LOCUS       sequence-256103-        5080 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Empty backbone for VSVG tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5080)
  AUTHORS   Smolko CM, Janes KA
  TITLE     An ultrasensitive fiveplex activity assay for cellular kinases.
  JOURNAL   Sci Rep. 2019 Dec 19;9(1):19409. doi: 10.1038/s41598-019-55998-8.
  PUBMED    31857650
REFERENCE   2  (bases 1 to 5080)
  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: "Sci Rep."; 
            date: "2019-12-19"; volume: "9(1):19409. doi"; pages: " 
            10.1038/s41598-019-55998-8"
COMMENT     Sequence Label: pGEX-4T-1-3xVSVG
FEATURES             Location/Qualifiers
     source          1..5080
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     82..99
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        172..249
                     /gene="lacI"
                     /label=lacI promoter
     CDS             250..1332
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(269..288)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(1381..1386,1387..1404,1405..1411)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1419..1435
                     /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)."
     primer_bind     1443..1459
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1443..1459
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1455..1982
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(1475..1492)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1475..1491)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1484..1506)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(2084..2089,2090..2105,2106..2112)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    2120..2136
                     /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)."
     CDS             2159..2812
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     2770..2792
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             2819..2836
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             2840..2872
                     /codon_start=1
                     /product="epitope tag from vesicular stomatitis virus G 
                     protein"
                     /label=VSV-G tag
                     /translation="YTDIEMNRLGK"
     CDS             2873..2905
                     /codon_start=1
                     /product="epitope tag from vesicular stomatitis virus G 
                     protein"
                     /label=VSV-G tag
                     /translation="YTDIEMNRLGK"
     CDS             2906..2938
                     /codon_start=1
                     /product="epitope tag from vesicular stomatitis virus G 
                     protein"
                     /label=VSV-G tag
                     /translation="YTDIEMNRLGK"
     primer_bind     complement(3031..3053)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3198..3216)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3284..3388
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3389..3457,3458..4249)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3607..3626)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4420..5008
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4909..4928
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
       61 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg
      121 cggtattttc tccttacgca tctgtgcggt atttcacacc gcataaattc cgacaccatc
      181 gaatggcgca aaacctttcg cggtatggca tgatagcgcc cggaagagag tcaattcagg
      241 gtggtgaatg tgaaaccagt aacgttatac gatgtcgcag agtatgccgg tgtctcttat
      301 cagaccgttt cccgcgtggt gaaccaggcc agccacgttt ctgcgaaaac gcgggaaaaa
      361 gtggaagcgg cgatggcgga gctgaattac attcccaacc gcgtggcaca acaactggcg
      421 ggcaaacagt cgttgctgat tggcgttgcc acctccagtc tggccctgca cgcgccgtcg
      481 caaattgtcg cggcgattaa atctcgcgcc gatcaactgg gtgccagcgt ggtggtgtcg
      541 atggtagaac gaagcggcgt cgaagcctgt aaagcggcgg tgcacaatct tctcgcgcaa
      601 cgcgtcagtg ggctgatcat taactatccg ctggatgacc aggatgccat tgctgtggaa
      661 gctgcctgca ctaatgttcc ggcgttattt cttgatgtct ctgaccagac acccatcaac
      721 agtattattt tctcccatga agacggtacg cgactgggcg tggagcatct ggtcgcattg
      781 ggtcaccagc aaatcgcgct gttagcgggc ccattaagtt ctgtctcggc gcgtctgcgt
      841 ctggctggct ggcataaata tctcactcgc aatcaaattc agccgatagc ggaacgggaa
      901 ggcgactgga gtgccatgtc cggttttcaa caaaccatgc aaatgctgaa tgagggcatc
      961 gttcccactg cgatgctggt tgccaacgat cagatggcgc tgggcgcaat gcgcgccatt
     1021 accgagtccg ggctgcgcgt tggtgcggat atctcggtag tgggatacga cgataccgaa
