LOCUS       sequence-164916-        5755 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  csd5, construct: 41-192, pET28b(+) N-tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5755)
  AUTHORS   Kim HS, Kim J, Im HN, An DR, Lee M, Hesek D, Mobashery S, Kim JY, 
            Cho K, Yoon HJ, Han BW, Lee BI, Suh SW
  TITLE     Structural basis for the recognition of muramyltripeptide by 
            Helicobacter pylori Csd4, a D,L-carboxypeptidase controlling the 
            helical cell shape.
  JOURNAL   Acta Crystallogr D Biol Crystallogr. 2014 Nov;70(Pt 11):2800-12. 
            doi: 10.1107/S1399004714018732. Epub 2014 Oct 16.
  PUBMED    25372672
REFERENCE   2  (bases 1 to 5755)
  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: "Acta 
            Crystallogr D Biol Crystallogr."; date: "2014-11"; volume: "70(Pt 
            11):2800-12. doi"; pages: " 10.1107/S1399004714018732. Epub 2014 Oct
            16"
COMMENT     Sequence Label: csd5_41-192/pET28b(+)
FEATURES             Location/Qualifiers
     source          1..5755
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      77..665
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     566..585
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     819..836
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    851..993
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(919..938)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1079..1101
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1095..1286)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(2095..3177)
                     /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     3139..3158
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3178..3255)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     3461..3480
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     3564..3583
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3564..3582
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3583..3607
                     /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             3622..3644
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3663..3680
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3690..3707
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             4179..4196
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4249..4267)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4263..4310
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      4347..4802
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4434..4453)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4644..4665
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4895..5710)
                     /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(4895..5710)
                     /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     5618..5637
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5702..5721
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
       61 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
      121 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
      181 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
      241 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
      301 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
      361 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
      421 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
      481 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
      541 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
      601 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
      661 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
      721 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
      781 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
      841 cggaagagcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
      901 tatatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agtatacact
      961 ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac acccgctgac
     1021 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     1081 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg
     1141 taaagctcat cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc
     1201 agctcgttga gtttctccag aagcgttaat gtctggcttc tgataaagcg ggccatgtta
     1261 agggcggttt tttcctgttt ggtcactgat gcctccgtgt aagggggatt tctgttcatg
     1321 ggggtaatga taccgatgaa acgagagagg atgctcacga tacgggttac tgatgatgaa
     1381 catgcccggt tactggaacg ttgtgagggt aaacaactgg cggtatggat gcggcgggac
     1441 cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta atacagatgt aggtgttcca
     1501 cagggtagcc agcagcatcc tgcgatgcag atccggaaca taatggtgca gggcgctgac
     1561 ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag
     1621 gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc
     1681 tgctaaccag taaggcaacc ccgccagcct agccgggtcc tcaacgacag gagcacgatc
