LOCUS       sequence-197630-        5688 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of N-terminal region of human HRASLS3 in E.coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5688)
  AUTHORS   Han BG, Cho JW, Cho YD, Kim SY, Yoon HJ, Song HK, Cheong HK, Jeon 
            YH, Lee DK, Lee S, Lee BI
  TITLE     Expression, purification and biochemical characterization of the 
            N-terminal regions of human TIG3 and HRASLS3 proteins.
  JOURNAL   Protein Expr Purif. 2010 May;71(1):103-7. doi: 
            10.1016/j.pep.2010.01.018. Epub 2010 Jan 25.
  PUBMED    20100577
REFERENCE   2  (bases 1 to 5688)
  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"; doi: 
            "10.1016/j.pep.2010.01.018"; journalName: "Protein Expr Purif"; 
            date: "2010-05"; volume: "71"; issue: "1"; pages: "103-7"
COMMENT     Sequence Label: pET28b-HRASLS3-1-132
FEATURES             Location/Qualifiers
     source          1..5688
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     73..92
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     176..195
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        176..194
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    195..219
                     /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)."
     RBS             234..256
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             275..292
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             302..319
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             728..745
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(798..816)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      812..859
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      896..1351
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(983..1002)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1193..1214
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1443..2258)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(1443..2258)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2166..2185
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2250..2269
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2380..2968
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2869..2888
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3122..3139
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3154..3293
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3219..3238)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3379..3401
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3395..3586)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4395..5477)
                     /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     5439..5458
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5478..5555)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 ggggcctgcc accataccca cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg
       61 atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg tggcgccggt
      121 gatgccggcc acgatgcgtc cggcgtagag gatcgagatc tcgatcccgc gaaattaata
      181 cgactcacta taggggaatt gtgagcggat aacaattccc ctctagaaat aattttgttt
      241 aactttaaga aggagatata ccatgggcag cagccatcat catcatcatc acagcagcgg
      301 cctggtgccg cgcggcagcc atatgcgtgc gcccattcca gagcctaagc ctggagacct
      361 gattgagatt tttcgccctt tctacagaca ctgggccatc tatgttggcg atggatatgt
      421 ggttcatctg gcccctccaa gtgaggtcgc aggagctggt gcagccagtg tcatgtccgc
      481 cctgactgac aaggccatcg tgaagaagga attgctgtat gatgtggccg ggagtgacaa
      541 gtaccaggtc aacaacaaac atgatgacaa gtactcgccg ctgccctgca gcaaaatcat
      601 ccagcgggcg gaggagctgg tggggcagga ggtgctctac aagctgacca gtgagaactg
      661 cgagcacttt gtgaatgagc tgcgctatgg agtcgcccgc agtgaccagg tcagagatta
      721 actcgagcac caccaccacc accactgaga tccggctgct aacaaagccc gaaaggaagc
      781 tgagttggct gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg
      841 ggtcttgagg ggttttttgc tgaaaggagg aactatatcc ggattggcga atgggacgcg
      901 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
      961 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     1021 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     1081 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     1141 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     1201 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     1261 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     1321 aattttaaca aaatattaac gcttacaatt taggtggcac ttttcgggga aatgtgcgcg
     1381 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgaattaat
     1441 tcttagaaaa actcatcgag catcaaatga aactgcaatt tattcatatc aggattatca
     1501 ataccatatt tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc gaggcagttc
     1561 cataggatgg caagatcctg gtatcggtct gcgattccga ctcgtccaac atcaatacaa
     1621 cctattaatt tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc atgagtgacg
