LOCUS       sequence-164289-        5860 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  csd2, construct: 121-308, pET28b(+) NC-tag.
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/pET28b(+)
FEATURES             Location/Qualifiers
     source          1..5860
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      8..596
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     497..516
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     750..767
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    782..924
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(850..869)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1010..1032
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1026..1217)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(2026..3108)
                     /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     3070..3089
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3109..3186)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     3392..3411
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     3495..3514
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3495..3513
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3514..3538
                     /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             3553..3575
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3594..3611
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3621..3638
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             4215..4232
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4285..4303)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4299..4346
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      4383..4838
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4470..4489)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4680..4701
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4931..5746)
                     /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(4931..5746)
                     /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     5654..5673
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5738..5757
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
       61 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
      121 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
      181 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
      241 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
      301 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
      361 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
      421 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
      481 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
      541 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
      601 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
      661 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
      721 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
      781 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atatatggtg
      841 cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg
      901 ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga
      961 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     1021 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca
     1081 tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc cagctcgttg
     1141 agtttctcca gaagcgttaa tgtctggctt ctgataaagc gggccatgtt aagggcggtt
     1201 ttttcctgtt tggtcactga tgcctccgtg taagggggat ttctgttcat gggggtaatg
     1261 ataccgatga aacgagagag gatgctcacg atacgggtta ctgatgatga acatgcccgg
     1321 ttactggaac gttgtgaggg taaacaactg gcggtatgga tgcggcggga ccagagaaaa
     1381 atcactcagg gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc acagggtagc
     1441 cagcagcatc ctgcgatgca gatccggaac ataatggtgc agggcgctga cttccgcgtt
     1501 tccagacttt acgaaacacg gaaaccgaag accattcatg ttgttgctca ggtcgcagac
     1561 gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca
     1621 gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca ggagcacgat catgcgcacc
     1681 cgtggggccg ccatgccggc gataatggcc tgcttctcgc cgaaacgttt ggtggcggga
     1741 ccagtgacga aggcttgagc gagggcgtgc aagattccga ataccgcaag cgacaggccg
     1801 atcatcgtcg cgctccagcg aaagcggtcc tcgccgaaaa tgacccagag cgctgccggc
     1861 acctgtccta cgagttgcat gataaagaag acagtcataa gtgcggcgac gatagtcatg
     1921 ccccgcgccc accggaagga gctgactggg ttgaaggctc tcaagggcat cggtcgagat
     1981 cccggtgcct aatgagtgag ctaacttaca ttaattgcgt tgcgctcact gcccgctttc
     2041 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     2101 ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg gcaacagctg
     2161 attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc tggtttgccc
     2221 cagcaggcga aaatcctgtt tgatggtggt taacggcggg atataacatg agctgtcttc
     2281 ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg cgcagcccgg actcggtaat
     2341 ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc agcatcgcag tgggaacgat
     2401 gccctcattc agcatttgca tggtttgttg aaaaccggac atggcactcc agtcgccttc
     2461 ccgttccgct atcggctgaa tttgattgcg agtgagatat ttatgccagc cagccagacg
     2521 cagacgcgcc gagacagaac ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa
     2581 tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa taatactgtt
     2641 gatgggtgtc tggtcagaga catcaagaaa taacgccgga acattagtgc aggcagcttc
     2701 cacagcaatg gcatcctggt catccagcgg atagttaatg atcagcccac tgacgcgttg
     2761 cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga
     2821 caccaccacg ctggcaccca gttgatcggc gcgagattta atcgccgcga caatttgcga
     2881 cggcgcgtgc agggccagac tggaggtggc aacgccaatc agcaacgact gtttgcccgc
     2941 cagttgttgt gccacgcggt tgggaatgta attcagctcc gccatcgccg cttccacttt
     3001 ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa cggtctgata
     3061 agagacaccg gcatactctg cgacatcgta taacgttact ggtttcacat tcaccaccct
     3121 gaattgactc tcttccgggc gctatcatgc cataccgcga aaggttttgc gccattcgat
     3181 ggtgtccggg atctcgacgc tctcccttat gcgactcctg cattaggaag cagcccagta
     3241 gtaggttgag gccgttgagc accgccgccg caaggaatgg tgcatgcaag gagatggcgc
     3301 ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga
     3361 gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa
     3421 ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg atcgagatct
     3481 cgatcccgcg aaattaatac gactcactat aggggaattg tgagcggata acaattcccc
     3541 tctagaaata attttgttta actttaagaa ggagatatac catgggcagc agccatcatc
     3601 atcatcatca cagcagcggc ctggtgccgc gcggcagcca tatgttagac aatttgaatc
     3661 tggctcaaaa acatttagcg ctcatgctca ttcctaacgg catgcccata aaaacttata
     3721 gcgctatcaa acccaccaaa gaaaggaacc accccattaa aaaaattaag ggcgttgaat
     3781 ccgggattga ttttatcgcg ccattaaaca cgcctgttta tgcgagcgct gatgggattg
     3841 tggattttgt gaagaccaat tctaatgtgg ggtatgggaa cttggtgcgc attgaacatg
     3901 cgtttggttt tagctccatt tacacgcact tagatcatgt caatgtgcag cccaaaagct
     3961 tcatccaaaa agggcagttg attggctata gcgggaagag cggtaatagc ggcggcgaaa
     4021 aattgcatta tgaagtgcgc tttttgggta aaattttaga cgcgcaaaaa tttctggcat
     4081 gggatttgga tcattttcaa agcgctttag aagaaaataa atttattgaa tggaagaatt
     4141 tgttttgggt tttagaagac attgtccagc tccaagagca tgtggataaa gacgcactca
     4201 taagccagct cgagcaccac caccaccacc actgagatcc ggctgctaac aaagcccgaa
     4261 aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc cttggggcct
     4321 ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatccgga ttggcgaatg
     4381 ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     4441 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     4501 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     4561 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     4621 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     4681 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     4741 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     4801 taacgcgaat tttaacaaaa tattaacgtt tacaatttca ggtggcactt ttcggggaaa
     4861 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     4921 gaattaattc ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag
     4981 gattatcaat accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga
     5041 ggcagttcca taggatggca agatcctggt atcggtctgc gattccgact cgtccaacat
     5101 caatacaacc tattaatttc ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat
     5161 gagtgacgac tgaatccggt gagaatggca aaagtttatg catttctttc cagacttgtt
     5221 caacaggcca gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca
     5281 ttcgtgattg cgcctgagcg agacgaaata cgcgatcgct gttaaaagga caattacaaa
     5341 caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata ttttcacctg
     5401 aatcaggata ttcttctaat acctggaatg ctgttttccc ggggatcgca gtggtgagta
     5461 accatgcatc atcaggagta cggataaaat gcttgatggt cggaagaggc ataaattccg
     5521 tcagccagtt tagtctgacc atctcatctg taacatcatt ggcaacgcta cctttgccat
     5581 gtttcagaaa caactctggc gcatcgggct tcccatacaa tcgatagatt gtcgcacctg
     5641 attgcccgac attatcgcga gcccatttat acccatataa atcagcatcc atgttggaat
     5701 ttaatcgcgg cctagagcaa gacgtttccc gttgaatatg gctcataaca ccccttgtat
     5761 tactgtttat gtaagcagac agttttattg ttcatgacca aaatccctta acgtgagttt
     5821 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
//
