LOCUS       sequence-430304-        6195 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 6195)
  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 6195)
  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-ADGRL4 CTFdeltaTA-FLAG
FEATURES             Location/Qualifiers
     source          1..6195
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    452..573
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(463..482)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     517..536
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     polyA_signal    677..1071
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     CDS             complement(1085..1459)
                     /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        2052..2355
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(2435..3023)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2515..2534)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3131..3179
                     /note="synthetic polyadenylation signal"
     misc_feature    3190..3281
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     3230..3249
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        3318..3529
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             3542..3810
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     sig_peptide     3860..3919
                     /codon_start=1
                     /product="human interleukin-2 signal sequence"
                     /label=IL-2 signal sequence
                     /translation="MYRMQLLSCIALSLALVTNS"
     CDS             3920..3946
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             5054..5074
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             5909..5932
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
ORIGIN
        1 cctcaccgtc ctgcaccagg actggctgaa tggcaaggag tacaagtgca aggtctccaa
       61 caaagccctc ccagccccca tcgagaaaac catctccaaa gccaaagggc agccccgaga
      121 accacaggtg tacaccctgc ccccatcccg ggaggagatg accaagaacc aggtcagcct
      181 gacctgcctg gtcaaaggct tctatcccag cgacatcgcc gtggagtggg agagcaatgg
      241 gcagccggag aacaactaca agaccacgcc tcccgtgctg gactccgacg gctccttctt
      301 cctctacagc aagctcaccg tggacaagag caggtggcag caggggaacg tcttctcatg
      361 ctccgtgatg cacgaggctc tgcacaacca ctacacgcag aagagcctct ccctgtctcc
      421 gggtaaatga gtgctagctg gccagacatg ataagataca ttgatgagtt tggacaaacc
      481 acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta
      541 tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg
      601 tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt
      661 ggtatggaat taattctaaa atacagcata gcaaaacttt aacctccaaa tcaagcctct
      721 acttgaatcc ttttctgagg gatgaataag gcataggcat caggggctgt tgccaatgtg
      781 cattagctgt ttgcagcctc accttctttc atggagttta agatatagtg tattttccca
      841 aggtttgaac tagctcttca tttctttatg ttttaaatgc actgacctcc cacattccct
      901 ttttagtaaa atattcagaa ataatttaaa tacatcattg caatgaaaat aaatgttttt
      961 tattaggcag aatccagatg ctcaaggccc ttcataatat cccccagttt agtagttgga
     1021 cttagggaac aaaggaacct ttaatagaaa ttggacagca agaaagcgag cttctagctt
     1081 atcctcagtc ctgctcctct gccacaaagt gcacgcagtt gccggccggg tcgcgcaggg
     1141 cgaactcccg cccccacggc tgctcgccga tctcggtcat ggccggcccg gaggcgtccc
     1201 ggaagttcgt ggacacgacc tccgaccact cggcgtacag ctcgtccagg ccgcgcaccc
     1261 acacccaggc cagggtgttg tccggcacca cctggtcctg gaccgcgctg atgaacaggg
     1321 tcacgtcgtc ccggaccaca ccggcgaagt cgtcctccac gaagtcccgg gagaacccga
     1381 gccggtcggt ccagaactcg accgctccgg cgacgtcgcg cgcggtgagc accggaacgg
     1441 cactggtcaa cttggccatg atggctcctc ctgtcaggag aggaaagaga agaaggttag
     1501 tacaattgct atagtgagtt gtattatact atgcagatat actatgccaa tgattaattg
     1561 tcaaactagg gctgcagggt tcatagtgcc acttttcctg cactgcccca tctcctgccc
     1621 accctttccc aggcatagac agtcagtgac ttaccaaact cacaggaggg agaaggcaga
     1681 agcttgagac agacccgcgg gaccgccgaa ctgcgagggg acgtggctag ggcggcttct
     1741 tttatggtgc gccggccctc ggaggcaggg cgctcgggga ggcctagcgg ccaatctgcg
     1801 gtggcaggag gcggggccga aggccgtgcc tgaccaatcc ggagcacata ggagtctcag
     1861 ccccccgccc caaagcaagg ggaagtcacg cgcctgtagc gccagcgtgt tgtgaaatgg
     1921 gggcttgggg gggttggggc cctgactagt caaaacaaac tcccattgac gtcaatgggg
     1981 tggagacttg gaaatccccg tgagtcaaac cgctatccac gcccattgat gtactgccaa
     2041 aaccgcatca tcatggtaat agcgatgact aatacgtaga tgtactgcca agtaggaaag
     2101 tcccataagg tcatgtactg ggcataatgc caggcgggcc atttaccgtc attgacgtca
     2161 atagggggcg tacttggcat atgatacact tgatgtactg ccaagtgggc agtttaccgt
     2221 aaatactcca cccattgacg tcaatggaaa gtccctattg gcgttactat gggaacatac
     2281 gtcattattg acgtcaatgg gcgggggtcg ttgggcggtc agccaggcgg gccatttacc
