LOCUS       sequence-309115-        6606 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses XI in E.coli cell.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6606)
  AUTHORS   Han BG, Bong SM, Cho JW, Kim MD, Kim SJ, Lee BI
  TITLE     Crystal structure of a class 2 D-xylose isomerase from the human 
            intestinal tract microbe Bacteroides thetaiotaomicron
  JOURNAL   Korean Soc Struct Biology 2015; 3(1): 41-47
REFERENCE   2  (bases 1 to 6606)
  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: "Korean Soc 
            Struct Biology"; date: "2015"; volume: " 3(1)"; pages: " 41-4"
COMMENT     Sequence Label: pET28b-XI
FEATURES             Location/Qualifiers
     source          1..6606
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     48..67
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        48..66
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    67..91
                     /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             106..128
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             147..164
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             174..191
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1518..1535
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1588..1606)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1602..1649
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1686..2141
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1773..1792)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1983..2004
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2233..3048)
                     /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(2233..3048)
                     /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     2956..2975
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3040..3059
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3170..3758
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3659..3678
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3912..3929
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3944..4083
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4009..4028)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4169..4191
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4185..4376)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5185..6267)
                     /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     6229..6248
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6268..6345)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6551..6570
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 ccacgatgcg tccggcgtag aggatcgaga tctcgatccc gcgaaattaa tacgactcac
       61 tataggggaa ttgtgagcgg ataacaattc ccctctagaa ataattttgt ttaactttaa
      121 gaaggagata taccatgggc agcagccatc atcatcatca tcacagcagc ggcctggtgc
      181 cgcgcggcag ccatatggca acaaaagaat tttttccggg aattgaaaag attaaatttg
      241 aaggtaaaga tagtaagaac ccgatggcat tccgttatta cgatgcagag aaggtgatta
      301 atggtaaaaa gatgaaggat tggctgagat tcgctatggc atggtggcac acattgtgcg
      361 ctgaaggtgg tgatcagttc ggtggcggaa caaagcaatt cccatggaat ggtaatgcag
      421 atgctataca ggcagcaaaa gataagatgg atgcaggatt tgaattcatg cagaagatgg
      481 gtatcgaata ctattgcttc catgacgtag acttggtttc ggaaggtgcc agtgtagaag
      541 aatacgaagc taacctgaaa gaaatcgtag cttatgcaaa acagaaacag gcagaaaccg
      601 gtatcaaact actgtggggt actgctaatg tattcggtca cgcccgctat atgaacggtg
      661 cagctaccaa tcctgacttc gatgtagtag ctcgtgctgc tgttcagatc aaaaatgcga
      721 ttgatgcaac gattgaactt ggcggagaga attatgtgtt ttggggtggt cgtgaaggct
      781 atatgtctct tctgaacaca gatcagaaac gtgaaaaaga acaccttgca cagatgttga
      841 cgattgctcg tgactatgcc cgtgcccgtg gtttcaaagg tactttcctg atcgaaccga
      901 aaccgatgga accgactaaa catcaatatg acgtagatac ggaaactgta atcggcttcc
      961 tgaaagctca tggtctggat aaggatttca aagtaaatat cgaggtgaat cacgcaactt
     1021 tggcaggtca cactttcgag catgaattgg ctgtagctgt agacaatggt atgttgggct
     1081 caattgacgc caatcgtggt gactatcaga atggctggga tacagaccaa ttcccgatcg
     1141 acaattatga actgactcag gctatgatgc agattatccg taatggtggt ctcggtaccg
     1201 gtggtacgaa ctttgatgct aaaacccgtc gtaattctac tgatctggaa gatatcttta
     1261 ttgctcacat cgcaggtatg gacgctatgg cccgtgcact cgaaagtgca gcggctctgc
     1321 tcgacgaatc tccctataag aagatgctgg ctgaccgtta tgcttcattt gatgggggca
     1381 aaggtaaaga atttgaagac ggcaagctga ctctggagga tgtggttgct tatgcaaaaa
     1441 caaaaggcga accgaaacag actagcggca agcaagaact ttatgaggca attctgaata
     1501 tgtattgcta actcgagcac caccaccacc accactgaga tccggctgct aacaaagccc
     1561 gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa ccccttgggg
     1621 cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc ggattggcga
     1681 atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     1741 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     1801 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     1861 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     1921 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     1981 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     2041 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     2101 atttaacgcg aattttaaca aaatattaac gcttacaatt taggtggcac ttttcgggga
     2161 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     2221 atgaattaat tcttagaaaa actcatcgag catcaaatga aactgcaatt tattcatatc
