LOCUS       sequence-199760-        6219 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  human PTP1B catalytic domain (residues 1-301) in RP1B vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6219)
  AUTHORS   Choy MS, Li Y, Machado LE, Kunze MB, Connors CR, Wei X, 
            Lindorff-Larsen K, Page R, Peti W
  TITLE     Conformational Rigidity and Protein Dynamics at Distinct Timescales 
            Regulate PTP1B Activity and Allostery.
  JOURNAL   Mol Cell. 2017 Feb 16;65(4):644-658.e5. doi: 
            10.1016/j.molcel.2017.01.014.
  PUBMED    28212750
REFERENCE   2  (bases 1 to 6219)
  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"; doi: 
            "10.1016/j.molcel.2017.01"; journalName: "Mol Cell"; date: 
            "2017-02-16- 16"; volume: "65"; issue: "4"; pages: "644-658.e5"
COMMENT     Sequence Label: PTP1B human
FEATURES             Location/Qualifiers
     source          1..6219
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             165..182
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(235..253)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      249..296
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      333..788
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(420..439)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     630..651
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(880..1695)
                     /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(880..1695)
                     /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     1603..1622
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1687..1706
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1817..2405
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2306..2325
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2559..2576
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2591..2730
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2656..2675)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2816..2838
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2832..3023)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3832..4914)
                     /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     4876..4895
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4915..4992)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5198..5217
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5301..5320
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5301..5319
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5320..5344
                     /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             5359..5381
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5406..5423
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             5424..5444
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
ORIGIN
        1 agtttcggat ggggctgatc cagacagccg accagctgcg cttctcctac ctggctgtga
       61 tcgaaggtgc caaattcatc atgggggact cttccgtgca ggatcagtgg aaggagcttt
      121 cccacgagga cctggagcca cacaattgag cggccgcact cgagcaccac caccaccacc
      181 actgagatcc ggctgctaac aaagcccgaa aggaagctga gttggctgct gccaccgctg
      241 agcaataact agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga
      301 aaggaggaac tatatccgga ttggcgaatg ggacgcgccc tgtagcggcg cattaagcgc
      361 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
      421 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
      481 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
      541 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
      601 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
      661 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
      721 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct
      781 tacaatttag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
      841 taaatacatt caaatatgta tccgctcatg aattaattct tagaaaaact catcgagcat
      901 caaatgaaac tgcaatttat tcatatcagg attatcaata ccatattttt gaaaaagccg
      961 tttctgtaat gaaggagaaa actcaccgag gcagttccat aggatggcaa gatcctggta
     1021 tcggtctgcg attccgactc gtccaacatc aatacaacct attaatttcc cctcgtcaaa
     1081 aataaggtta tcaagtgaga aatcaccatg agtgacgact gaatccggtg agaatggcaa
     1141 aagtttatgc atttctttcc agacttgttc aacaggccag ccattacgct cgtcatcaaa
     1201 atcactcgca tcaaccaaac cgttattcat tcgtgattgc gcctgagcga gacgaaatac
     1261 gcgatcgctg ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc gcaggaacac
     1321 tgccagcgca tcaacaatat tttcacctga atcaggatat tcttctaata cctggaatgc
     1381 tgttttcccg gggatcgcag tggtgagtaa ccatgcatca tcaggagtac ggataaaatg
     1441 cttgatggtc ggaagaggca taaattccgt cagccagttt agtctgacca tctcatctgt
     1501 aacatcattg gcaacgctac ctttgccatg tttcagaaac aactctggcg catcgggctt
     1561 cccatacaat cgatagattg tcgcacctga ttgcccgaca ttatcgcgag cccatttata
     1621 cccatataaa tcagcatcca tgttggaatt taatcgcggc ctagagcaag acgtttcccg
     1681 ttgaatatgg ctcataacac cccttgtatt actgtttatg taagcagaca gttttattgt
