LOCUS       sequence-164317-        5860 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  csd2, construct: 121-308, pET28b(+) NC-tag, mutation: S166A.
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,S166A/pET28b(+)
FEATURES             Location/Qualifiers
     source          1..5860
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      32..620
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     521..540
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     774..791
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    806..948
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(874..893)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1034..1056
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1050..1241)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(2050..3132)
                     /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     3094..3113
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3133..3210)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     3416..3435
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     3519..3538
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3519..3537
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3538..3562
                     /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             3577..3599
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3618..3635
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3645..3662
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             4239..4256
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4309..4327)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4323..4370
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      4407..4862
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4494..4513)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4704..4725
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4955..5770)
                     /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(4955..5770)
                     /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     5678..5697
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5762..5781
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
       61 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
      121 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
      181 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
      241 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
      301 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
      361 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
      421 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
      481 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
      541 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
      601 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
      661 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
      721 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
      781 cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg
      841 gtatttcaca ccgcatatat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
      901 aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc ccgacacccg
      961 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     1021 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     1081 gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca gatgtctgcc
     1141 tgttcatccg cgtccagctc gttgagtttc tccagaagcg ttaatgtctg gcttctgata
     1201 aagcgggcca tgttaagggc ggttttttcc tgtttggtca ctgatgcctc cgtgtaaggg
     1261 ggatttctgt tcatgggggt aatgataccg atgaaacgag agaggatgct cacgatacgg
     1321 gttactgatg atgaacatgc ccggttactg gaacgttgtg agggtaaaca actggcggta
     1381 tggatgcggc gggaccagag aaaaatcact cagggtcaat gccagcgctt cgttaataca
     1441 gatgtaggtg ttccacaggg tagccagcag catcctgcga tgcagatccg gaacataatg
     1501 gtgcagggcg ctgacttccg cgtttccaga ctttacgaaa cacggaaacc gaagaccatt
     1561 catgttgttg ctcaggtcgc agacgttttg cagcagcagt cgcttcacgt tcgctcgcgt
     1621 atcggtgatt cattctgcta accagtaagg caaccccgcc agcctagccg ggtcctcaac
     1681 gacaggagca cgatcatgcg cacccgtggg gccgccatgc cggcgataat ggcctgcttc
     1741 tcgccgaaac gtttggtggc gggaccagtg acgaaggctt gagcgagggc gtgcaagatt
     1801 ccgaataccg caagcgacag gccgatcatc gtcgcgctcc agcgaaagcg gtcctcgccg
     1861 aaaatgaccc agagcgctgc cggcacctgt cctacgagtt gcatgataaa gaagacagtc
     1921 ataagtgcgg cgacgatagt catgccccgc gcccaccgga aggagctgac tgggttgaag
     1981 gctctcaagg gcatcggtcg agatcccggt gcctaatgag tgagctaact tacattaatt
     2041 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     2101 atcggccaac gcgcggggag aggcggtttg cgtattgggc gccagggtgg tttttctttt