     1081 gacagctcat gttatatccc gccgttaacc accatcaaac aggattttcg cctgctgggg
     1141 caaaccagcg tggaccgctt gctgcaactc tctcagggcc aggcggtgaa gggcaatcag
     1201 ctgttgcccg tctcactggt gaaaagaaaa accaccctgg cgcccaatac gcaaaccgcc
     1261 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
     1321 agcgggcagt gagcgcaacg caattaatgt aagttagctc actcattagg caccccaggc
     1381 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
     1441 cacaggaaac agctatgacc atgattacgg attcactggc cgtcgtttta caacgtcgtg
     1501 actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca
     1561 gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga
     1621 atggcgaatg gcgctttgcc tggtttccgg caccagaagc ggtgccggaa agctggctgg
     1681 agtgcgatct tcctgaggcc gatactgtcg tcgtcccctc aaactggcag atgcacggtt
     1741 acgatgcgcc catctacacc aacgtgacct atcccattac ggtcaatccg ccgtttgttc
     1801 ccacggagaa tccgacgggt tgttactcgc tcacatttaa tgttgatgaa agctggctac
     1861 aggaaggcca gacgcgaatt atttttgatg gcgttggaat tagcttatcg actgcacggt
     1921 gcaccaatgc ttctggcgtc aggcagccat cggaagctgt ggtatggctg tgcaggtcgt
     1981 aaatcactgc ataattcgtg tcgctcaagg cgcactcccg ttctggataa tgttttttgc
     2041 gccgacatca taacggttct ggcaaatatt ctgaaatgag ctgttgacaa ttaatcatcg
     2101 gctcgtataa tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagtattcat
     2161 gtcccctata ctaggttatt ggaaaattaa gggccttgtg caacccactc gacttctttt
     2221 ggaatatctt gaagaaaaat atgaagagca tttgtatgag cgcgatgaag gtgataaatg
     2281 gcgaaacaaa aagtttgaat tgggtttgga gtttcccaat cttccttatt atattgatgg
     2341 tgatgttaaa ttaacacagt ctatggccat catacgttat atagctgaca agcacaacat
     2401 gttgggtggt tgtccaaaag agcgtgcaga gatttcaatg cttgaaggag cggttttgga
     2461 tattagatac ggtgtttcga gaattgcata tagtaaagac tttgaaactc tcaaagttga
     2521 ttttcttagc aagctacctg aaatgctgaa aatgttcgaa gatcgtttat gtcataaaac
     2581 atatttaaat ggtgatcatg taacccatcc tgacttcatg ttgtatgacg ctcttgatgt
     2641 tgttttatac atggacccaa tgtgcctgga tgcgttccca aaattagttt gttttaaaaa
     2701 acgtattgaa gctatcccac aaattgataa gtacttgaaa tccagcaagt atatagcatg
     2761 gcctttgcag ggctggcaag ccacgtttgg tggtggcgac catcctccaa aatcggatct
     2821 ggttccgcgt ggatctatgt acaccgatat agagatgaac aggctgggaa agtacaccga
     2881 tatagagatg aacaggctgg gaaagtacac cgatatagag atgaacaggc tgggaaagaa
     2941 gcttggatcc gaattcccgg gtcgactcga gcggccgcat cgtgactgac tgacgatctg
     3001 cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     3061 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     3121 tgttggcggg tgtcggggcg cagccatgac ccagtcacgt agcgatagcg gagtgtataa
     3181 ttcttgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     3241 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     3301 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     3361 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     3421 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     3481 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     3541 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     3601 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg
     3661 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     3721 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     3781 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     3841 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     3901 accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact
     3961 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     4021 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     4081 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     4141 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     4201 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     4261 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     4321 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     4381 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     4441 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     4501 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     4561 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     4621 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     4681 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     4741 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     4801 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     4861 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     4921 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     4981 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     5041 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
//