     1741 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     1801 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     1861 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     1921 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     1981 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     2041 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     2101 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     2161 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     2221 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     2281 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     2341 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     2401 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     2461 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     2521 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     2581 cggaagagcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     2641 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg
     2701 ctatcgctac gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg
     2761 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     2821 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa
     2881 agctcatcag cgtggtcgtg aagcgattca cagatgtctg cctgttcatc cgcgtccagc
     2941 tcgttgagtt tctccagaag cgttaatgtc tggcttctga taaagcgggc catgttaagg
     3001 gcggtttttt cctgtttggt cactgatgcc tccgtgtaag ggggatttct gttcatgggg
     3061 gtaatgatac cgatgaaacg agagaggatg ctcacgatac gggttactga tgatgaacat
     3121 gcccggttac tggaacgttg tgagggtaaa caactggcgg tatggatgcg gcgggaccag
     3181 agaaaaatca ctcagggtca atgccagcgc ttcgttaata cagatgtagg tgttccacag
     3241 ggtagccagc agcatcctgc gatgcagatc cggaacataa tggtgcaggg cgctgacttc
     3301 cgcgtttcca gactttacga aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc
     3361 gcagacgttt tgcagcagca gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc
     3421 taaccagtaa ggcaaccccg ccagcctagc cgggtcctca acgacaggag cacgatcatg
     3481 cgcacccgtg gggccgccat gccggcgata atggcctgct tctcgccgaa acgtttggtg
     3541 gcgggaccag tgacgaaggc ttgagcgagg gcgtgcaaga ttccgaatac cgcaagcgac
     3601 aggccgatca tcgtcgcgct ccagcgaaag cggtcctcgc cgaaaatgac ccagagcgct
     3661 gccggcacct gtcctacgag ttgcatgata aagaagacag tcataagtgc ggcgacgata
     3721 gtcatgcccc gcgcccaccg gaaggagctg actgggttga aggctctcaa gggcatcggt
     3781 cgagatcccg gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg ctcactgccc
     3841 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     3901 agaggcggtt tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg agacgggcaa
     3961 cagctgattg cccttcaccg cctggccctg agagagttgc agcaagcggt ccacgctggt
     4021 ttgccccagc aggcgaaaat cctgtttgat ggtggttaac ggcgggatat aacatgagct
     4081 gtcttcggta tcgtcgtatc ccactaccga gatatccgca ccaacgcgca gcccggactc
     4141 ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg
     4201 aacgatgccc tcattcagca tttgcatggt ttgttgaaaa ccggacatgg cactccagtc
     4261 gccttcccgt tccgctatcg gctgaatttg attgcgagtg agatatttat gccagccagc
     4321 cagacgcaga cgcgccgaga cagaacttaa tgggcccgct aacagcgcga tttgctggtg
     4381 acccaatgcg accagatgct ccacgcccag tcgcgtaccg tcttcatggg agaaaataat
     4441 actgttgatg ggtgtctggt cagagacatc aagaaataac gccggaacat tagtgcaggc
     4501 agcttccaca gcaatggcat cctggtcatc cagcggatag ttaatgatca gcccactgac
     4561 gcgttgcgcg agaagattgt gcaccgccgc tttacaggct tcgacgccgc ttcgttctac
     4621 catcgacacc accacgctgg cacccagttg atcggcgcga gatttaatcg ccgcgacaat
     4681 ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca acgactgttt
     4741 gcccgccagt tgttgtgcca cgcggttggg aatgtaattc agctccgcca tcgccgcttc
     4801 cactttttcc cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt
     4861 ctgataagag acaccggcat actctgcgac atcgtataac gttactggtt tcacattcac
     4921 caccctgaat tgactctctt ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca
     4981 ttcgatggtg tccgggatct cgacgctctc ccttatgcga ctcctgcatt aggaagcagc
     5041 ccagtagtag gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca tgcaaggaga
     5101 tggcgcccaa cagtcccccg gccacggggc ctgccaccat acccacgccg aaacaagcgc
     5161 tcatgagccc gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc
     5221 cagcaaccgc acctgtggcg ccggtgatgc cggccacgat gcgtccggcg tagaggatcg
     5281 agatctcgat cccgcgaaat taatacgact cactataggg gaattgtgag cggataacaa
     5341 ttcccctcta gaaataattt tgtttaactt taagaaggag atataccatg ggctctgata
     5401 aaattcatca tcatcatcat cacgaaaatc tttattttca aggtcatatg gctagcatgg
     5461 agatggaaaa ggagttcgag cagatcgaca agtccgggag ctgggcggcc atttaccagg
     5521 atatccgaca tgaagccagt gacttcccat gtagagtggc caagcttcct aagaacaaaa
     5581 accgaaatag gtacagagac gtcagtccct ttgaccatag tcggattaaa ctacatcaag
     5641 aagataatga ctatatcaac gctagtttga taaaaatgga agaagcccaa aggagttaca
     5701 ttcttaccca gggccctttg cctaacacat gcggtcactt ttgggagatg gtgtgggagc
     5761 agaaaagcag gggtgtcgtc atgctcaaca gagtgatgga gaaaggttcg ttaaaatgcg
     5821 cacaatactg gccacaaaaa gaagaaaaag agatgatctt tgaagacaca aatttgaaat
     5881 taacattgat ctctgaagat atcaagtcat attatacagt gcgacagcta gaattggaaa
     5941 accttacgac ccaagaaact cgagagatct tacatttcca ctataccaca tggcctgact
     6001 ttggagtccc tgaatcacca gcctcattct tgaactttct tttcaaagtc cgagagtcag
     6061 ggtcactcag cccggagcac gggcccgttg tggtgcactg cagtgcaggc atcggcaggt
     6121 ctggaacctt ctgtctggct gatacctgcc tcttgctgat ggacaagagg aaagaccctt
     6181 cttccgttga tatcaagaaa gtgctgttag aaatgagga
//
