tcgcgcgcgg tgagcaccgg aacggcactg gtcaacttgg ccatgatggc tcctcctgtc
     4621 aggagaggaa agagaagaag gttagtacaa ttgctatagt gagttgtatt atactatgca
     4681 gatatactat gccaatgatt aattgtcaaa ctagggctgc agggttcata gtgccacttt
     4741 tcctgcactg ccccatctcc tgcccaccct ttcccaggca tagacagtca gtgacttacc
     4801 aaactcacag gagggagaag gcagaagctt gagacagacc cgcgggaccg ccgaactgcg
     4861 aggggacgtg gctagggcgg cttcttttat ggtgcgccgg ccctcggagg cagggcgctc
     4921 ggggaggcct agcggccaat ctgcggtggc aggaggcggg gccgaaggcc gtgcctgacc
     4981 aatccggagc acataggagt ctcagccccc cgccccaaag caaggggaag tcacgcgcct
     5041 gtagcgccag cgtgttgtga aatgggggct tgggggggtt ggggccctga ctagtcaaaa
     5101 caaactccca ttgacgtcaa tggggtggag acttggaaat ccccgtgagt caaaccgcta
     5161 tccacgccca ttgatgtact gccaaaaccg catcatcatg gtaatagcga tgactaatac
     5221 gtagatgtac tgccaagtag gaaagtccca taaggtcatg tactgggcat aatgccaggc
     5281 gggccattta ccgtcattga cgtcaatagg gggcgtactt ggcatatgat acacttgatg
     5341 tactgccaag tgggcagttt accgtaaata ctccacccat tgacgtcaat ggaaagtccc
     5401 tattggcgtt actatgggaa catacgtcat tattgacgtc aatgggcggg ggtcgttggg
     5461 cggtcagcca ggcgggccat ttaccgtaag ttatgtaacg cctgcaggtt aattaagaac
     5521 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     5581 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     5641 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     5701 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     5761 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     5821 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     5881 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     5941 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     6001 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     6061 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     6121 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     6181 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     6241 atggctagtt aattaacatt taaatcagcg gccgcaataa aatatcttta ttttcattac
     6301 atctgtgtgt tggttttttg tgtgaatcgt aactaacata cgctctccat caaaacaaaa
     6361 cgaaacaaaa caaactagca aaataggctg tccccagtgc aagtgcaggt gccagaacat
     6421 ttctctatcg aa
//