     1681 actgaatccg gtgagaatgg caaaagttta tgcatttctt tccagacttg ttcaacaggc
     1741 cagccattac gctcgtcatc aaaatcactc gcatcaacca aaccgttatt cattcgtgat
     1801 tgcgcctgag cgagacgaaa tacgcgatcg ctgttaaaag gacaattaca aacaggaatc
     1861 gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa tattttcacc tgaatcagga
     1921 tattcttcta atacctggaa tgctgttttc ccggggatcg cagtggtgag taaccatgca
     1981 tcatcaggag tacggataaa atgcttgatg gtcggaagag gcataaattc cgtcagccag
     2041 tttagtctga ccatctcatc tgtaacatca ttggcaacgc tacctttgcc atgtttcaga
     2101 aacaactctg gcgcatcggg cttcccatac aatcgataga ttgtcgcacc tgattgcccg
     2161 acattatcgc gagcccattt atacccatat aaatcagcat ccatgttgga atttaatcgc
     2221 ggcctagagc aagacgtttc ccgttgaata tggctcataa caccccttgt attactgttt
     2281 atgtaagcag acagttttat tgttcatgac caaaatccct taacgtgagt tttcgttcca
     2341 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     2401 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     2461 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     2521 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     2581 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     2641 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     2701 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     2761 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     2821 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     2881 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     2941 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     3001 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     3061 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     3121 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca
     3181 tctgtgcggt atttcacacc gcaatggtgc actctcagta caatctgctc tgatgccgca
     3241 tagttaagcc agtatacact ccgctatcgc tacgtgactg ggtcatggct gcgccccgac
     3301 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     3361 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     3421 aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat tcacagatgt
     3481 ctgcctgttc atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc
     3541 tgataaagcg ggccatgtta agggcggttt tttcctgttt ggtcactgat gcctccgtgt
     3601 aagggggatt tctgttcatg ggggtaatga taccgatgaa acgagagagg atgctcacga
     3661 tacgggttac tgatgatgaa catgcccggt tactggaacg ttgtgagggt aaacaactgg
     3721 cggtatggat gcggcgggac cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta
     3781 atacagatgt aggtgttcca cagggtagcc agcagcatcc tgcgatgcag atccggaaca
     3841 taatggtgca gggcgctgac ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga
     3901 ccattcatgt tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct
     3961 cgcgtatcgg tgattcattc tgctaaccag taaggcaacc ccgccagcct agccgggtcc
     4021 tcaacgacag gagcacgatc atgcgcaccc gtggggccgc catgccggcg ataatggcct
     4081 gcttctcgcc gaaacgtttg gtggcgggac cagtgacgaa ggcttgagcg agggcgtgca
     4141 agattccgaa taccgcaagc gacaggccga tcatcgtcgc gctccagcga aagcggtcct
     4201 cgccgaaaat gacccagagc gctgccggca cctgtcctac gagttgcatg ataaagaaga
     4261 cagtcataag tgcggcgacg atagtcatgc cccgcgccca ccggaaggag ctgactgggt
     4321 tgaaggctct caagggcatc ggtcgagatc ccggtgccta atgagtgagc taacttacat
     4381 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     4441 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgccag ggtggttttt
     4501 cttttcacca gtgagacggg caacagctga ttgcccttca ccgcctggcc ctgagagagt
     4561 tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt
     4621 aacggcggga tataacatga gctgtcttcg gtatcgtcgt atcccactac cgagatatcc
     4681 gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg
     4741 ttggcaacca gcatcgcagt gggaacgatg ccctcattca gcatttgcat ggtttgttga
     4801 aaaccggaca tggcactcca gtcgccttcc cgttccgcta tcggctgaat ttgattgcga
     4861 gtgagatatt tatgccagcc agccagacgc agacgcgccg agacagaact taatgggccc
     4921 gctaacagcg cgatttgctg gtgacccaat gcgaccagat gctccacgcc cagtcgcgta
     4981 ccgtcttcat gggagaaaat aatactgttg atgggtgtct ggtcagagac atcaagaaat
     5041 aacgccggaa cattagtgca ggcagcttcc acagcaatgg catcctggtc atccagcgga
     5101 tagttaatga tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag
     5161 gcttcgacgc cgcttcgttc taccatcgac accaccacgc tggcacccag ttgatcggcg
     5221 cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca gggccagact ggaggtggca
     5281 acgccaatca gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa
     5341 ttcagctccg ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac gtggctggcc
     5401 tggttcacca cgcgggaaac ggtctgataa gagacaccgg catactctgc gacatcgtat
     5461 aacgttactg gtttcacatt caccaccctg aattgactct cttccgggcg ctatcatgcc
     5521 ataccgcgaa aggttttgcg ccattcgatg gtgtccggga tctcgacgct ctcccttatg
     5581 cgactcctgc attaggaagc agcccagtag taggttgagg ccgttgagca ccgccgccgc
     5641 aaggaatggt gcatgcaagg agatggcgcc caacagtccc ccggccac
//