     2341 gtaagttatg taacgcctgc aggttaatta agaacatgtg agcaaaaggc cagcaaaagg
     2401 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     2461 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     2521 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     2581 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     2641 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     2701 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     2761 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     2821 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     2881 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     2941 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     3001 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     3061 agtggaacga aaactcacgt taagggattt tggtcatggc tagttaatta acatttaaat
     3121 cagcggccgc aataaaatat ctttattttc attacatctg tgtgttggtt ttttgtgtga
     3181 atcgtaacta acatacgctc tccatcaaaa caaaacgaaa caaaacaaac tagcaaaata
     3241 ggctgtcccc agtgcaagtg caggtgccag aacatttctc tatcgaagga tctgcgatcg
     3301 ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg
     3361 aggggtcggc aattgaacgg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga
     3421 tgtcgtgtac tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt
     3481 agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc agaacacagc tgaagcttcg
     3541 aggggctcgc atctctcctt cacgcgcccg ccgccctacc tgaggccgcc atccacgccg
     3601 gttgagtcgc gttctgccgc ctcccgcctg tggtgcctcc tgaactgcgt ccgccgtcta
     3661 ggtaagttta aagctcaggt cgagaccggg cctttgtccg gcgctccctt ggagcctacc
     3721 tagactcagc cggctctcca cgctttgcct gaccctgctt gctcaactct acgtctttgt
     3781 ttcgttttct gttctgcgcc gttacagatc caagctgtga ccggcgccta cctgagatca
     3841 ccggcgaagg agggccacca tgtacaggat gcaactcctg tcttgcattg cactaagtct
     3901 tgcacttgtc acgaattcgt acccatacga tgttccagat tacgctggat ccaaaattga
     3961 agaaggcaaa ctggtgattt ggattaacgg cgataaaggc tataacggcc tggcggaagt
     4021 gggcaaaaaa tttgaaaaag ataccggcat taaagtgacc gtggaacatc cggataaact
     4081 ggaagaaaaa tttccgcagg tggcggcgac cggcgatggc ccggatatta ttttttgggc
     4141 gcatgatcgc tttggcggct atgcgcagag cggcctgctg gcggaaatta ccccggataa
     4201 agcgtttcag gataaactgt atccgtttac ctgggatgcg gtgcgctata acggcaaact
     4261 gattgcgtat ccgattgcgg tggaagcgct gagcctgatt tataacaaag atctgctgcc
     4321 gaacccgccg aaaacctggg aagaaattcc ggcgctggat aaagaactga aagcgaaagg
     4381 caaaagcgcg ctgatgttta acctgcagga accgtatttt acctggccgc tgattgcggc
     4441 ggatggcggc tatgcgttta aatatgaaaa cggcaaatat gatattaaag atgtgggcgt
     4501 ggataacgcg ggcgcgaaag cgggcctgac ctttctggtg gatctgatta aaaacaaaca
     4561 tatgaacgcg gataccgatt atagcattgc ggaagcggcg tttaacaaag gcgaaaccgc
     4621 gatgaccatt aacggcccgt gggcgtggag caacattgat accagcaaag tgaactatgg
     4681 cgtgaccgtg ctgccgacct ttaaaggcca gccgagcaaa ccgtttgtgg gcgtgctgag
     4741 cgcgggcatt aacgcggcga gcccgaacaa agaactggcg aaagaatttc tggaaaacta
     4801 tctgctgacc gatgaaggcc tggaagcggt gaacaaagat aaaccgctgg gcgcggtggc
     4861 gctgaaaagc tatgaagaag aactggcgaa agatccgcgc attgcggcga ccatggaaaa
     4921 cgcgcagaaa ggcgaaatta tgccgaacat tccgcagatg agcgcgtttt ggtatgcggt
     4981 gcgcaccgcg gtgattaacg cggcgagcgg ccgccagacc gtggatgaag cgctgaaaga
     5041 tgcgcagacc aacgagaacc tttacttcca aagctccggg cctagcatcg gcatcaagga
     5101 ctataatatc ttgaccagga ttacacagct gggtatcatt atctctttga tctgtttggc
     5161 tatttgtatc ttcaccttct ggtttttcag tgaaatccag tctacccgga caaccatcca
     5221 taagaatctt tgttgctccc tgttcctggc cgagctggtg ttcctggtgg ggatcaatac
     5281 taacactaac aagctgttct gttcaattat cgccggcttg ctgcactact ttttcctcgc
     5341 agctttcgcc tggatgtgca ttgaaggtat tcacctctat ctgattgttg tgggggtgat
     5401 ttacaacaaa ggcttcctgc acaagaactt ctacatcttt ggctacctgt ctcccgccgt
     5461 cgtggttgga tttagcgcgg ccctggggta tcgttattac ggcacaacca aggtttgttg
     5521 gttgagcaca gaaaataact tcatctggtc tttcattggt cctgcatgcc tgattatcct
     5581 ggtcaacctg cttgcctttg gcgtaattat ctacaaggtt ttccgccaca ccgcgggcct
     5641 gaagcccgaa gtgtcttgtt tcgaaaacat ccggtcctgc gctcgcggag ctctggccct
     5701 cctgtttctc ctgggcacca cttggatttt cggcgtcctg cacgtcgtgc acgcctccgt
     5761 ggtcacagcc tacctgttca ccgtgtctaa cgcattccag ggcatgttca tcttcctgtt
     5821 cctgtgtgta ctgtcccgca agatccaaga agagtattac cgtctgttca agaacgtgcc
     5881 atgctgtttt ggctgtctca gaggttctga ctacaaagac gatgacgaca agtgaagatc
     5941 tgacaaaact cacacatgcc caccgtgccc agcacctgaa ctcctggggg gaccgtcagt
     6001 cttcctcttc cccccaaaac ccaaggacac cctcatgatc tcccggaccc ctgaggtcac
     6061 atgcgtggtg gtggacgtga gccacgaaga ccctgaggtc aagttcaact ggtacgtgga
     6121 cggcgtggag gtgcataatg ccaagacaaa gccgcgggag gagcagtaca acagcacgta
     6181 ccgtgtggtc agcgt
//
