LOCUS       sequence-430285-        6345 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 6345)
  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 6345)
  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-ADGRF3 CTF-FLAG
FEATURES             Location/Qualifiers
     source          1..6345
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             450..824
                     /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"
     polyA_signal    838..1232
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     polyA_signal    1336..1457
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1373..1392)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1427..1446
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(2172..2195)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(4308..4334)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     sig_peptide     complement(4335..4394)
                     /codon_start=1
                     /product="human interleukin-2 signal sequence"
                     /label=IL-2 signal sequence
                     /translation="MYRMQLLSCIALSLALVTNS"
     LTR             4444..4712
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     promoter        complement(4725..4936)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     misc_feature    4973..5064
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     complement(5005..5024)
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     polyA_signal    5075..5123
                     /note="synthetic polyadenylation signal"
     rep_origin      5231..5819
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5720..5739
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        5899..6202
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 acgctggcgc tacaggcgcg tgacttcccc ttgctttggg gcggggggct gagactccta
       61 tgtgctccgg attggtcagg cacggccttc ggccccgcct cctgccaccg cagattggcc
      121 gctaggcctc cccgagcgcc ctgcctccga gggccggcgc accataaaag aagccgccct
      181 agccacgtcc cctcgcagtt cggcggtccc gcgggtctgt ctcaagcttc tgccttctcc
      241 ctcctgtgag tttggtaagt cactgactgt ctatgcctgg gaaagggtgg gcaggagatg
      301 gggcagtgca ggaaaagtgg cactatgaac cctgcagccc tagtttgaca attaatcatt
      361 ggcatagtat atctgcatag tataatacaa ctcactatag caattgtact aaccttcttc
      421 tctttcctct cctgacagga ggagccatca tggccaagtt gaccagtgcc gttccggtgc
      481 tcaccgcgcg cgacgtcgcc ggagcggtcg agttctggac cgaccggctc gggttctccc
      541 gggacttcgt ggaggacgac ttcgccggtg tggtccggga cgacgtgacc ctgttcatca
      601 gcgcggtcca ggaccaggtg gtgccggaca acaccctggc ctgggtgtgg gtgcgcggcc
      661 tggacgagct gtacgccgag tggtcggagg tcgtgtccac gaacttccgg gacgcctccg
      721 ggccggccat gaccgagatc ggcgagcagc cgtgggggcg ggagttcgcc ctgcgcgacc
      781 cggccggcaa ctgcgtgcac tttgtggcag aggagcagga ctgaggataa gctagaagct
      841 cgctttcttg ctgtccaatt tctattaaag gttcctttgt tccctaagtc caactactaa
      901 actgggggat attatgaagg gccttgagca tctggattct gcctaataaa aaacatttat
      961 tttcattgca atgatgtatt taaattattt ctgaatattt tactaaaaag ggaatgtggg
     1021 aggtcagtgc atttaaaaca taaagaaatg aagagctagt tcaaaccttg ggaaaataca
     1081 ctatatctta aactccatga aagaaggtga ggctgcaaac agctaatgca cattggcaac
     1141 agcccctgat gcctatgcct tattcatccc tcagaaaagg attcaagtag aggcttgatt
     1201 tggaggttaa agttttgcta tgctgtattt tagaattaat tccataccac atttgtagag
     1261 gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat
     1321 gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     1381 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     1441 ctcatcaatg tatcttatca tgtctggcca gctagcactc atttacccgg agacagggag
     1501 aggctcttct gcgtgtagtg gttgtgcaga gcctcgtgca tcacggagca tgagaagacg
     1561 ttcccctgct gccacctgct cttgtccacg gtgagcttgc tgtagaggaa gaaggagccg
     1621 tcggagtcca gcacgggagg cgtggtcttg tagttgttct ccggctgccc attgctctcc
     1681 cactccacgg cgatgtcgct gggatagaag cctttgacca ggcaggtcag gctgacctgg
     1741 ttcttggtca tctcctcccg ggatgggggc agggtgtaca cctgtggttc tcggggctgc
     1801 cctttggctt tggagatggt tttctcgatg ggggctggga gggctttgtt ggagaccttg
     1861 cacttgtact ccttgccatt cagccagtcc tggtgcagga cggtgaggac gctgaccaca
     1921 cggtacgtgc tgttgtactg ctcctcccgc ggctttgtct tggcattatg cacctccacg
     1981 ccgtccacgt accagttgaa cttgacctca gggtcttcgt ggctcacgtc caccaccacg
     2041 catgtgacct caggggtccg ggagatcatg agggtgtcct tgggttttgg ggggaagagg
     2101 aagactgacg gtccccccag gagttcaggt gctgggcacg gtgggcatgt gtgagttttg
     2161 tcagatcttc acttgtcgtc atcgtctttg tagtcagaac cactgcgggc ggtacccata
     2221 tcttcagagg atggccatgg gataccctca gacatagatt tgcttgcaca gctcaggatc
     2281 tgcaggcaac aactcacgag actgatagta gagctcggtg cctgggcgcg gcaaaagcgc
     2341 ttacgcaacg cttcctggat tttccggtcc atcaggcagc cgaaaagcag gataaagacg
