LOCUS       sequence-164300-        5860 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  csd2, construct: 121-308, pET28b(+) NC-tag, mutation: Y197F.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5860)
  AUTHORS   An DR, Im HN, Jang JY, Kim HS, Kim J, Yoon HJ, Hesek D, Lee M, 
            Mobashery S, Kim SJ, Suh SW
  TITLE     Structural Basis of the Heterodimer Formation between Cell 
            Shape-Determining Proteins Csd1 and Csd2 from Helicobacter pylori.
  JOURNAL   PLoS One. 2016 Oct 6;11(10):e0164243. doi: 
            10.1371/journal.pone.0164243. eCollection 2016.
  PUBMED    27711177
REFERENCE   2  (bases 1 to 5860)
  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: "PLoS One.";
            date: "2016-10-6"; volume: "11(10):e0164243. doi"; pages: " 
            10.1371/journal.pone.0164243. eCollection 2016"
COMMENT     Sequence Label: csd2_121-308,Y197F/pET28b(+)
FEATURES             Location/Qualifiers
     source          1..5860
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      18..606
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     507..526
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     760..777
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    792..934
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(860..879)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1020..1042
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1036..1227)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(2036..3118)
                     /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     3080..3099
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3119..3196)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     3402..3421
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     3505..3524
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3505..3523
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3524..3548
                     /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             3563..3585
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3604..3621
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3631..3648
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             4225..4242
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4295..4313)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4309..4356
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      4393..4848
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4480..4499)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4690..4711
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4941..5756)
                     /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(4941..5756)
                     /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     5664..5683
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5748..5767
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
       61 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
      121 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg
      181 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
      241 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
      301 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
      361 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
      421 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
      481 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
      541 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
      601 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
      661 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
      721 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
      781 gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
      841 atatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatacac
      901 tccgctatcg ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga
      961 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     1021 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg
     1081 gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc
     1141 cagctcgttg agtttctcca gaagcgttaa tgtctggctt ctgataaagc gggccatgtt
     1201 aagggcggtt ttttcctgtt tggtcactga tgcctccgtg taagggggat ttctgttcat
     1261 gggggtaatg ataccgatga aacgagagag gatgctcacg atacgggtta ctgatgatga
     1321 acatgcccgg ttactggaac gttgtgaggg taaacaactg gcggtatgga tgcggcggga
     1381 ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc
     1441 acagggtagc cagcagcatc ctgcgatgca gatccggaac ataatggtgc agggcgctga
     1501 cttccgcgtt tccagacttt acgaaacacg gaaaccgaag accattcatg ttgttgctca
     1561 ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt
     1621 ctgctaacca gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca ggagcacgat
     1681 catgcgcacc cgtggggccg ccatgccggc gataatggcc tgcttctcgc cgaaacgttt
     1741 ggtggcggga ccagtgacga aggcttgagc gagggcgtgc aagattccga ataccgcaag
     1801 cgacaggccg atcatcgtcg cgctccagcg aaagcggtcc tcgccgaaaa tgacccagag
