LOCUS       sequence-392575-        7909 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pCDNA3.1 with Kozak (GCCACC), ADGRL3 signal peptide, SNAP-tag, GS 
            linker, and Flag-tag (EK site) ahead of ADGRL3-CTF.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7909)
  AUTHORS   Perry-Hauser NA, VanDyck MW, Lee KH, Shi L, Javitch JA
  TITLE     Disentangling autoproteolytic cleavage from tethered 
            agonist-dependent activation of the adhesion receptor ADGRL3.
  JOURNAL   J Biol Chem. 2022 Dec;298(12):102594. doi: 
            10.1016/j.jbc.2022.102594. Epub 2022 Oct 14.
  PUBMED    36244455
REFERENCE   2  (bases 1 to 7909)
  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: "J Biol 
            Chem."; date: "2022-12"; volume: "298(12):102594. doi"; pages: " 
            10.1016/j.jbc.2022.102594. Epub 2022 Oct 14"
COMMENT     Sequence Label: pCDNA3.1-Kozak-SP-SNAPf-lk-EK-ADGRL3-CTF
FEATURES             Location/Qualifiers
     source          1..7909
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(66..83)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    72..296
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      342..770
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(429..448)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     639..660
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(779..799)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        784..1113
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      964..1099
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1026..1045
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             1180..1974
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1234..1253)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1844..1863
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    2148..2269
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2185..2204)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2239..2258
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2318..2334)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2318..2334)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2331..2353)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2342..2358
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(2366..2372,2373..2390,2391..2396))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2411..2432
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2549..2566)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2720..3308)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2800..2819)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3479..4270,4271..4339))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4102..4121
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4340..4444)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4519..4538)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4710..5089
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5090..5293
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5244..5264
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5338..5357
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5338..5356
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5447..5992
                     /codon_start=1
                     /product="human O6-alkylguanine-DNA-alkyltransferase 
                     variant that forms covalent bonds with benzylguanine 
                     derivatives"
                     /label=SNAP-tag(R)
                     /note="fast-reacting variant, optimized for mammalian 
                     expression"
                     /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHRIIFLGKGTSAADAVE
                     VPAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLK
                     VVKFGEVISYSHLAALAGNPAATAAVKTALSGNPVPILIPCHRVVQGDLDVGGYEGGLA
                     VKEWLLAHEGHRLGKPGLG"
     CDS             6020..6043
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
ORIGIN
        1 ctgcagatat ccagcacagt ggcggccgct cgagtctaga gggcccgttt aaacccgctg
       61 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
      121 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
      181 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
      241 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc
      301 tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
      361 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
      421 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
      481 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
      541 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
      601 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
      661 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
      721 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg
      781 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
      841 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
      901 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
      961 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     1021 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga
     1081 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt
     1141 ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     1201 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     1261 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     1321 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta
     1381 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     1441 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     1501 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     1561 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     1621 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     1681 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc
     1741 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     1801 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     1861 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     1921 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     1981 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
     2041 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     2101 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg
     2161 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     2221 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta
