LOCUS       sequence-355702-        6119 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Bacterial Expression of HsTNRC6C_1281-1665.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6119)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 6119)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pGEX-6P-1-HsTNRC6C_1281-1665_P
FEATURES             Location/Qualifiers
     source          1..6119
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(183..188,189..204,205..211)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    219..235
                     /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             258..911
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     869..891
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             918..941
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(2169..2191)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2336..2354)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2422..2526
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2527..2595,2596..3387)
                     /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(2745..2764)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3558..4146
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4047..4066
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4300..4317
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4390..4467
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4468..5550
                     /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(4487..4506)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5599..5604,5605..5622,5623..5629)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5637..5653
                     /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     5661..5677
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5661..5677
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(5673..6119,1..81)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(5693..5710)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5693..5709)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5702..5724)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 agcttatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc ggaagctgtg
       61 gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc gcactcccgt
      121 tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc tgaaatgagc
      181 tgttgacaat taatcatcgg ctcgtataat gtgtggaatt gtgagcggat aacaatttca
      241 cacaggaaac agtattcatg tcccctatac taggttattg gaaaattaag ggccttgtgc
      301 aacccactcg acttcttttg gaatatcttg aagaaaaata tgaagagcat ttgtatgagc
      361 gcgatgaagg tgataaatgg cgaaacaaaa agtttgaatt gggtttggag tttcccaatc
      421 ttccttatta tattgatggt gatgttaaat taacacagtc tatggccatc atacgttata
      481 tagctgacaa gcacaacatg ttgggtggtt gtccaaaaga gcgtgcagag atttcaatgc
      541 ttgaaggagc ggttttggat attagatacg gtgtttcgag aattgcatat agtaaagact
      601 ttgaaactct caaagttgat tttcttagca agctacctga aatgctgaaa atgttcgaag
      661 atcgtttatg tcataaaaca tatttaaatg gtgatcatgt aacccatcct gacttcatgt
      721 tgtatgacgc tcttgatgtt gttttataca tggacccaat gtgcctggat gcgttcccaa
      781 aattagtttg ttttaaaaaa cgtattgaag ctatcccaca aattgataag tacttgaaat
      841 ccagcaagta tatagcatgg cctttgcagg gctggcaagc cacgtttggt ggtggcgacc
      901 atcctccaaa atcggatctg gaagttctgt tccaggggcc cctgggatcc atgaccggtg
      961 gcttgtcggt gaaggaccca tcccagtccc agtcacgcct cccccagtgg acgcacccca
     1021 actccatgga taacttgccc agtgccgctt cccccctgga gcagaaccct agcaagcatg
     1081 gtgctatccc tggaggtcta agcattgggc ctccaggtaa gtcctccatt gatgactcct
     1141 atggccggta cgatttaatc cagaacagtg agtcaccagc cagtcctccc gtagctgttc
     1201 cccatagctg gtcacgtgcc aaatctgaca gtgataaaat ctcaaatggc tctagcatca
     1261 actggccccc agaattccat ccgggagttc catggaaagg actgcagaat attgaccctg
     1321 agaatgaccc tgacgtcact cctggcagtg tccccactgg gcctaccatc aacaccacca
     1381 tccaggatgt caaccgctac ctcctcaaga gtggaggtaa actgtcagac atcaaatcga
     1441 cgtggtcctc tggccctacc tcccacacgc aagcctctct gtctcatgaa ctatggaagg
     1501 tgcccagaaa cagtactgca cccacgaggc cacctccagg gttaaccaat cccaagccct
     1561 cctccacctg gggtgccagc cccctcggct ggaccagctc ctactcctcg ggttctgcct
     1621 ggagcaccga cacctcagga agaaccagca gctggctcgt tcttcgaaac ctcactcccc
     1681 agattgatgg ttctacactg cggacattgt gtttgcaaca tgggcctctt atcacattcc
     1741 acctgaatct gactcaaggc aatgctgtgg tccggtacag ctccaaggag gaggctgcca
     1801 aggcccagaa gtctctgcac atgtgcgtcc tgggaaacac taccatcctg gccgagttcg
     1861 ctggtgaaga agaagtgaat cgcttcttag cccaaggcca ggcgctgcca cccacttcca
     1921 gctggcagtc cagcagcgcg tccagccagc cgcggctcag cgcagcgggc agctcccatg
