LOCUS       sequence-197633-        5784 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Full-length human TIG3 in E.coli expression vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5784)
  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 5784)
  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-hTIG3-1-164
FEATURES             Location/Qualifiers
     source          1..5784
                     /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             824..841
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(894..912)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      908..955
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      992..1447
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1079..1098)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1289..1310
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1539..2354)
                     /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(1539..2354)
                     /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     2262..2281
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2346..2365
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2476..3064
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2965..2984
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3218..3235
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3250..3389
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3315..3334)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3475..3497
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3491..3682)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4491..5573)
                     /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
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGACVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     5535..5554
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5574..5651)
                     /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 atatggcttc gccacaccaa gagcccaaac ctggagacct
      361 gattgagatt ttccgccttg gctatgagca ctgggccctg tatataggag atggctacgt
      421 gatccatctg gctcctccaa gtgagtaccc cggggctggc tcctccagtg tcttctcagt
      481 cctgagcaac agtgcagagg tgaaacggga gcgcctggaa gatgtggtgg gaggctgttg
      541 ctatcgggtc aacaacagct tggaccatga gtaccaacca cggcccgtgg aggtgatcat
      601 cagttctgcg aaggagatgg ttggtcagaa gatgaagtac agtattgtga gcaggaactg
      661 tgagcacttt gtcacccagc tgagatatgg caagtcccgc tgtaaacagg tggaaaaggc
      721 caaggttgaa gtcggtgtgg ccacggcgct tggaatcctg gttgttgctg gatgctcttt
      781 tgcgattagg agataccaaa aaaaagcaac agcctgactc gagcaccacc accaccacca
      841 ctgagatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga
      901 gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa
      961 aggaggaact atatccggat tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg
     1021 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     1081 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     1141 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     1201 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     1261 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     1321 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     1381 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt
     1441 acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     1501 aaatacattc aaatatgtat ccgctcatga attaattctt agaaaaactc atcgagcatc
     1561 aaatgaaact gcaatttatt catatcagga ttatcaatac catatttttg aaaaagccgt
     1621 ttctgtaatg aaggagaaaa ctcaccgagg cagttccata ggatggcaag atcctggtat
     1681 cggtctgcga ttccgactcg tccaacatca atacaaccta ttaatttccc ctcgtcaaaa
     1741 ataaggttat caagtgagaa atcaccatga gtgacgactg aatccggtga gaatggcaaa
     1801 agtttatgca tttctttcca gacttgttca acaggccagc cattacgctc gtcatcaaaa
     1861 tcactcgcat caaccaaacc gttattcatt cgtgattgcg cctgagcgag acgaaatacg
     1921 cgatcgctgt taaaaggaca attacaaaca ggaatcgaat gcaaccggcg caggaacact
     1981 gccagcgcat caacaatatt ttcacctgaa tcaggatatt cttctaatac ctggaatgct
     2041 gttttcccgg ggatcgcagt ggtgagtaac catgcatcat caggagtacg gataaaatgc
     2101 ttgatggtcg gaagaggcat aaattccgtc agccagttta gtctgaccat ctcatctgta
     2161 acatcattgg caacgctacc tttgccatgt ttcagaaaca actctggcgc atcgggcttc