     2281 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     2341 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     2401 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     2461 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     2521 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     2581 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     2641 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     2701 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     2761 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     2821 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     2881 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     2941 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     3001 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     3061 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     3121 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     3181 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     3241 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     3301 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     3361 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     3421 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     3481 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     3541 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     3601 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     3661 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     3721 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     3781 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     3841 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     3901 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     3961 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     4021 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     4081 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     4141 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     4201 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     4261 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     4321 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     4381 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     4441 cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc
     4501 cctatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct
     4561 ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag
     4621 gcaaggcttg accgacaatt gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct
     4681 tcgcgatgta cgggccagat atacgcgttg acattgatta ttgactagtt attaatagta
     4741 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
     4801 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
     4861 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
     4921 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
     4981 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
     5041 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
     5101 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
     5161 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
     5221 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
     5281 tataagcaga gctctctggc taactagaga acccactgct tactggctta tcgaaattaa
     5341 tacgactcac tatagggaga cccaagctgg ctagcgttta aacttagcca tgtggccacc
     5401 acagctactc atcctcacga tgcttttagc acctgtagtt catggtatgg acaaagactg
     5461 cgaaatgaag cgcaccaccc tggatagccc tctgggcaag ctggaactgt ctgggtgcga
     5521 acagggcctg caccgtatca tcttcctggg caaaggaaca tctgccgccg acgccgtgga
     5581 agtgcctgcc ccagccgccg tgctgggcgg accagagcca ctgatgcagg ccaccgcctg
     5641 gctcaacgcc tactttcacc agcctgaggc catcgaggag ttccctgtgc cagccctgca
     5701 ccacccagtg ttccagcagg agagctttac ccgccaggtg ctgtggaaac tgctgaaagt
     5761 ggtgaagttc ggagaggtca tcagctacag ccacctggcc gccctggccg gcaatcccgc
     5821 cgccaccgcc gccgtgaaaa ccgccctgag cggaaatccc gtgcccattc tgatcccctg
     5881 ccaccgggtg gtgcagggcg acctggacgt ggggggctac gagggcgggc tcgccgtgaa
     5941 agagtggctg ctggcccacg agggccacag actgggcaag cctgggctgg gtggcggctc
     6001 tggcggcagc ggcggaagcg attataagga cgacgacgac aagaccaact ttgcagtatt
     6061 gatggcacat gtggaagtta agcacagcga tgcagtccac gatcttcttc tggatgtgat
     6121 cacgtgggtt ggcatcttgc tgtcccttgt ctgtctcctg atttgcatct tcacattctg
     6181 cttcttccgg ggactccaga gtgaccgcaa caccattcac aagaacctct gcatcagcct
     6241 gttcgtggca gaactgctct tcctgattgg aatcaacaga accgaccaac caattgcctg
     6301 tgcagtgttc gcggctcttt tgcatttctt cttcttggcg gccttcacct ggatgtttct
     6361 agaaggggtg cagctgtata tcatgctggt ggaggtcttt gagagcgagc attcccgtag
     6421 gaagtacttt tatctggtcg gctatgggat gcccgcgctc atcgtggccg tttctgcggc
     6481 cgtcgactac aggagctacg gaacagacaa agtatgttgg cttcgccttg acacctactt
     6541 catttggagt tttataggac cggcgacctt gataattatg ctgaatgtca tcttccttgg
     6601 gattgcttta tacaaaatgt ttcaccatac tgccatactg aaacctgaat caggctgtct
     6661 tgacaatatc aactatgagg ataacagacc cttcatcaag tcatgggtta taggtgcaat
     6721 agctctgctc tgcctgttag gattgacctg ggcctttgga ctcatgtata ttaatgaaag
     6781 cacagtcatc atggcgtatc tcttcaccat tttcaattct ctacagggaa tgtttatatt
     6841 cattttccac tgtgttctac agaagaaggt acggaaagag tatgggaaat gcctccgcac
     6901 gcattgctgt agtgggaaaa gcacggagag ttcgattggc tcagggaaaa catctggttc
     6961 tcgaactcca ggacggtact ccacaggctc gcagagccgg attcggagaa tgtggaatga
     7021 cactgtccga aagcagtcag agtcgtcctt catcaccgga gacataaaca gctcagcatc
     7081 gctcaacaga gagccctaca gagagacaaa ggggcttctg aacaatgcca gggatacaag
     7141 tgtcatggat actctaccac tgaatggtaa ccatggcaac agttacagca ttgctggcgg
     7201 cgaatacctg agcaactgtg tgcaaattat agaccgtggc tataaccaca acgagaccgc
     7261 cctagagaaa aagatcctaa aggaactcac ttccaactat atcccttcat acctgaacaa
     7321 ccacgagcgc tccagcgaac agaaccggaa tatgatgaac aaactggtga acaacttagg
     7381 cagcgggagt gaagatgacg ccatcgtcct ggatgacgcc gcgtccttta accacgagga
     7441 gagtctgggc ctggaactca ttcacgagga atccgatgct cccttgctgc ccccgagggt
     7501 ttactccacg gataaccacc agccacacca ttacagcagg aggcggttcc cccaggacca
     7561 cagcgagagc ttcttccctc tgctaaccga cgagcacaca gaagacctgc agtcaccgca
     7621 cagggactct ctgtacacca gcatgccagc cctggccggc gtgcccgctg cagacagtgt
     7681 gaccaccagc acccagaccg aagccgcagc ggccaagggt ggtgacgccg aagatgttta
     7741 ctacaaaagc atgccaaacc tgggctccag aaaccacgtg cacccgctgc acgcctacta
     7801 ccagctgggg cgcggcagca gcgacggatt catagttcct cctaacaaag atggggcctc
     7861 tccggagggg acttccaaag gacccgcgca cttggtcact agtctataa
//