     1981 gcctggtacg cagcgacgct ggccactgga acgccccgtg cctgggtggc aaggggagca
     2041 gtgagctgct gtggggcggg gtgccccagt actccagcag cctgtggggc ccgcccagcg
     2101 ccgactgagc ggccgcatcg tgactgactg acgatctgcc tcgcgcgttt cggtgatgac
     2161 ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat
     2221 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca
     2281 gccatgaccc agtcacgtag cgatagcgga gtgtataatt cttgaagacg aaagggcctc
     2341 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     2401 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     2461 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     2521 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     2581 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     2641 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     2701 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     2761 ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     2821 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     2881 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     2941 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     3001 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     3061 acgatgcctg cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     3121 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     3181 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     3241 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     3301 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     3361 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     3421 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     3481 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     3541 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     3601 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     3661 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg
     3721 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     3781 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     3841 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     3901 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     3961 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     4021 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     4081 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     4141 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     4201 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     4261 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     4321 gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     4381 ataaattccg acaccatcga atggcgcaaa acctttcgcg gtatggcatg atagcgcccg
     4441 gaagagagtc aattcagggt ggtgaatgtg aaaccagtaa cgttatacga tgtcgcagag
     4501 tatgccggtg tctcttatca gaccgtttcc cgcgtggtga accaggccag ccacgtttct
     4561 gcgaaaacgc gggaaaaagt ggaagcggcg atggcggagc tgaattacat tcccaaccgc
     4621 gtggcacaac aactggcggg caaacagtcg ttgctgattg gcgttgccac ctccagtctg
     4681 gccctgcacg cgccgtcgca aattgtcgcg gcgattaaat ctcgcgccga tcaactgggt
     4741 gccagcgtgg tggtgtcgat ggtagaacga agcggcgtcg aagcctgtaa agcggcggtg
     4801 cacaatcttc tcgcgcaacg cgtcagtggg ctgatcatta actatccgct ggatgaccag
     4861 gatgccattg ctgtggaagc tgcctgcact aatgttccgg cgttatttct tgatgtctct
     4921 gaccagacac ccatcaacag tattattttc tcccatgaag acggtacgcg actgggcgtg
     4981 gagcatctgg tcgcattggg tcaccagcaa atcgcgctgt tagcgggccc attaagttct
     5041 gtctcggcgc gtctgcgtct ggctggctgg cataaatatc tcactcgcaa tcaaattcag
     5101 ccgatagcgg aacgggaagg cgactggagt gccatgtccg gttttcaaca aaccatgcaa
     5161 atgctgaatg agggcatcgt tcccactgcg atgctggttg ccaacgatca gatggcgctg
     5221 ggcgcaatgc gcgccattac cgagtccggg ctgcgcgttg gtgcggatat ctcggtagtg
     5281 ggatacgacg ataccgaaga cagctcatgt tatatcccgc cgttaaccac catcaaacag
     5341 gattttcgcc tgctggggca aaccagcgtg gaccgcttgc tgcaactctc tcagggccag
     5401 gcggtgaagg gcaatcagct gttgcccgtc tcactggtga aaagaaaaac cacgctagcg
     5461 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
     5521 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtaa gttagctcac
     5581 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt
     5641 gagcggataa caatttcaca caggaaacag ctatgaccat gattacggat tcactggccg
     5701 tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag
     5761 cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
     5821 aacagttgcg cagcctgaat ggcgaatggc gctttgcctg gtttccggca ccagaagcgg
     5881 tgccggaaag ctggctggag tgcgatcttc ctgaggccga tactgtcgtc gtcccctcaa
     5941 actggcagat gcacggttac gatgcgccca tctacaccaa cgtgacctat cccattacgg
     6001 tcaatccgcc gtttgttccc acggagaatc cgacgggttg ttactcgctc acatttaatg
     6061 ttgatgaaag ctggctacag gaaggccaga cgcgaattat ttttgatggc gttggaatt
//
