LOCUS       sequence-199731-        6674 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 6674)
  AUTHORS   Yaffe MB, Schutkowski M, Shen M, Zhou XZ, Stukenberg PT, Rahfeld JU,
            Xu J, Kuang J, Kirschner MW, Fischer G, Cantley LC, Lu KP
  TITLE     Sequence-specific and phosphorylation-dependent proline 
            isomerization: a potential mitotic regulatory mechanism.
  JOURNAL   Science. 1997 Dec 12. 278(5345):1957-60.
  PUBMED    9395400
REFERENCE   2  (bases 1 to 6674)
  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: "Science. 
            1997 Dec 12. 278(5345):1957-60."
COMMENT     Sequence Label: GST-Pin1
FEATURES             Location/Qualifiers
     source          1..6674
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(125..130,131..146,147..153)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    161..177
                     /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)."
     CDS             200..853
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     811..833
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             860..877
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(1959..1978)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1960..1978)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1985..2001)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2666..2688)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2833..2851)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2919..3023
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3024..3092,3093..3884)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3242..3261)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4055..4643
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4544..4563
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4797..4814
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4887..4964
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4965..6047
                     /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(4984..5003)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(6096..6101,6102..6119,6120..6126)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6134..6150
                     /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     6158..6174
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6158..6174
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(6170..6674,1..23)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(6190..6206)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tggtatggct gtgcaggtcg taaatcactg cataattcgt gtcgctcaag gcgcactccc
       61 gttctggata atgttttttg cgccgacatc ataacggttc tggcaaatat tctgaaatga
      121 gctgttgaca attaatcatc ggctcgtata atgtgtggaa ttgtgagcgg ataacaattt
      181 cacacaggaa acagtattca tgtcccctat actaggttat tggaaaatta agggccttgt
      241 gcaacccact cgacttcttt tggaatatct tgaagaaaaa tatgaagagc atttgtatga
      301 gcgcgatgaa ggtgataaat ggcgaaacaa aaagtttgaa ttgggtttgg agtttcccaa
      361 tcttccttat tatattgatg gtgatgttaa attaacacag tctatggcca tcatacgtta
      421 tatagctgac aagcacaaca tgttgggtgg ttgtccaaaa gagcgtgcag agatttcaat
      481 gcttgaagga gcggttttgg atattagata cggtgtttcg agaattgcat atagtaaaga
      541 ctttgaaact ctcaaagttg attttcttag caagctacct gaaatgctga aaatgttcga
      601 agatcgttta tgtcataaaa catatttaaa tggtgatcat gtaacccatc ctgacttcat
      661 gttgtatgac gctcttgatg ttgttttata catggaccca atgtgcctgg atgcgttccc
      721 aaaattagtt tgttttaaaa aacgtattga agctatccca caaattgata agtacttgaa
      781 atccagcaag tatatagcat ggcctttgca gggctggcaa gccacgtttg gtggtggcga
      841 ccatcctcca aaatcggatc tggttccgcg tggatccccg ggaatttccg gtggtggtgg
      901 tggaattcta gactccatga attcggcacg agggaagatg gcggacgagg agaagctgcc
      961 gcccggctgg gagaagcgca tgagccgcag ctcaggccga gtgtactact tcaaccacat
     1021 cactaacgcc agccagtggg agcggcccag cggcaacagc agcagtggtg gcaaaaacgg
     1081 gcagggggag cctgccaggg tccgctgctc gcacctgctg gtgaagcaca gccagtcacg
     1141 gcggccctcg tcctggcggc aggagaagat cacccggacc aaggaggagg ccctggagct
     1201 gatcaacggc tacatccaga agatcaagtc gggagaggag gactttgagt ctctggcctc
     1261 acagttcagc gactgcagct cagccaaggc caggggagac ctgggtgcct tcagcagagg
     1321 tcagatgcag aagccatttg aagacgcctc gtttgcgctg cggacggggg agatgagcgg
     1381 gcccgtgttc acggattccg gcatccacat catcctccgc actgagtgag ggtggggagc
     1441 ccaggcctgg cctcggggca gggcagggcg gctaggccgg ccagctcccc cttgcccgcc
     1501 agccagtggc cgaacccccc actccctgcc accgtcacac agtatttatt gttcccacaa
     1561 tggctgggag ggggcccttc cagattgggg gccctggggt ccccactccc tgtccatccc
     1621 cagttggggc tgcgaccgcc agattctccc ttaaggaatt gacttcagca ggggtgggag
     1681 gctcccagac ccagggcagt gtggtgggag gggtgttcca aagagaaggc ctggtcagca
     1741 gagccgcccc gtgtcccccc aggtgctgga ggcagactcg agggccgaat tgtttctagt
     1801 taggccacgc tcctctgttc agtcgcaaag gtgaacactc atgcggccca gccatgggcc
     1861 ctctgagcaa ctgtgcagca ccctttcacc cccaattaaa cccagaacca ctaaaaaaaa
     1921 aaaaaaaaaa ctcgaggggg ggcccggtac ccaattcgcc ctatagtgag tcgtattaca
     1981 attcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctgatcta tgaatcgtag
     2041 atactgaaaa accccgcaag ttcacttcaa ctgtgcatcg tgcaccatct caatttcttt
     2101 catttataca tcgttttgcc ttcttttatg taactatact cctctaagtt tcaatcttgg