     2281 aggattatca ataccatatt tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc
     2341 gaggcagttc cataggatgg caagatcctg gtatcggtct gcgattccga ctcgtccaac
     2401 atcaatacaa cctattaatt tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc
     2461 atgagtgacg actgaatccg gtgagaatgg caaaagttta tgcatttctt tccagacttg
     2521 ttcaacaggc cagccattac gctcgtcatc aaaatcactc gcatcaacca aaccgttatt
     2581 cattcgtgat tgcgcctgag cgagacgaaa tacgcgatcg ctgttaaaag gacaattaca
     2641 aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa tattttcacc
     2701 tgaatcagga tattcttcta atacctggaa tgctgttttc ccggggatcg cagtggtgag
     2761 taaccatgca tcatcaggag tacggataaa atgcttgatg gtcggaagag gcataaattc
     2821 cgtcagccag tttagtctga ccatctcatc tgtaacatca ttggcaacgc tacctttgcc
     2881 atgtttcaga aacaactctg gcgcatcggg cttcccatac aatcgataga ttgtcgcacc
     2941 tgattgcccg acattatcgc gagcccattt atacccatat aaatcagcat ccatgttgga
     3001 atttaatcgc ggcctagagc aagacgtttc ccgttgaata tggctcataa caccccttgt
     3061 attactgttt atgtaagcag acagttttat tgttcatgac caaaatccct taacgtgagt
     3121 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     3181 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     3241 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     3301 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     3361 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     3421 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     3481 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     3541 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     3601 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     3661 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     3721 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     3781 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     3841 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     3901 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc
     3961 tccttacgca tctgtgcggt atttcacacc gcaatggtgc actctcagta caatctgctc
     4021 tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg ggtcatggct
     4081 gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     4141 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     4201 tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat
     4261 tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag aagcgttaat
     4321 gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt ggtcactgat
     4381 gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatgaa acgagagagg
     4441 atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg ttgtgagggt
     4501 aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg tcaatgccag
     4561 cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc tgcgatgcag
     4621 atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta cgaaacacgg
     4681 aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt
     4741 cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc ccgccagcct
     4801 agccgggtcc tcaacgacag gagcacgatc atgcgcaccc gtggggccgc catgccggcg
     4861 ataatggcct gcttctcgcc gaaacgtttg gtggcgggac cagtgacgaa ggcttgagcg
     4921 agggcgtgca agattccgaa taccgcaagc gacaggccga tcatcgtcgc gctccagcga
     4981 aagcggtcct cgccgaaaat gacccagagc gctgccggca cctgtcctac gagttgcatg
     5041 ataaagaaga cagtcataag tgcggcgacg atagtcatgc cccgcgccca ccggaaggag
     5101 ctgactgggt tgaaggctct caagggcatc ggtcgagatc ccggtgccta atgagtgagc
     5161 taacttacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     5221 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgccag
     5281 ggtggttttt cttttcacca gtgagacggg caacagctga ttgcccttca ccgcctggcc
     5341 ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt
     5401 gatggtggtt aacggcggga tataacatga gctgtcttcg gtatcgtcgt atcccactac
     5461 cgagatatcc gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc
     5521 catctgatcg ttggcaacca gcatcgcagt gggaacgatg ccctcattca gcatttgcat
     5581 ggtttgttga aaaccggaca tggcactcca gtcgccttcc cgttccgcta tcggctgaat
     5641 ttgattgcga gtgagatatt tatgccagcc agccagacgc agacgcgccg agacagaact
     5701 taatgggccc gctaacagcg cgatttgctg gtgacccaat gcgaccagat gctccacgcc
     5761 cagtcgcgta ccgtcttcat gggagaaaat aatactgttg atgggtgtct ggtcagagac
     5821 atcaagaaat aacgccggaa cattagtgca ggcagcttcc acagcaatgg catcctggtc
     5881 atccagcgga tagttaatga tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc
     5941 cgctttacag gcttcgacgc cgcttcgttc taccatcgac accaccacgc tggcacccag
     6001 ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca gggccagact
     6061 ggaggtggca acgccaatca gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt
     6121 gggaatgtaa ttcagctccg ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac
     6181 gtggctggcc tggttcacca cgcgggaaac ggtctgataa gagacaccgg catactctgc
     6241 gacatcgtat aacgttactg gtttcacatt caccaccctg aattgactct cttccgggcg
     6301 ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg gtgtccggga tctcgacgct
     6361 ctcccttatg cgactcctgc attaggaagc agcccagtag taggttgagg ccgttgagca
     6421 ccgccgccgc aaggaatggt gcatgcaagg agatggcgcc caacagtccc ccggccacgg
     6481 ggcctgccac catacccacg ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat
     6541 cttccccatc ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg gcgccggtga
     6601 tgccgg
//
