LOCUS       sequence-430003-        6276 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 6276)
  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 6276)
  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-ADGRF4 CTFdeltaTA-FLAG
FEATURES             Location/Qualifiers
     source          1..6276
                     /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             1767..1787
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             2703..2726
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    3441..3562
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3452..3471)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3506..3525
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     polyA_signal    3666..4060
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     CDS             complement(4074..4448)
                     /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        5041..5344
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(5424..6012)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5504..5523)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    6120..6168
                     /note="synthetic polyadenylation signal"
     misc_feature    6179..6270
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     6219..6238
                     /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 ccaatcttcc aagtccatga
     1801 ccgataaggt gctggattac atcacctgca ttggtctgtc cgtgtccatc ctgagcctgg
     1861 tgctctgttt gattatcgaa gccaccgtgt ggagccgggt cgtagtgact gagatttcct
     1921 acatgcggca tgtgtgcatc gtgaacattg ccgtgtcttt gctgaccgca aatgtgtggt
     1981 tcattatcgg ttctcatttt aacatcaagg cccaggatta caatatgtgc gtggcagtga
     2041 cctttttctc ccattttttc tatctgtctc tgtttttctg gatgctgttc aaggcgcttc
     2101 tgattatcta cggaatcctg gtcattttca ggcgcatgat gaagagtcgc atgatggtca
     2161 tcggctttgc tatcggatac ggctgtccac tgattatcgc cgtgacaacc gttgccatca
     2221 ccgaacccga aaagggatac atgcgtcctg aggcctgctg gctgaactgg gacaacacca
     2281 aagccttgct ggctttcgcc atccctgcct ttgttatcgt tgcggtgaac ttgattgtgg
     2341 tcctggtagt ggccgtgaac actcagcgcc cctccattgg aagctccaag agccaggacg
     2401 ttgtgatcat tatgaggatc agcaagaatg tcgcgatcct gaccccgctg cttgggctta
     2461 cctggggttt cggcatcgct accctgattg agggcaccag cttgaccttc cacattatct
     2521 ttgccctcct gaatgctttc caaggatttt tcattctgct tttcggaacg atcatggacc
     2581 acaagatccg cgacgctctg cgcatgagaa tgtctagcct gaaggggaaa tctagagcag
     2641 ctgaaaacgc atctctgggc cccactaacg gctcaaagct gatgaatcgc cagggaggtt
     2701 ctgactacaa agacgatgac gacaagtgaa gatctgacaa aactcacaca tgcccaccgt
     2761 gcccagcacc tgaactcctg gggggaccgt cagtcttcct cttcccccca aaacccaagg
     2821 acaccctcat gatctcccgg acccctgagg tcacatgcgt ggtggtggac gtgagccacg
     2881 aagaccctga ggtcaagttc aactggtacg tggacggcgt ggaggtgcat aatgccaaga
     2941 caaagccgcg ggaggagcag tacaacagca cgtaccgtgt ggtcagcgtc ctcaccgtcc
     3001 tgcaccagga ctggctgaat ggcaaggagt acaagtgcaa ggtctccaac aaagccctcc
     3061 cagcccccat cgagaaaacc atctccaaag ccaaagggca gccccgagaa ccacaggtgt
     3121 acaccctgcc cccatcccgg gaggagatga ccaagaacca ggtcagcctg acctgcctgg
     3181 tcaaaggctt ctatcccagc gacatcgccg tggagtggga gagcaatggg cagccggaga
     3241 acaactacaa gaccacgcct cccgtgctgg actccgacgg ctccttcttc ctctacagca
     3301 agctcaccgt ggacaagagc aggtggcagc aggggaacgt cttctcatgc tccgtgatgc
     3361 acgaggctct gcacaaccac tacacgcaga agagcctctc cctgtctccg ggtaaatgag