     1741 atgcgcaccc gtggggccgc catgccggcg ataatggcct gcttctcgcc gaaacgtttg
     1801 gtggcgggac cagtgacgaa ggcttgagcg agggcgtgca agattccgaa taccgcaagc
     1861 gacaggccga tcatcgtcgc gctccagcga aagcggtcct cgccgaaaat gacccagagc
     1921 gctgccggca cctgtcctac gagttgcatg ataaagaaga cagtcataag tgcggcgacg
     1981 atagtcatgc cccgcgccca ccggaaggag ctgactgggt tgaaggctct caagggcatc
     2041 ggtcgagatc ccggtgccta atgagtgagc taacttacat taattgcgtt gcgctcactg
     2101 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     2161 gggagaggcg gtttgcgtat tgggcgccag ggtggttttt cttttcacca gtgagacggg
     2221 caacagctga ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct
     2281 ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt aacggcggga tataacatga
     2341 gctgtcttcg gtatcgtcgt atcccactac cgagatatcc gcaccaacgc gcagcccgga
     2401 ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg ttggcaacca gcatcgcagt
     2461 gggaacgatg ccctcattca gcatttgcat ggtttgttga aaaccggaca tggcactcca
     2521 gtcgccttcc cgttccgcta tcggctgaat ttgattgcga gtgagatatt tatgccagcc
     2581 agccagacgc agacgcgccg agacagaact taatgggccc gctaacagcg cgatttgctg
     2641 gtgacccaat gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat
     2701 aatactgttg atgggtgtct ggtcagagac atcaagaaat aacgccggaa cattagtgca
     2761 ggcagcttcc acagcaatgg catcctggtc atccagcgga tagttaatga tcagcccact
     2821 gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc
     2881 taccatcgac accaccacgc tggcacccag ttgatcggcg cgagatttaa tcgccgcgac
     2941 aatttgcgac ggcgcgtgca gggccagact ggaggtggca acgccaatca gcaacgactg
     3001 tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc
     3061 ttccactttt tcccgcgttt tcgcagaaac gtggctggcc tggttcacca cgcgggaaac
     3121 ggtctgataa gagacaccgg catactctgc gacatcgtat aacgttactg gtttcacatt
     3181 caccaccctg aattgactct cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg
     3241 ccattcgatg gtgtccggga tctcgacgct ctcccttatg cgactcctgc attaggaagc
     3301 agcccagtag taggttgagg ccgttgagca ccgccgccgc aaggaatggt gcatgcaagg
     3361 agatggcgcc caacagtccc ccggccacgg ggcctgccac catacccacg ccgaaacaag
     3421 cgctcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg gcgatatagg
     3481 cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg gcgtagagga
     3541 tcgagatctc gatcccgcga aattaatacg actcactata ggggaattgt gagcggataa
     3601 caattcccct ctagaaataa ttttgtttaa ctttaagaag gagatatacc atgggcagca
     3661 gccatcatca tcatcatcac agcagcggcc tggtgccgcg cggcagccat atgagcgctc
     3721 caataagccc aaatgtagaa aaaaccgaga cagagcgtca aaacggcgtg ctttcgccca
     3781 aacaagaaga agccaacgca accacaactg ccacagaaga aagccccacc aaagacacag
     3841 cgccgccttt agacacagcc gcgcaaaaac aagaaactaa acaagagcaa gaaaaagaaa
     3901 acgagcctaa acaagatagc gtcccgcccg ttcaaaacaa tcaaaaaacc cctacaaccc
     3961 ccttaatggg aaaaaaacct ttagagtata aagtcgcagt cagtggcgtg aatgtgcgcg
     4021 cttttcccag cacaaaaggt aaaatcttgg gattgctttt aaaaaataaa agcgtgaaag
     4081 ttttagaaat ccaaaacgat tgggctgaaa ttgaattttc tcacgaaaca aagggctatg
     4141 tgtttttaaa acttttaaaa aaggctgaat aactcgagca ccaccaccac caccactgag
     4201 atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat
     4261 aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag
     4321 gaactatatc cggattggcg aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg
     4381 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     4441 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     4501 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     4561 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     4621 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     4681 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     4741 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat
     4801 ttcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     4861 acattcaaat atgtatccgc tcatgaatta attcttagaa aaactcatcg agcatcaaat
     4921 gaaactgcaa tttattcata tcaggattat caataccata tttttgaaaa agccgtttct
     4981 gtaatgaagg agaaaactca ccgaggcagt tccataggat ggcaagatcc tggtatcggt
     5041 ctgcgattcc gactcgtcca acatcaatac aacctattaa tttcccctcg tcaaaaataa
     5101 ggttatcaag tgagaaatca ccatgagtga cgactgaatc cggtgagaat ggcaaaagtt
     5161 tatgcatttc tttccagact tgttcaacag gccagccatt acgctcgtca tcaaaatcac
     5221 tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg agcgagacga aatacgcgat
     5281 cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg aacactgcca
     5341 gcgcatcaac aatattttca cctgaatcag gatattcttc taatacctgg aatgctgttt
     5401 tcccggggat cgcagtggtg agtaaccatg catcatcagg agtacggata aaatgcttga
     5461 tggtcggaag aggcataaat tccgtcagcc agtttagtct gaccatctca tctgtaacat
     5521 cattggcaac gctacctttg ccatgtttca gaaacaactc tggcgcatcg ggcttcccat
     5581 acaatcgata gattgtcgca cctgattgcc cgacattatc gcgagcccat ttatacccat
     5641 ataaatcagc atccatgttg gaatttaatc gcggcctaga gcaagacgtt tcccgttgaa
     5701 tatggctcat aacacccctt gtattactgt ttatgtaagc agacagtttt attgt
//