     2161 ccatgtaacc tctgatctat agaatttttt aaatgactag aattaatgcc catctttttt
     2221 ttggacctaa attcttcatg aaaatatatt acgagggctt attcagaagc tacaacaaca
     2281 attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt
     2341 aaaacctcta caaatgtggt atggctgatt atgatcctgc aagcctcgtc gtcctggccg
     2401 gaccacgcta tctgtgcaag gtccccggcc ccggacgcgc gctccatgag cagagcgccc
     2461 gccgccgagg cgaagactcg ggcggcgccc tgcccgtccc accaggtcaa caggcggtaa
     2521 ccggcctctt catcgggaat gcgcgcgacc ttcagcatcg ccggcatgtc cccctggcgg
     2581 acgggaagta tccagctcga ccaagcttaa ttcatcgtga ctgactgacg atctgcctcg
     2641 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
     2701 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     2761 gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga tagcggagtg tataattctt
     2821 gaagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
     2881 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     2941 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     3001 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     3061 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     3121 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     3181 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     3241 tgctatgtgg cgcggtatta tcccgtgttg acgccgggca agagcaactc ggtcgccgca
     3301 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     3361 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     3421 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     3481 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     3541 acgacgagcg tgacaccacg atgcctgcag caatggcaac aacgttgcgc aaactattaa
     3601 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     3661 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     3721 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     3781 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     3841 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     3901 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     3961 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     4021 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     4081 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     4141 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     4201 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     4261 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     4321 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     4381 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     4441 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     4501 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     4561 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     4621 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     4681 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     4741 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     4801 agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt
     4861 gcggtatttc acaccgcata aattccgaca ccatcgaatg gcgcaaaacc tttcgcggta
     4921 tggcatgata gcgcccggaa gagagtcaat tcagggtggt gaatgtgaaa ccagtaacgt
     4981 tatacgatgt cgcagagtat gccggtgtct cttatcagac cgtttcccgc gtggtgaacc
     5041 aggccagcca cgtttctgcg aaaacgcggg aaaaagtgga agcggcgatg gcggagctga
     5101 attacattcc caaccgcgtg gcacaacaac tggcgggcaa acagtcgttg ctgattggcg
     5161 ttgccacctc cagtctggcc ctgcacgcgc cgtcgcaaat tgtcgcggcg attaaatctc
     5221 gcgccgatca actgggtgcc agcgtggtgg tgtcgatggt agaacgaagc ggcgtcgaag
     5281 cctgtaaagc ggcggtgcac aatcttctcg cgcaacgcgt cagtgggctg atcattaact
     5341 atccgctgga tgaccaggat gccattgctg tggaagctgc ctgcactaat gttccggcgt
     5401 tatttcttga tgtctctgac cagacaccca tcaacagtat tattttctcc catgaagacg
     5461 gtacgcgact gggcgtggag catctggtcg cattgggtca ccagcaaatc gcgctgttag
     5521 cgggcccatt aagttctgtc tcggcgcgtc tgcgtctggc tggctggcat aaatatctca
     5581 ctcgcaatca aattcagccg atagcggaac gggaaggcga ctggagtgcc atgtccggtt
     5641 ttcaacaaac catgcaaatg ctgaatgagg gcatcgttcc cactgcgatg ctggttgcca
     5701 acgatcagat ggcgctgggc gcaatgcgcg ccattaccga gtccgggctg cgcgttggtg
     5761 cggatatctc ggtagtggga tacgacgata ccgaagacag ctcatgttat atcccgccgt
     5821 taaccaccat caaacaggat tttcgcctgc tggggcaaac cagcgtggac cgcttgctgc
     5881 aactctctca gggccaggcg gtgaagggca atcagctgtt gcccgtctca ctggtgaaaa
     5941 gaaaaaccac cctggcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     6001 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     6061 aatgtaagtt agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt
     6121 atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat
     6181 tacggattca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca
     6241 acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg
     6301 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggcgct ttgcctggtt
     6361 tccggcacca gaagcggtgc cggaaagctg gctggagtgc gatcttcctg aggccgatac
     6421 tgtcgtcgtc ccctcaaact ggcagatgca cggttacgat gcgcccatct acaccaacgt
     6481 gacctatccc attacggtca atccgccgtt tgttcccacg gagaatccga cgggttgtta
     6541 ctcgctcaca tttaatgttg atgaaagctg gctacaggaa ggccagacgc gaattatttt
     6601 tgatggcgtt ggaattagct tatcgactgc acggtgcacc aatgcttctg gcgtcaggca
     6661 gccatcggaa gctg
//