     3421 tgctagctgg ccagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     3481 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     3541 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     3601 gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtatggaatt
     3661 aattctaaaa tacagcatag caaaacttta acctccaaat caagcctcta cttgaatcct
     3721 tttctgaggg atgaataagg cataggcatc aggggctgtt gccaatgtgc attagctgtt
     3781 tgcagcctca ccttctttca tggagtttaa gatatagtgt attttcccaa ggtttgaact
     3841 agctcttcat ttctttatgt tttaaatgca ctgacctccc acattccctt tttagtaaaa
     3901 tattcagaaa taatttaaat acatcattgc aatgaaaata aatgtttttt attaggcaga
     3961 atccagatgc tcaaggccct tcataatatc ccccagttta gtagttggac ttagggaaca
     4021 aaggaacctt taatagaaat tggacagcaa gaaagcgagc ttctagctta tcctcagtcc
     4081 tgctcctctg ccacaaagtg cacgcagttg ccggccgggt cgcgcagggc gaactcccgc
     4141 ccccacggct gctcgccgat ctcggtcatg gccggcccgg aggcgtcccg gaagttcgtg
     4201 gacacgacct ccgaccactc ggcgtacagc tcgtccaggc cgcgcaccca cacccaggcc
     4261 agggtgttgt ccggcaccac ctggtcctgg accgcgctga tgaacagggt cacgtcgtcc
     4321 cggaccacac cggcgaagtc gtcctccacg aagtcccggg agaacccgag ccggtcggtc
     4381 cagaactcga ccgctccggc gacgtcgcgc gcggtgagca ccggaacggc actggtcaac
     4441 ttggccatga tggctcctcc tgtcaggaga ggaaagagaa gaaggttagt acaattgcta
     4501 tagtgagttg tattatacta tgcagatata ctatgccaat gattaattgt caaactaggg
     4561 ctgcagggtt catagtgcca cttttcctgc actgccccat ctcctgccca ccctttccca
     4621 ggcatagaca gtcagtgact taccaaactc acaggaggga gaaggcagaa gcttgagaca
     4681 gacccgcggg accgccgaac tgcgagggga cgtggctagg gcggcttctt ttatggtgcg
     4741 ccggccctcg gaggcagggc gctcggggag gcctagcggc caatctgcgg tggcaggagg
     4801 cggggccgaa ggccgtgcct gaccaatccg gagcacatag gagtctcagc cccccgcccc
     4861 aaagcaaggg gaagtcacgc gcctgtagcg ccagcgtgtt gtgaaatggg ggcttggggg
     4921 ggttggggcc ctgactagtc aaaacaaact cccattgacg tcaatggggt ggagacttgg
     4981 aaatccccgt gagtcaaacc gctatccacg cccattgatg tactgccaaa accgcatcat
     5041 catggtaata gcgatgacta atacgtagat gtactgccaa gtaggaaagt cccataaggt
     5101 catgtactgg gcataatgcc aggcgggcca tttaccgtca ttgacgtcaa tagggggcgt
     5161 acttggcata tgatacactt gatgtactgc caagtgggca gtttaccgta aatactccac
     5221 ccattgacgt caatggaaag tccctattgg cgttactatg ggaacatacg tcattattga
     5281 cgtcaatggg cgggggtcgt tgggcggtca gccaggcggg ccatttaccg taagttatgt
     5341 aacgcctgca ggttaattaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     5401 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     5461 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     5521 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     5581 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     5641 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     5701 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     5761 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     5821 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     5881 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     5941 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     6001 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     6061 aactcacgtt aagggatttt ggtcatggct agttaattaa catttaaatc agcggccgca
     6121 ataaaatatc tttattttca ttacatctgt gtgttggttt tttgtgtgaa tcgtaactaa
     6181 catacgctct ccatcaaaac aaaacgaaac aaaacaaact agcaaaatag gctgtcccca
     6241 gtgcaagtgc aggtgccaga acatttctct atcgaa
//
