LOCUS       sequence-164294-        5863 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  csd2, construct: 121-308, pET28b(+) NC-tag, mutation: E165A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5863)
  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 5863)
  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,E165A/pET28b(+)
FEATURES             Location/Qualifiers
     source          1..5863
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      14..602
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     503..522
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     756..773
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    788..930
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(856..875)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1016..1038
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1032..1223)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(2032..3114)
                     /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     3076..3095
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3115..3192)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     3398..3417
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     3501..3520
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3501..3519
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3520..3544
                     /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             3559..3581
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3600..3617
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3627..3644
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             4221..4241
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     primer_bind     complement(4294..4312)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4308..4355
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      4392..4847
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4479..4498)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4689..4710
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4940..5755)
                     /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(4940..5755)
                     /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     5663..5682
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5747..5766
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
       61 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
      121 aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt
      181 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
      241 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
      301 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
      361 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
      421 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
      481 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
      541 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
      601 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
      661 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
      721 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
      781 aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatat
      841 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg
      901 ctatcgctac gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg
      961 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     1021 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa
     1081 agctcatcag cgtggtcgtg aagcgattca cagatgtctg cctgttcatc cgcgtccagc
     1141 tcgttgagtt tctccagaag cgttaatgtc tggcttctga taaagcgggc catgttaagg
     1201 gcggtttttt cctgtttggt cactgatgcc tccgtgtaag ggggatttct gttcatgggg
     1261 gtaatgatac cgatgaaacg agagaggatg ctcacgatac gggttactga tgatgaacat
     1321 gcccggttac tggaacgttg tgagggtaaa caactggcgg tatggatgcg gcgggaccag
     1381 agaaaaatca ctcagggtca atgccagcgc ttcgttaata cagatgtagg tgttccacag
     1441 ggtagccagc agcatcctgc gatgcagatc cggaacataa tggtgcaggg cgctgacttc
     1501 cgcgtttcca gactttacga aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc
     1561 gcagacgttt tgcagcagca gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc
     1621 taaccagtaa ggcaaccccg ccagcctagc cgggtcctca acgacaggag cacgatcatg
     1681 cgcacccgtg gggccgccat gccggcgata atggcctgct tctcgccgaa acgtttggtg
     1741 gcgggaccag tgacgaaggc ttgagcgagg gcgtgcaaga ttccgaatac cgcaagcgac
     1801 aggccgatca tcgtcgcgct ccagcgaaag cggtcctcgc cgaaaatgac ccagagcgct
     1861 gccggcacct gtcctacgag ttgcatgata aagaagacag tcataagtgc ggcgacgata
     1921 gtcatgcccc gcgcccaccg gaaggagctg actgggttga aggctctcaa gggcatcggt
     1981 cgagatcccg gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg ctcactgccc
     2041 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     2101 agaggcggtt tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg agacgggcaa
     2161 cagctgattg cccttcaccg cctggccctg agagagttgc agcaagcggt ccacgctggt
     2221 ttgccccagc aggcgaaaat cctgtttgat ggtggttaac ggcgggatat aacatgagct
     2281 gtcttcggta tcgtcgtatc ccactaccga gatatccgca ccaacgcgca gcccggactc
     2341 ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg
     2401 aacgatgccc tcattcagca tttgcatggt ttgttgaaaa ccggacatgg cactccagtc
     2461 gccttcccgt tccgctatcg gctgaatttg attgcgagtg agatatttat gccagccagc
     2521 cagacgcaga cgcgccgaga cagaacttaa tgggcccgct aacagcgcga tttgctggtg
     2581 acccaatgcg accagatgct ccacgcccag tcgcgtaccg tcttcatggg agaaaataat
     2641 actgttgatg ggtgtctggt cagagacatc aagaaataac gccggaacat tagtgcaggc
     2701 agcttccaca gcaatggcat cctggtcatc cagcggatag ttaatgatca gcccactgac
     2761 gcgttgcgcg agaagattgt gcaccgccgc tttacaggct tcgacgccgc ttcgttctac
     2821 catcgacacc accacgctgg cacccagttg atcggcgcga gatttaatcg ccgcgacaat
     2881 ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca acgactgttt
     2941 gcccgccagt tgttgtgcca cgcggttggg aatgtaattc agctccgcca tcgccgcttc
     3001 cactttttcc cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt
     3061 ctgataagag acaccggcat actctgcgac atcgtataac gttactggtt tcacattcac
     3121 caccctgaat tgactctctt ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca
     3181 ttcgatggtg tccgggatct cgacgctctc ccttatgcga ctcctgcatt aggaagcagc
     3241 ccagtagtag gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca tgcaaggaga
     3301 tggcgcccaa cagtcccccg gccacggggc ctgccaccat acccacgccg aaacaagcgc
     3361 tcatgagccc gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc
     3421 cagcaaccgc acctgtggcg ccggtgatgc cggccacgat gcgtccggcg tagaggatcg
     3481 agatctcgat cccgcgaaat taatacgact cactataggg gaattgtgag cggataacaa
     3541 ttcccctcta gaaataattt tgtttaactt taagaaggag atataccatg ggcagcagcc
     3601 atcatcatca tcatcacagc agcggcctgg tgccgcgcgg cagccatatg ttagacaatt
     3661 tgaatctggc tcaaaaacat ttagcgctca tgctcattcc taacggcatg cccataaaaa
     3721 cttatagcgc tatcaaaccc accaaagaaa ggaaccaccc cattaaaaaa attaagggcg
     3781 ttgcatccgg gattgatttt atcgcgccat taaacacgcc tgtttatgcg agcgctgatg
     3841 ggattgtgga ttttgtgaag accaattcta atgtggggta tgggaacttg gtgcgcattg
     3901 aacatgcgtt tggttttagc tccatttaca cgcacttaga tcatgtcaat gtgcagccca
     3961 aaagcttcat ccaaaaaggg cagttgattg gctatagcgg gaagagcggt aatagcggcg
     4021 gcgaaaaatt gcattatgaa gtgcgctttt tgggtaaaat tttagacgcg caaaaatttc
     4081 tggcatggga tttggatcat tttcaaagcg ctttagaaga aaataaattt attgaatgga
     4141 agaatttgtt ttgggtttta gaagacattg tccagctcca agagcatgtg gataaagacg
     4201 cactcataag ccagctcgag caccaccacc accaccacca ctgagatccg gctgctaaca
     4261 aagcccgaaa ggaagctgag ttggctgctg ccaccgctga gcaataacta gcataacccc
     4321 ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa aggaggaact atatccggat
     4381 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     4441 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     4501 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     4561 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     4621 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     4681 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     4741 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     4801 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttcag gtggcacttt
     4861 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     4921 tccgctcatg aattaattct tagaaaaact catcgagcat caaatgaaac tgcaatttat
     4981 tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa
     5041 actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc
     5101 gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga
     5161 aatcaccatg agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc
     5221 agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac
     5281 cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac
     5341 aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat
     5401 tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg gggatcgcag
     5461 tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca
     5521 taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac
     5581 ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg
     5641 tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca
     5701 tgttggaatt taatcgcggc ctagagcaag acgtttcccg ttgaatatgg ctcataacac
     5761 cccttgtatt actgtttatg taagcagaca gttttattgt tcatgaccaa aatcccttaa
     5821 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa aga
//
