LOCUS       sequence-233666-        5014 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Allows cloning of gene of interest into bacterial expression vector 
            with PreScission Protease cleavable N-terminal GST tag, and a 
            retained N-terminal Strep-II tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5014)
  TITLE     Bacterial expression plasmids for production of recombinant proteins
            with a PreScission Protease cleavable GST tag and additional small 
            affinity tags
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5014)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pGEX6P1-N-Strep2
FEATURES             Location/Qualifiers
     source          1..5014
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             23..676
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     634..656
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             683..706
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             716..739
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(829..851)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(996..1014)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1082..1186
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1187..1255,1256..2047)
                     /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(1405..1424)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2218..2806
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2707..2726
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2960..2977
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        3050..3127
                     /gene="lacI"
                     /label=lacI promoter
     CDS             3128..4210
                     /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(3147..3166)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(4259..4264,4265..4282,4283..4289)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4297..4313
                     /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     4321..4337
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4321..4337
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4333..4860
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(4353..4370)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4353..4369)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4362..4384)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(4962..4967,4968..4983,4984..4990)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    4998..5014
                     /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)."
ORIGIN
        1 tttcacacag gaaacagtat tcatgtcccc tatactaggt tattggaaaa ttaagggcct
       61 tgtgcaaccc actcgacttc ttttggaata tcttgaagaa aaatatgaag agcatttgta
      121 tgagcgcgat gaaggtgata aatggcgaaa caaaaagttt gaattgggtt tggagtttcc
      181 caatcttcct tattatattg atggtgatgt taaattaaca cagtctatgg ccatcatacg
      241 ttatatagct gacaagcaca acatgttggg tggttgtcca aaagagcgtg cagagatttc
      301 aatgcttgaa ggagcggttt tggatattag atacggtgtt tcgagaattg catatagtaa
      361 agactttgaa actctcaaag ttgattttct tagcaagcta cctgaaatgc tgaaaatgtt
      421 cgaagatcgt ttatgtcata aaacatattt aaatggtgat catgtaaccc atcctgactt
      481 catgttgtat gacgctcttg atgttgtttt atacatggac ccaatgtgcc tggatgcgtt
      541 cccaaaatta gtttgtttta aaaaacgtat tgaagctatc ccacaaattg ataagtactt
      601 gaaatccagc aagtatatag catggccttt gcagggctgg caagccacgt ttggtggtgg
      661 cgaccatcct ccaaaatcgg atctggaagt tctgttccag gggcccctgg gatcctggag
      721 ccacccgcag ttcgaaaagc gatccccgga attcccgggt cgactcgagc ggccgcatcg
      781 tgactgactg acgatctgcc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
      841 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
      901 tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc agtcacgtag
      961 cgatagcgga gtgtataatt cttgaagacg aaagggcctc gtgatacgcc tatttttata
     1021 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     1081 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     1141 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     1201 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     1261 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     1321 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     1381 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg
     1441 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     1501 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     1561 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     1621 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     1681 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg cagcaatggc
     1741 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     1801 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     1861 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     1921 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     1981 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     2041 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     2101 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     2161 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     2221 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     2281 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     2341 cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa
     2401 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     2461 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     2521 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     2581 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     2641 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     2701 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     2761 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     2821 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     2881 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     2941 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg
     3001 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc ataaattccg acaccatcga
     3061 atggcgcaaa acctttcgcg gtatggcatg atagcgcccg gaagagagtc aattcagggt
     3121 ggtgaatgtg aaaccagtaa cgttatacga tgtcgcagag tatgccggtg tctcttatca
     3181 gaccgtttcc cgcgtggtga accaggccag ccacgtttct gcgaaaacgc gggaaaaagt
     3241 ggaagcggcg atggcggagc tgaattacat tcccaaccgc gtggcacaac aactggcggg
     3301 caaacagtcg ttgctgattg gcgttgccac ctccagtctg gccctgcacg cgccgtcgca
     3361 aattgtcgcg gcgattaaat ctcgcgccga tcaactgggt gccagcgtgg tggtgtcgat
     3421 ggtagaacga agcggcgtcg aagcctgtaa agcggcggtg cacaatcttc tcgcgcaacg
     3481 cgtcagtggg ctgatcatta actatccgct ggatgaccag gatgccattg ctgtggaagc
     3541 tgcctgcact aatgttccgg cgttatttct tgatgtctct gaccagacac ccatcaacag
     3601 tattattttc tcccatgaag acggtacgcg actgggcgtg gagcatctgg tcgcattggg
     3661 tcaccagcaa atcgcgctgt tagcgggccc attaagttct gtctcggcgc gtctgcgtct
     3721 ggctggctgg cataaatatc tcactcgcaa tcaaattcag ccgatagcgg aacgggaagg
     3781 cgactggagt gccatgtccg gttttcaaca aaccatgcaa atgctgaatg agggcatcgt
     3841 tcccactgcg atgctggttg ccaacgatca gatggcgctg ggcgcaatgc gcgccattac
     3901 cgagtccggg ctgcgcgttg gtgcggatat ctcggtagtg ggatacgacg ataccgaaga
     3961 cagctcatgt tatatcccgc cgttaaccac catcaaacag gattttcgcc tgctggggca
     4021 aaccagcgtg gaccgcttgc tgcaactctc tcagggccag gcggtgaagg gcaatcagct
     4081 gttgcccgtc tcactggtga aaagaaaaac cacgctagcg cccaatacgc aaaccgcctc
     4141 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
     4201 cgggcagtga gcgcaacgca attaatgtaa gttagctcac tcattaggca ccccaggctt
     4261 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
     4321 caggaaacag ctatgaccat gattacggat tcactggccg tcgttttaca acgtcgtgac
     4381 tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc
     4441 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
     4501 ggcgaatggc gctttgcctg gtttccggca ccagaagcgg tgccggaaag ctggctggag
     4561 tgcgatcttc ctgaggccga tactgtcgtc gtcccctcaa actggcagat gcacggttac
     4621 gatgcgccca tctacaccaa cgtgacctat cccattacgg tcaatccgcc gtttgttccc
     4681 acggagaatc cgacgggttg ttactcgctc acatttaatg ttgatgaaag ctggctacag
     4741 gaaggccaga cgcgaattat ttttgatggc gttggaatta gcttatcgac tgcacggtgc
     4801 accaatgctt ctggcgtcag gcagccatcg gaagctgtgg tatggctgtg caggtcgtaa
     4861 atcactgcat aattcgtgtc gctcaaggcg cactcccgtt ctggataatg ttttttgcgc
     4921 cgacatcata acggttctgg caaatattct gaaatgagct gttgacaatt aatcatcggc
     4981 tcgtataatg tgtggaattg tgagcggata acaa
//