     1861 cgctgccggc acctgtccta cgagttgcat gataaagaag acagtcataa gtgcggcgac
     1921 gatagtcatg ccccgcgccc accggaagga gctgactggg ttgaaggctc tcaagggcat
     1981 cggtcgagat cccggtgcct aatgagtgag ctaacttaca ttaattgcgt tgcgctcact
     2041 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     2101 ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg
     2161 gcaacagctg attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc
     2221 tggtttgccc cagcaggcga aaatcctgtt tgatggtggt taacggcggg atataacatg
     2281 agctgtcttc ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg cgcagcccgg
     2341 actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc agcatcgcag
     2401 tgggaacgat gccctcattc agcatttgca tggtttgttg aaaaccggac atggcactcc
     2461 agtcgccttc ccgttccgct atcggctgaa tttgattgcg agtgagatat ttatgccagc
     2521 cagccagacg cagacgcgcc gagacagaac ttaatgggcc cgctaacagc gcgatttgct
     2581 ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa
     2641 taatactgtt gatgggtgtc tggtcagaga catcaagaaa taacgccgga acattagtgc
     2701 aggcagcttc cacagcaatg gcatcctggt catccagcgg atagttaatg atcagcccac
     2761 tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt
     2821 ctaccatcga caccaccacg ctggcaccca gttgatcggc gcgagattta atcgccgcga
     2881 caatttgcga cggcgcgtgc agggccagac tggaggtggc aacgccaatc agcaacgact
     2941 gtttgcccgc cagttgttgt gccacgcggt tgggaatgta attcagctcc gccatcgccg
     3001 cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa
     3061 cggtctgata agagacaccg gcatactctg cgacatcgta taacgttact ggtttcacat
     3121 tcaccaccct gaattgactc tcttccgggc gctatcatgc cataccgcga aaggttttgc
     3181 gccattcgat ggtgtccggg atctcgacgc tctcccttat gcgactcctg cattaggaag
     3241 cagcccagta gtaggttgag gccgttgagc accgccgccg caaggaatgg tgcatgcaag
     3301 gagatggcgc ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa
     3361 gcgctcatga gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag
     3421 gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg
     3481 atcgagatct cgatcccgcg aaattaatac gactcactat aggggaattg tgagcggata
     3541 acaattcccc tctagaaata attttgttta actttaagaa ggagatatac catgggcagc
     3601 agccatcatc atcatcatca cagcagcggc ctggtgccgc gcggcagcca tatgttagac
     3661 aatttgaatc tggctcaaaa acatttagcg ctcatgctca ttcctaacgg catgcccata
     3721 aaaacttata gcgctatcaa acccaccaaa gaaaggaacc accccattaa aaaaattaag
     3781 ggcgttgaat ccgggattga ttttatcgcg ccattaaaca cgcctgttta tgcgagcgct
     3841 gatgggattg tggattttgt gaagaccaat tctaatgtgg ggtttgggaa cttggtgcgc
     3901 attgaacatg cgtttggttt tagctccatt tacacgcact tagatcatgt caatgtgcag
     3961 cccaaaagct tcatccaaaa agggcagttg attggctata gcgggaagag cggtaatagc
     4021 ggcggcgaaa aattgcatta tgaagtgcgc tttttgggta aaattttaga cgcgcaaaaa
     4081 tttctggcat gggatttgga tcattttcaa agcgctttag aagaaaataa atttattgaa
     4141 tggaagaatt tgttttgggt tttagaagac attgtccagc tccaagagca tgtggataaa
     4201 gacgcactca taagccagct cgagcaccac caccaccacc actgagatcc ggctgctaac
     4261 aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc
     4321 cttggggcct ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatccgga
     4381 ttggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     4441 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     4501 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     4561 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     4621 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     4681 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     4741 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     4801 aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttca ggtggcactt
     4861 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     4921 atccgctcat gaattaattc ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta
     4981 ttcatatcag gattatcaat accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa
     5041 aactcaccga ggcagttcca taggatggca agatcctggt atcggtctgc gattccgact
     5101 cgtccaacat caatacaacc tattaatttc ccctcgtcaa aaataaggtt atcaagtgag
     5161 aaatcaccat gagtgacgac tgaatccggt gagaatggca aaagtttatg catttctttc
     5221 cagacttgtt caacaggcca gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa
     5281 ccgttattca ttcgtgattg cgcctgagcg agacgaaata cgcgatcgct gttaaaagga
     5341 caattacaaa caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata
     5401 ttttcacctg aatcaggata ttcttctaat acctggaatg ctgttttccc ggggatcgca
     5461 gtggtgagta accatgcatc atcaggagta cggataaaat gcttgatggt cggaagaggc
     5521 ataaattccg tcagccagtt tagtctgacc atctcatctg taacatcatt ggcaacgcta
     5581 cctttgccat gtttcagaaa caactctggc gcatcgggct tcccatacaa tcgatagatt
     5641 gtcgcacctg attgcccgac attatcgcga gcccatttat acccatataa atcagcatcc
     5701 atgttggaat ttaatcgcgg cctagagcaa gacgtttccc gttgaatatg gctcataaca
     5761 ccccttgtat tactgtttat gtaagcagac agttttattg ttcatgacca aaatccctta
     5821 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
//