     2221 ccatacaatc gatagattgt cgcacctgat tgcccgacat tatcgcgagc ccatttatac
     2281 ccatataaat cagcatccat gttggaattt aatcgcggcc tagagcaaga cgtttcccgt
     2341 tgaatatggc tcataacacc ccttgtatta ctgtttatgt aagcagacag ttttattgtt
     2401 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     2461 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     2521 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     2581 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta
     2641 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     2701 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     2761 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     2821 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     2881 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     2941 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     3001 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     3061 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     3121 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     3181 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     3241 ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcaa
     3301 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta tacactccgc
     3361 tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc gctgacgcgc
     3421 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     3481 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa
     3541 gctcatcagc gtggtcgtga agcgattcac agatgtctgc ctgttcatcc gcgtccagct
     3601 cgttgagttt ctccagaagc gttaatgtct ggcttctgat aaagcgggcc atgttaaggg
     3661 cggttttttc ctgtttggtc actgatgcct ccgtgtaagg gggatttctg ttcatggggg
     3721 taatgatacc gatgaaacga gagaggatgc tcacgatacg ggttactgat gatgaacatg
     3781 cccggttact ggaacgttgt gagggtaaac aactggcggt atggatgcgg cgggaccaga
     3841 gaaaaatcac tcagggtcaa tgccagcgct tcgttaatac agatgtaggt gttccacagg
     3901 gtagccagca gcatcctgcg atgcagatcc ggaacataat ggtgcagggc gctgacttcc
     3961 gcgtttccag actttacgaa acacggaaac cgaagaccat tcatgttgtt gctcaggtcg
     4021 cagacgtttt gcagcagcag tcgcttcacg ttcgctcgcg tatcggtgat tcattctgct
     4081 aaccagtaag gcaaccccgc cagcctagcc gggtcctcaa cgacaggagc acgatcatgc
     4141 gcacccgtgg ggccgccatg ccggcgataa tggcctgctt ctcgccgaaa cgtttggtgg
     4201 cgggaccagt gacgaaggct tgagcgaggg cgtgcaagat tccgaatacc gcaagcgaca
     4261 ggccgatcat cgtcgcgctc cagcgaaagc ggtcctcgcc gaaaatgacc cagagcgctg
     4321 ccggcacctg tcctacgagt tgcatgataa agaagacagt cataagtgcg gcgacgatag
     4381 tcatgccccg cgcccaccgg aaggagctga ctgggttgaa ggctctcaag ggcatcggtc
     4441 gagatcccgg tgcctaatga gtgagctaac ttacattaat tgcgttgcgc tcactgcccg
     4501 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     4561 gaggcggttt gcgtattggg cgccagggtg gtttttcttt tcaccagtga gacgggcaac
     4621 agctgattgc ccttcaccgc ctggccctga gagagttgca gcaagcggtc cacgctggtt
     4681 tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata acatgagctg
     4741 tcttcggtat cgtcgtatcc cactaccgag atatccgcac caacgcgcag cccggactcg
     4801 gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg caaccagcat cgcagtggga
     4861 acgatgccct cattcagcat ttgcatggtt tgttgaaaac cggacatggc actccagtcg
     4921 ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga gatatttatg ccagccagcc
     4981 agacgcagac gcgccgagac agaacttaat gggcccgcta acagcgcgat ttgctggtga
     5041 cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt cttcatggga gaaaataata
     5101 ctgttgatgg gtgtctggtc agagacatca agaaataacg ccggaacatt agtgcaggca
     5161 gcttccacag caatggcatc ctggtcatcc agcggatagt taatgatcag cccactgacg
     5221 cgttgcgcga gaagattgtg caccgccgct ttacaggctt cgacgccgct tcgttctacc
     5281 atcgacacca ccacgcaggc acccagttga tcggcgcgag atttaatcgc cgcgacaatt
     5341 tgcgacggcg cgtgcagggc cagactggag gtggcaacgc caatcagcaa cgactgtttg
     5401 cccgccagtt gttgtgccac gcggttggga atgtaattca gctccgccat cgccgcttcc
     5461 actttttccc gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc
     5521 tgataagaga caccggcata ctctgcgaca tcgtataacg ttactggttt cacattcacc
     5581 accctgaatt gactctcttc cgggcgctat catgccatac cgcgaaaggt tttgcgccat
     5641 tcgatggtgt ccgggatctc gacgctctcc cttatgcgac tcctgcatta ggaagcagcc
     5701 cagtagtagg ttgaggccgt tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat
     5761 ggcgcccaac agtcccccgg ccac
//