     2161 caccagtgag acgggcaaca gctgattgcc cttcaccgcc tggccctgag agagttgcag
     2221 caagcggtcc acgctggttt gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg
     2281 cgggatataa catgagctgt cttcggtatc gtcgtatccc actaccgaga tatccgcacc
     2341 aacgcgcagc ccggactcgg taatggcgcg cattgcgccc agcgccatct gatcgttggc
     2401 aaccagcatc gcagtgggaa cgatgccctc attcagcatt tgcatggttt gttgaaaacc
     2461 ggacatggca ctccagtcgc cttcccgttc cgctatcggc tgaatttgat tgcgagtgag
     2521 atatttatgc cagccagcca gacgcagacg cgccgagaca gaacttaatg ggcccgctaa
     2581 cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc
     2641 ttcatgggag aaaataatac tgttgatggg tgtctggtca gagacatcaa gaaataacgc
     2701 cggaacatta gtgcaggcag cttccacagc aatggcatcc tggtcatcca gcggatagtt
     2761 aatgatcagc ccactgacgc gttgcgcgag aagattgtgc accgccgctt tacaggcttc
     2821 gacgccgctt cgttctacca tcgacaccac cacgctggca cccagttgat cggcgcgaga
     2881 tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc agactggagg tggcaacgcc
     2941 aatcagcaac gactgtttgc ccgccagttg ttgtgccacg cggttgggaa tgtaattcag
     3001 ctccgccatc gccgcttcca ctttttcccg cgttttcgca gaaacgtggc tggcctggtt
     3061 caccacgcgg gaaacggtct gataagagac accggcatac tctgcgacat cgtataacgt
     3121 tactggtttc acattcacca ccctgaattg actctcttcc gggcgctatc atgccatacc
     3181 gcgaaaggtt ttgcgccatt cgatggtgtc cgggatctcg acgctctccc ttatgcgact
     3241 cctgcattag gaagcagccc agtagtaggt tgaggccgtt gagcaccgcc gccgcaagga
     3301 atggtgcatg caaggagatg gcgcccaaca gtcccccggc cacggggcct gccaccatac
     3361 ccacgccgaa acaagcgctc atgagcccga agtggcgagc ccgatcttcc ccatcggtga
     3421 tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc
     3481 gtccggcgta gaggatcgag atctcgatcc cgcgaaatta atacgactca ctatagggga
     3541 attgtgagcg gataacaatt cccctctaga aataattttg tttaacttta agaaggagat
     3601 ataccatggg cagcagccat catcatcatc atcacagcag cggcctggtg ccgcgcggca
     3661 gccatatgtt agacaatttg aatctggctc aaaaacattt agcgctcatg ctcattccta
     3721 acggcatgcc cataaaaact tatagcgcta tcaaacccac caaagaaagg aaccacccca
     3781 ttaaaaaaat taagggcgtt gaagccggga ttgattttat cgcgccatta aacacgcctg
     3841 tttatgcgag cgctgatggg attgtggatt ttgtgaagac caattctaat gtggggtatg
     3901 ggaacttggt gcgcattgaa catgcgtttg gttttagctc catttacacg cacttagatc
     3961 atgtcaatgt gcagcccaaa agcttcatcc aaaaagggca gttgattggc tatagcggga
     4021 agagcggtaa tagcggcggc gaaaaattgc attatgaagt gcgctttttg ggtaaaattt
     4081 tagacgcgca aaaatttctg gcatgggatt tggatcattt tcaaagcgct ttagaagaaa
     4141 ataaatttat tgaatggaag aatttgtttt gggttttaga agacattgtc cagctccaag
     4201 agcatgtgga taaagacgca ctcataagcc agctcgagca ccaccaccac caccactgag
     4261 atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat
     4321 aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag
     4381 gaactatatc cggattggcg aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg
     4441 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     4501 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     4561 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     4621 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     4681 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     4741 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     4801 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat
     4861 ttcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     4921 acattcaaat atgtatccgc tcatgaatta attcttagaa aaactcatcg agcatcaaat
     4981 gaaactgcaa tttattcata tcaggattat caataccata tttttgaaaa agccgtttct
     5041 gtaatgaagg agaaaactca ccgaggcagt tccataggat ggcaagatcc tggtatcggt
     5101 ctgcgattcc gactcgtcca acatcaatac aacctattaa tttcccctcg tcaaaaataa
     5161 ggttatcaag tgagaaatca ccatgagtga cgactgaatc cggtgagaat ggcaaaagtt
     5221 tatgcatttc tttccagact tgttcaacag gccagccatt acgctcgtca tcaaaatcac
     5281 tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg agcgagacga aatacgcgat
     5341 cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg aacactgcca
     5401 gcgcatcaac aatattttca cctgaatcag gatattcttc taatacctgg aatgctgttt
     5461 tcccggggat cgcagtggtg agtaaccatg catcatcagg agtacggata aaatgcttga
     5521 tggtcggaag aggcataaat tccgtcagcc agtttagtct gaccatctca tctgtaacat
     5581 cattggcaac gctacctttg ccatgtttca gaaacaactc tggcgcatcg ggcttcccat
     5641 acaatcgata gattgtcgca cctgattgcc cgacattatc gcgagcccat ttatacccat
     5701 ataaatcagc atccatgttg gaatttaatc gcggcctaga gcaagacgtt tcccgttgaa
     5761 tatggctcat aacacccctt gtattactgt ttatgtaagc agacagtttt attgttcatg
     5821 accaaaatcc cttaacgtga gttttcgttc cactgagcgt
//
