LOCUS       sequence-164865-        6129 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses m23 peptidase from Campylobacter jejuni.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6129)
  TITLE     C.jejuni_peptidase m23 family
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6129)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pET21a(+)_C.jejuni m23 peptidase
FEATURES             Location/Qualifiers
     source          1..6129
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      70..658
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     559..578
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     812..829
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    844..986
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(912..931)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1072..1094
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1088..1279)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(2088..3170)
                     /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     3132..3151
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3171..3248)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     3454..3473
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     3557..3576
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3557..3575
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3576..3600
                     /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             3615..3637
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4413..4430
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4483..4501)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4497..4544
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      4581..5036
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4668..4687)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4878..4899
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5063..5167
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5168..5236,5237..6028)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5386..5405)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
       61 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
      121 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
      181 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
      241 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
      301 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
      361 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
      421 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
      481 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
      541 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
      601 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
      661 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
      721 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
      781 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      841 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatatgg
      901 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat
      961 cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct
     1021 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     1081 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct
     1141 catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt
     1201 tgagtttctc cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg
     1261 ttttttcctg tttggtcact gatgcctccg tgtaaggggg atttctgttc atgggggtaa
     1321 tgataccgat gaaacgagag aggatgctca cgatacgggt tactgatgat gaacatgccc
     1381 ggttactgga acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa
     1441 aaatcactca gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta
     1501 gccagcagca tcctgcgatg cagatccgga acataatggt gcagggcgct gacttccgcg
     1561 tttccagact ttacgaaaca cggaaaccga agaccattca tgttgttgct caggtcgcag
     1621 acgttttgca gcagcagtcg cttcacgttc gctcgcgtat cggtgattca ttctgctaac
     1681 cagtaaggca accccgccag cctagccggg tcctcaacga caggagcacg atcatgcgca
     1741 cccgtggggc cgccatgccg gcgataatgg cctgcttctc gccgaaacgt ttggtggcgg
     1801 gaccagtgac gaaggcttga gcgagggcgt gcaagattcc gaataccgca agcgacaggc
     1861 cgatcatcgt cgcgctccag cgaaagcggt cctcgccgaa aatgacccag agcgctgccg
     1921 gcacctgtcc tacgagttgc atgataaaga agacagtcat aagtgcggcg acgatagtca
     1981 tgccccgcgc ccaccggaag gagctgactg ggttgaaggc tctcaagggc atcggtcgag
     2041 atcccggtgc ctaatgagtg agctaactta cattaattgc gttgcgctca ctgcccgctt
     2101 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     2161 gcggtttgcg tattgggcgc cagggtggtt tttcttttca ccagtgagac gggcaacagc
     2221 tgattgccct tcaccgcctg gccctgagag agttgcagca agcggtccac gctggtttgc
     2281 cccagcaggc gaaaatcctg tttgatggtg gttaacggcg ggatataaca tgagctgtct
     2341 tcggtatcgt cgtatcccac taccgagata tccgcaccaa cgcgcagccc ggactcggta
     2401 atggcgcgca ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc agtgggaacg
     2461 atgccctcat tcagcatttg catggtttgt tgaaaaccgg acatggcact ccagtcgcct
     2521 tcccgttccg ctatcggctg aatttgattg cgagtgagat atttatgcca gccagccaga
     2581 cgcagacgcg ccgagacaga acttaatggg cccgctaaca gcgcgatttg ctggtgaccc
     2641 aatgcgacca gatgctccac gcccagtcgc gtaccgtctt catgggagaa aataatactg
     2701 ttgatgggtg tctggtcaga gacatcaaga aataacgccg gaacattagt gcaggcagct
     2761 tccacagcaa tggcatcctg gtcatccagc ggatagttaa tgatcagccc actgacgcgt
     2821 tgcgcgagaa gattgtgcac cgccgcttta caggcttcga cgccgcttcg ttctaccatc
     2881 gacaccacca cgctggcacc cagttgatcg gcgcgagatt taatcgccgc gacaatttgc
     2941 gacggcgcgt gcagggccag actggaggtg gcaacgccaa tcagcaacga ctgtttgccc
     3001 gccagttgtt gtgccacgcg gttgggaatg taattcagct ccgccatcgc cgcttccact
     3061 ttttcccgcg ttttcgcaga aacgtggctg gcctggttca ccacgcggga aacggtctga
     3121 taagagacac cggcatactc tgcgacatcg tataacgtta ctggtttcac attcaccacc
     3181 ctgaattgac tctcttccgg gcgctatcat gccataccgc gaaaggtttt gcgccattcg
     3241 atggtgtccg ggatctcgac gctctccctt atgcgactcc tgcattagga agcagcccag
     3301 tagtaggttg aggccgttga gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc
     3361 gcccaacagt cccccggcca cggggcctgc caccataccc acgccgaaac aagcgctcat
     3421 gagcccgaag tggcgagccc gatcttcccc atcggtgatg tcggcgatat aggcgccagc
     3481 aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga ggatcgagat
     3541 ctcgatcccg cgaaattaat acgactcact ataggggaat tgtgagcgga taacaattcc
     3601 cctctagaaa taattttgtt taactttaag aaggagatat acatatggaa ctcatcaaag
     3661 gtcaagcttt atttttggaa ctagataaga aagatttttt atctttgaaa aataatgata
     3721 aaaatattcc cacctttgca catcctaaaa atcaagaaaa aattctagct attttttcct
     3781 taccttacaa aaatcccccg caaaatacaa aacttatcgc attttataaa gataaaaaag
     3841 aagaaatttt cataaaaact ttagaaggta attataaaag tgaaaaattg caagttgaaa
     3901 acaaaaagat ttttccacca aaaaccatac aagaacgcat cgctaaagaa ttaaaagaag
     3961 ccaatgcgat ttatagttct tatactccaa aagctttatt taacggtgct tttaatatac
     4021 ctttaaattc ttttatcaca agtgattttg gcaaagcaag aacttttaat gaaaaagtag
     4081 caagttatca tagtggaacg gactttagag ccgccacagg aacgcctatt tatgcagcca
     4141 attcaggtgt agtaaaaatt gcaaaagatc gttatttcgc aggaaattca gtagtcattg
     4201 atcatggttt tggaatttat tcacaatatt atcatctttc taaaatcgat gttaaagtag
     4261 gacaaaagat aaaaaaaggt gaacttatag gacttagtgg ggctagtggt agggtaagtg
     4321 ggccgcattt gcattttgga attttagctg gaggcaaaca agttgatcct ttggattttg
     4381 tatccaaatt taatgctatt tttcaactcg agcaccacca ccaccaccac tgagatccgg
     4441 ctgctaacaa agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactag
     4501 cataacccct tggggcctct aaacgggtct tgaggggttt tttgctgaaa ggaggaacta
     4561 tatccggatt ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     4621 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     4681 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     4741 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     4801 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     4861 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     4921 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     4981 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta caatttcagg
     5041 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     5101 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     5161 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     5221 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     5281 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     5341 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     5401 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     5461 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     5521 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     5581 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     5641 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     5701 cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     5761 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     5821 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     5881 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     5941 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     6001 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     6061 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     6121 tctcatgac
//
