LOCUS       sequence-48333-e        6111 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6111)
  AUTHORS   Shen A, Lupardus PJ, Morell M, Ponder EL, Sadaghiani AM, Garcia KC, 
            Bogyo M
  TITLE     Simplified, enhanced protein purification using an inducible, 
            autoprocessing enzyme tag.
  JOURNAL   PLoS One. 2009 Dec 2;4(12):e8119.
  PUBMED    19956581
REFERENCE   2  (bases 1 to 6111)
  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: "2009-12-2"; volume: "4(12)"; pages: "e8119"
COMMENT     Sequence Label: pET22b-CPDSalI
FEATURES             Location/Qualifiers
     source          1..6111
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      18..473
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(105..124)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     315..336
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        500..604
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(605..673,674..1465)
                     /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(823..842)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1636..2224
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2125..2144
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2378..2395
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2410..2552
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2478..2497)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2638..2660
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2654..2845)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3654..4736)
                     /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     4698..4717
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4737..4814)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5020..5039
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5123..5142
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5123..5141
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5142..5166
                     /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             5181..5203
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     sig_peptide     5211..5276
                     /codon_start=1
                     /gene="pelB (fragment)"
                     /product="leader peptide for secretion"
                     /label=pelB signal sequence
                     /translation="MKYLLPTAAAGLLLLAAQPAMA"
     CDS             5961..5978
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6031..6049)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      6045..6092
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 ccggattggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
       61 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
      121 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
      181 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
      241 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
      301 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
      361 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
      421 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa tttcaggtgg
      481 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
      541 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
      601 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
      661 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
      721 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
      781 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
      841 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
      901 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
      961 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     1021 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     1081 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     1141 gatgcctgca gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     1201 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     1261 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     1321 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     1381 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     1441 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     1501 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     1561 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     1621 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     1681 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     1741 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta
     1801 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     1861 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     1921 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     1981 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     2041 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     2101 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     2161 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     2221 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     2281 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     2341 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     2401 ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     2461 atatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc
     2521 cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg
     2581 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     2641 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt
     2701 aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca
     2761 gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa
     2821 gggcggtttt ttcctgtttg gtcactgatg cctccgtgta agggggattt ctgttcatgg
     2881 gggtaatgat accgatgaaa cgagagagga tgctcacgat acgggttact gatgatgaac
     2941 atgcccggtt actggaacgt tgtgagggta aacaactggc ggtatggatg cggcgggacc
     3001 agagaaaaat cactcagggt caatgccagc gcttcgttaa tacagatgta ggtgttccac
     3061 agggtagcca gcagcatcct gcgatgcaga tccggaacat aatggtgcag ggcgctgact
     3121 tccgcgtttc cagactttac gaaacacgga aaccgaagac cattcatgtt gttgctcagg
     3181 tcgcagacgt tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt gattcattct
     3241 gctaaccagt aaggcaaccc cgccagccta gccgggtcct caacgacagg agcacgatca
     3301 tgcgcacccg tggggccgcc atgccggcga taatggcctg cttctcgccg aaacgtttgg
     3361 tggcgggacc agtgacgaag gcttgagcga gggcgtgcaa gattccgaat accgcaagcg
     3421 acaggccgat catcgtcgcg ctccagcgaa agcggtcctc gccgaaaatg acccagagcg
     3481 ctgccggcac ctgtcctacg agttgcatga taaagaagac agtcataagt gcggcgacga
     3541 tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt gaaggctctc aagggcatcg
     3601 gtcgagatcc cggtgcctaa tgagtgagct aacttacatt aattgcgttg cgctcactgc
     3661 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     3721 ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc ttttcaccag tgagacgggc
     3781 aacagctgat tgcccttcac cgcctggccc tgagagagtt gcagcaagcg gtccacgctg
     3841 gtttgcccca gcaggcgaaa atcctgtttg atggtggtta acggcgggat ataacatgag
     3901 ctgtcttcgg tatcgtcgta tcccactacc gagatatccg caccaacgcg cagcccggac
     3961 tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag catcgcagtg
     4021 ggaacgatgc cctcattcag catttgcatg gtttgttgaa aaccggacat ggcactccag
     4081 tcgccttccc gttccgctat cggctgaatt tgattgcgag tgagatattt atgccagcca
     4141 gccagacgca gacgcgccga gacagaactt aatgggcccg ctaacagcgc gatttgctgg
     4201 tgacccaatg cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata
     4261 atactgttga tgggtgtctg gtcagagaca tcaagaaata acgccggaac attagtgcag
     4321 gcagcttcca cagcaatggc atcctggtca tccagcggat agttaatgat cagcccactg
     4381 acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct
     4441 accatcgaca ccaccacgct ggcacccagt tgatcggcgc gagatttaat cgccgcgaca
     4501 atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag caacgactgt
     4561 ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc catcgccgct
     4621 tccacttttt cccgcgtttt cgcagaaacg tggctggcct ggttcaccac gcgggaaacg
     4681 gtctgataag agacaccggc atactctgcg acatcgtata acgttactgg tttcacattc
     4741 accaccctga attgactctc ttccgggcgc tatcatgcca taccgcgaaa ggttttgcgc
     4801 cattcgatgg tgtccgggat ctcgacgctc tcccttatgc gactcctgca ttaggaagca
     4861 gcccagtagt aggttgaggc cgttgagcac cgccgccgca aggaatggtg catgcaagga
     4921 gatggcgccc aacagtcccc cggccacggg gcctgccacc atacccacgc cgaaacaagc
     4981 gctcatgagc ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg cgatataggc
     5041 gccagcaacc gcacctgtgg cgccggtgat gccggccacg atgcgtccgg cgtagaggat
     5101 cgagatctcg atcccgcgaa attaatacga ctcactatag gggaattgtg agcggataac
     5161 aattcccctc tagaaataat tttgtttaac tttaagaagg agatatacat atgaaatacc
     5221 tgctgccgac cgctgctgct ggtctgctgc tcctcgctgc ccagccggcg atggccatgg
     5281 atatcggaat taattcggat ccgaattcga gctccgtcga cgcattagcg gatggaaaaa
     5341 tactccataa tcaaaatgtt aatagctggg gcccgattac ggttacacca acgacagatg
     5401 gtggtgaaac ccgcttcgac ggtcaaatca tcgttcaaat ggaaaacgac ccggtagtag
     5461 caaaagcggc agccaattta gcaggtaaac atgctgaaag cagtgtggtg gtgcagctcg
     5521 attcagacgg caactatcgc gtggtgtatg gcgatccgtc aaaactggat ggaaagctac
     5581 gttggcagtt ggtggggcat ggtcgcgacc actcagaaac taacaatact cgcttaagtg
     5641 gttacagtgc cgatgagttg gccgtgaaat tggccaagtt ccaacagtcg tttaatcaag
     5701 ccgaaaacat caacaacaaa ccggatcaca tcagtattgt tggttgttct ttggtgagtg
     5761 acgacaagca aaaaggcttt ggtcatcagt ttattaacgc gatggatgcg aatggtcttc
     5821 gtgtcgatgt ctctgttcgt agttctgaac tggccgtaga cgaggcggga cgtaagcata
     5881 ccaaggacgc gaatggcgat tgggttcaaa aggcagaaaa caacaaagtt tcgctaagct
     5941 gggacgcgca aggtctcgag caccaccacc accaccactg agatccggct gctaacaaag
     6001 cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg
     6061 gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata t
//