     2401 ccctgcagtg tattcaggat ggtgaagata tagtggggca cggtactaac ctcttcgagc
     2461 agggtcgcaa gacccaggcc ccaggtcaac ccgaagattg gggtgaggat aaggagggcc
     2521 ttgatgacgc ccaggagggc ctggcgcttt tctgctggcg gaccctcgga aagagaaggc
     2581 ctcagcaatt tgagcattgc catggccaag accagcccgt tgactccgat aatcgccaga
     2641 acggggccca caaaggtgta cagtgctccg cccttaccgt ccagccagca ctcgccctcg
     2701 cgcagatact ggccctgagg gaggtacagg cccagtgtaa cgccagccaa gccaaggggg
     2761 cacaggtatc ccaggagcac catcaggggc agcactctgt gttttgcgag ctggtgaaac
     2821 acgaacagaa gctggtgagc cagcaccaga gcctgggcaa gcatccagaa aaatgttgcc
     2881 aggtacagaa agtggcacag aaaggcagcg gccaggcaca gaggagagcg gggcccgggg
     2941 ctcaggaagg gggcgcccag gaaacaggtg tcggcagcca ggaggcaaaa caccatgttc
     3001 aacagcgcgg cgtggcggaa ataagaaatt ttgttgcgca cgacaacccg ccacaccagc
     3061 cagtaaactc ccaggcagac cagaagtgcc aagatgctgg cgcccagtcc aacctgagtg
     3121 agcagcgcaa gcgcgggttc ctccggcacg gtgtgggggc tcatcaaaac agagaaagcg
     3181 gtttggaagt aaaggttctc gttggtctgc gcatctttca gcgcttcatc cacggtctgg
     3241 cggccgctcg ccgcgttaat caccgcggtg cgcaccgcat accaaaacgc gctcatctgc
     3301 ggaatgttcg gcataatttc gcctttctgc gcgttttcca tggtcgccgc aatgcgcgga
     3361 tctttcgcca gttcttcttc atagcttttc agcgccaccg cgcccagcgg tttatctttg
     3421 ttcaccgctt ccaggccttc atcggtcagc agatagtttt ccagaaattc tttcgccagt
     3481 tctttgttcg ggctcgccgc gttaatgccc gcgctcagca cgcccacaaa cggtttgctc
     3541 ggctggcctt taaaggtcgg cagcacggtc acgccatagt tcactttgct ggtatcaatg
     3601 ttgctccacg cccacgggcc gttaatggtc atcgcggttt cgcctttgtt aaacgccgct
     3661 tccgcaatgc tataatcggt atccgcgttc atatgtttgt ttttaatcag atccaccaga
     3721 aaggtcaggc ccgctttcgc gcccgcgtta tccacgccca catctttaat atcatatttg
     3781 ccgttttcat atttaaacgc atagccgcca tccgccgcaa tcagcggcca ggtaaaatac
     3841 ggttcctgca ggttaaacat cagcgcgctt ttgcctttcg ctttcagttc tttatccagc
     3901 gccggaattt cttcccaggt tttcggcggg ttcggcagca gatctttgtt ataaatcagg
     3961 ctcagcgctt ccaccgcaat cggatacgca atcagtttgc cgttatagcg caccgcatcc
     4021 caggtaaacg gatacagttt atcctgaaac gctttatccg gggtaatttc cgccagcagg
     4081 ccgctctgcg catagccgcc aaagcgatca tgcgcccaaa aaataatatc cgggccatcg
     4141 ccggtcgccg ccacctgcgg aaatttttct tccagtttat ccggatgttc cacggtcact
     4201 ttaatgccgg tatctttttc aaattttttg cccacttccg ccaggccgtt atagccttta
     4261 tcgccgttaa tccaaatcac cagtttgcct tcttcaattt tggatccagc gtaatctgga
     4321 acatcgtatg ggtacgaatt cgtgacaagt gcaagactta gtgcaatgca agacaggagt
     4381 tgcatcctgt acatggtggc cctccttcgc cggtgatctc aggtaggcgc cggtcacagc
     4441 ttggatctgt aacggcgcag aacagaaaac gaaacaaaga cgtagagttg agcaagcagg
     4501 gtcaggcaaa gcgtggagag ccggctgagt ctaggtaggc tccaagggag cgccggacaa
     4561 aggcccggtc tcgacctgag ctttaaactt acctagacgg cggacgcagt tcaggaggca
     4621 ccacaggcgg gaggcggcag aacgcgactc aaccggcgtg gatggcggcc tcaggtaggg
     4681 cggcgggcgc gtgaaggaga gatgcgagcc cctcgaagct tcagctgtgt tctggcggca
     4741 aacccgttgc gaaaaagaac gttcacggcg actactgcac ttatatacgg ttctccccca
     4801 ccctcgggaa aaaggcggag ccagtacacg acatcacttt cccagtttac cccgcgccac
     4861 cttctctagg cacccgttca attgccgacc cctcccccca acttctcggg gactgtgggc
     4921 gatgtgcgct ctgcccactg acgggcaccg gagcgatcgc agatccttcg atagagaaat
     4981 gttctggcac ctgcacttgc actggggaca gcctattttg ctagtttgtt ttgtttcgtt
     5041 ttgttttgat ggagagcgta tgttagttac gattcacaca aaaaaccaac acacagatgt
     5101 aatgaaaata aagatatttt attgcggccg ctgatttaaa tgttaattaa ctagccatga
     5161 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     5221 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     5281 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     5341 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     5401 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     5461 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     5521 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     5581 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     5641 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     5701 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     5761 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     5821 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     5881 tcttaattaa cctgcaggcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
     5941 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
     6001 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
     6061 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
     6121 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
     6181 attagtcatc gctattacca tgatgatgcg gttttggcag tacatcaatg ggcgtggata
     6241 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
     6301 ttgactagtc agggccccaa cccccccaag cccccatttc acaac
//
