LOCUS       sequence-9588-ex        5701 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 5701)
  AUTHORS   Hu H, Clarkson MW, Hermans J, Lee AL
  TITLE     Increased rigidity of eglin c at acidic pH: evidence from NMR spin 
            relaxation and MD simulations.
  JOURNAL   Biochemistry. 2003 Dec 2. 42(47):13856-68.
  PUBMED    14636053
REFERENCE   2  (bases 1 to 5701)
  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: 
            "Biochemistry. 2003 Dec 2. 42(47):13856-68."
COMMENT     Sequence Label: pET21a- hPTP1e-PDZ2
FEATURES             Location/Qualifiers
     source          1..5701
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(26..73)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     69..87
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(140..157)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             505..527
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    542..566
                     /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)."
     primer_bind     complement(566..585)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(567..585)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(669..688)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        894..971
                     /gene="lacI"
                     /label=lacI promoter
     CDS             972..2054
                     /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     complement(991..1010)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             2863..3054
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(3048..3070)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3156..3298
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3211..3230
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3313..3330)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3484..4072)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3564..3583)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4243..5034,5035..5103))
                     /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     4866..4885
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5104..5208)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5235..5690)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5372..5393)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5584..5603
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
       61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
      121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagtgcggcc gcaagcttgt
      181 cgacggagct cgaattcgga tccttatgtt ggagattgtc ccttttctaa taacagatga
      241 accacctgtc ctgtatttct cagtgtttcc acagcttgct tatgggtggc tccttctaga
      301 ctaactccat tgacagctag gacgcgatca cctttgtgaa ttctaccatc agactctgct
      361 gctccctggg gaataacagc tttcacataa atgccaccat gtctgacact cgtattcaca
      421 cctcccgtga cacttatccc caagctgtta tcatttttag ccagttcaac ctcaaagata
      481 tctccaggct taggcatatg tatatctcct tcttaaagtt aaacaaaatt atttctagag
      541 gggaattgtt atccgctcac aattccccta tagtgagtcg tattaatttc gcgggatcga
      601 gatctcgatc ctctacgccg gacgcatcgt ggccggcatc accggcgcca caggtgcggt
      661 tgctggcgcc tatatcgccg acatcaccga tggggaagat cgggctcgcc acttcgggct
      721 catgagcgct tgtttcggcg tgggtatggt ggcaggcccc gtggccgggg gactgttggg
      781 cgccatctcc ttgcatgcac cattccttgc ggcggcggtg ctcaacggcc tcaacctact
      841 actgggctgc ttcctaatgc aggagtcgca taagggagag cgtcgagatc ccggacacca
      901 tcgaatggcg caaaaccttt cgcggtatgg catgatagcg cccggaagag agtcaattca
      961 gggtggtgaa tgtgaaacca gtaacgttat acgatgtcgc agagtatgcc ggtgtctctt
     1021 atcagaccgt ttcccgcgtg gtgaaccagg ccagccacgt ttctgcgaaa acgcgggaaa
     1081 aagtggaagc ggcgatggcg gagctgaatt acattcccaa ccgcgtggca caacaactgg
     1141 cgggcaaaca gtcgttgctg attggcgttg ccacctccag tctggccctg cacgcgccgt
     1201 cgcaaattgt cgcggcgatt aaatctcgcg ccgatcaact gggtgccagc gtggtggtgt
     1261 cgatggtaga acgaagcggc gtcgaagcct gtaaagcggc ggtgcacaat cttctcgcgc
     1321 aacgcgtcag tgggctgatc attaactatc cgctggatga ccaggatgcc attgctgtgg
     1381 aagctgcctg cactaatgtt ccggcgttat ttcttgatgt ctctgaccag acacccatca
     1441 acagtattat tttctcccat gaagacggta cgcgactggg cgtggagcat ctggtcgcat
     1501 tgggtcacca gcaaatcgcg ctgttagcgg gcccattaag ttctgtctcg gcgcgtctgc
     1561 gtctggctgg ctggcataaa tatctcactc gcaatcaaat tcagccgata gcggaacggg
     1621 aaggcgactg gagtgccatg tccggttttc aacaaaccat gcaaatgctg aatgagggca
     1681 tcgttcccac tgcgatgctg gttgccaacg atcagatggc gctgggcgca atgcgcgcca
     1741 ttaccgagtc cgggctgcgc gttggtgcgg atatctcggt agtgggatac gacgataccg
     1801 aagacagctc atgttatatc ccgccgttaa ccaccatcaa acaggatttt cgcctgctgg
     1861 ggcaaaccag cgtggaccgc ttgctgcaac tctctcaggg ccaggcggtg aagggcaatc
     1921 agctgttgcc cgtctcactg gtgaaaagaa aaaccaccct ggcgcccaat acgcaaaccg
     1981 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     2041 aaagcgggca gtgagcgcaa cgcaattaat gtaagttagc tcactcatta ggcaccggga
     2101 tctcgaccga tgcccttgag agccttcaac ccagtcagct ccttccggtg ggcgcggggc
     2161 atgactatcg tcgccgcact tatgactgtc ttctttatca tgcaactcgt aggacaggtg
     2221 ccggcagcgc tctgggtcat tttcggcgag gaccgctttc gctggagcgc gacgatgatc
     2281 ggcctgtcgc ttgcggtatt cggaatcttg cacgccctcg ctcaagcctt cgtcactggt
     2341 cccgccacca aacgtttcgg cgagaagcag gccattatcg ccggcatggc ggccccacgg
     2401 gtgcgcatga tcgtgctcct gtcgttgagg acccggctag gctggcgggg ttgccttact
     2461 ggttagcaga atgaatcacc gatacgcgag cgaacgtgaa gcgactgctg ctgcaaaacg
     2521 tctgcgacct gagcaacaac atgaatggtc ttcggtttcc gtgtttcgta aagtctggaa
     2581 acgcggaagt cagcgccctg caccattatg ttccggatct gcatcgcagg atgctgctgg
     2641 ctaccctgtg gaacacctac atctgtatta acgaagcgct ggcattgacc ctgagtgatt
     2701 tttctctggt cccgccgcat ccataccgcc agttgtttac cctcacaacg ttccagtaac
     2761 cgggcatgtt catcatcagt aacccgtatc gtgagcatcc tctctcgttt catcggtatc
     2821 attaccccca tgaacagaaa tcccccttac acggaggcat cagtgaccaa acaggaaaaa
     2881 accgccctta acatggcccg ctttatcaga agccagacat taacgcttct ggagaaactc
     2941 aacgagctgg acgcggatga acaggcagac atctgtgaat cgcttcacga ccacgctgat
     3001 gagctttacc gcagctgcct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
     3061 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt
     3121 cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc
     3181 gatagcggag tgtatactgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
     3241 accatatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
     3301 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     3361 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     3421 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     3481 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     3541 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     3601 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     3661 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     3721 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     3781 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     3841 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     3901 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     3961 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     4021 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     4081 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     4141 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     4201 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     4261 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     4321 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     4381 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     4441 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     4501 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctgc
     4561 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     4621 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     4681 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     4741 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     4801 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     4861 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     4921 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     4981 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     5041 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     5101 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     5161 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     5221 aaaagtgcca cctgaaattg taaacgttaa tattttgtta aaattcgcgt taaatttttg
     5281 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     5341 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     5401 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     5461 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     5521 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     5581 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
     5641 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt cccattcgcc
     5701 a
//
