LOCUS       sequence-414032-        6206 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Bacterial expression of human PRL-1 C104D mutant without 
            farnesylation motif.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6206)
  AUTHORS   Kozlov G, Funato Y, Chen YS, Zhang Z, Illes K, Miki H, Gehring K
  TITLE     PRL3 pseudophosphatase activity is necessary and sufficient to 
            promote metastatic growth.
  JOURNAL   J Biol Chem. 2020 Aug 14;295(33):11682-11692. doi: 
            10.1074/jbc.RA120.014464. Epub 2020 Jun 22.
  PUBMED    32571875
REFERENCE   2  (bases 1 to 6206)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1074/jbc.RA120.014464"; journalName: "J Biol Chem"; date: 
            "2020-08-14- 14"; volume: "295"; issue: "33"; pages: "11682-11692"
COMMENT     Sequence Label: hPRL1_1-169_C104D
FEATURES             Location/Qualifiers
     source          1..6206
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(10..15,16..32,33..38)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     terminator      complement(213..260)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     256..274
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(838..855)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(865..882)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             901..923
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    938..962
                     /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(962..981)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(963..981)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1065..1084)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        1290..1367
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     complement(1387..1406)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             3761..3952
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(3946..3968)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4054..4193
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4109..4128
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4208..4225)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4379..4967)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4459..4478)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5138..5929,5930..5998))
                     /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     5761..5780
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5999..6103)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6171..6189
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ttctcatgtt tgacagctta tcatcgataa gctttaatgc ggtagtttat cacagttaaa
       61 ttgctaacgc agtcaggcac cgtgtatgaa atctaacaat gcgctcatcg tcatcctcgg
      121 caccgtcacc ctggatgctg taggcatagg cttggttatg ccggtactgc cgggcctctt
      181 gcgggatatc cggatatagt tcctcctttc agcaaaaaac ccctcaagac ccgtttagag
      241 gccccaaggg gttatgctag ttattgctca gcggtggcag cagccaactc agcttccttt
      301 cgggctttgt tagcagccgg atccttagtt gttacggtgg ccgttagaat ctttgaagcg
      361 cagacgcatt ttcggacggt atttttccag gtacagcagc tgtttagagt tgaacgcgcc
      421 acgacgtttc tgacggatga actgaaccgc atcttcgtat ttcatgccac cttcgatcag
      481 cgccagcgca accagaaccg gcgcacggcc cagacccgca acatcgtgaa ccgcgatgca
      541 gcagcccggt tcttcacgga atttgatttt aaccaggctc agccaatcat caacgatctg
      601 gttgctcggc ggcgcgccat catcgaacgg ccaatccaga acgtggatgc cttctttttc
      661 aaccagggtg gtatcgtagg tcgcttcgca aacacgaacg atggtggtaa caccgtattt
      721 tttcagttct tcgatgaatt tgttcagggt cgcgttggtc gggttgtggg tgatcaggaa
      781 tctcatgttt ttatatgtca cttcaaccgg cgctggtcta ttcattctag ccatatggct
      841 gccgcgcggc accaggccgc tgctgtgatg atgatgatga tggctgctgc ccatggtata
      901 tctccttctt aaagttaaac aaaattattt ctagagggga attgttatcc gctcacaatt
      961 cccctatagt gagtcgtatt aatttcgcgg gatcgagatc tcgatcctct acgccggacg
     1021 catcgtggcc ggcatcaccg gcgccacagg tgcggttgct ggcgcctata tcgccgacat
     1081 caccgatggg gaagatcggg ctcgccactt cgggctcatg agcgcttgtt tcggcgtggg
     1141 tatggtggca ggccccgtgg ccgggggact gttgggcgcc atctccttgc atgcaccatt
     1201 ccttgcggcg gcggtgctca acggcctcaa cctactactg ggctgcttcc taatgcagga
     1261 gtcgcataag ggagagcgtc gagatcccgg acaccatcga atggcgcaaa acctttcgcg
     1321 gtatggcatg atagcgcccg gaagagagtc aattcagggt ggtgaatgtg aaaccagtaa
     1381 cgttatacga tgtcgcagag tatgccggtg tctcttatca gaccgtttcc cgcgtggtga
     1441 accaggccag ccacgtttct gcgaaaacgc gggaaaaagt ggaagcggcg atggcggagc
     1501 tgaattacat tcccaaccgc gtggcacaac aactggcggg caaacagtcg ttgctgattg
     1561 gcgttgccac ctccagtctg gccctgcacg cgccgtcgca aattgtcgcg gcgattaaat
     1621 ctcgcgccga tcaactgggt gccagcgtgg tggtgtcgat ggtagaacga agcggcgtcg
     1681 aagcctgtaa agcggcggtg cacaatcttc tcgcgcaacg cgtcagtggg ctgatcatta
     1741 actatccgct ggatgaccag gatgccattg ctgtggaagc tgcctgcact aatgttccgg
     1801 cgttatttct tgatgtctct gaccagacac ccatcaacag tattattttc tcccatgaag
     1861 acggtacgcg actgggcgtg gagcatctgg tcgcattggg tcaccagcaa atcgcgctgt
     1921 tagcgggccc attaagttct gtctcggcgc gtctgcgtct ggctggctgg cataaatatc
     1981 tcactcgcaa tcaaattcag ccgatagcgg aacgggaagg cgactggagt gccatgtccg
     2041 gttttcaaca aaccatgcaa atgctgaatg agggcatcgt tcccactgcg atgctggttg
     2101 ccaacgatca gatggcgctg ggcgcaatgc gcgccattac cgagtccggg ctgcgcgttg
     2161 gtgcggatat ctcggtagtg ggatacgacg ataccgaaga cagctcatgt tatatcccgc
     2221 cgttaaccac catcaaacag gattttcgcc tgctggggca aaccagcgtg gaccgcttgc
     2281 tgcaactctc tcagggccag gcggtgaagg gcaatcagct gttgcccgtc tcactggtga
     2341 aaagaaaaac caccctggcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     2401 cattaatgca gctggcacga cagtttcccg actggaaagc gggcagtgag cgcaacgcaa
     2461 ttaatgtaag ttagctcact cattaggcac cgggatctcg accgatgccc ttgagagcct
     2521 tcaacccagt cagctccttc cggtgggcgc ggggcatgac tatcgtcgcc gcacttatga
     2581 ctgtcttctt tatcatgcaa ctcgtaggac aggtgccggc agcgctctgg gtcattttcg
     2641 gcgaggaccg ctttcgctgg agcgcgacga tgatcggcct gtcgcttgcg gtattcggaa
     2701 tcttgcacgc cctcgctcaa gccttcgtca ctggtcccgc caccaaacgt ttcggcgaga
     2761 agcaggccat tatcgccggc atggcggccg acgcgctggg ctacgtcttg ctggcgttcg
     2821 cgacgcgagg ctggatggcc ttccccatta tgattcttct cgcttccggc ggcatcggga
     2881 tgcccgcgtt gcaggccatg ctgtccaggc aggtagatga cgaccatcag ggacagcttc
     2941 aaggatcgct cgcggctctt accagcctaa cttcgatcat tggaccgctg atcgtcacgg
     3001 cgatttatgc cgcctcggcg agcacatgga acgggttggc atggattgta ggcgccgccc
     3061 tataccttgt ctgcctcccc gcgttgcgtc gcggtgcatg gagccgggcc acctcgacct
     3121 gaatggaagc cggcggcacc tcgctaacgg attcaccact ccaagaattg gagccaatca
     3181 attcttgcgg agaactgtga atgcgcaaac caacccttgg cagaacatat ccatcgcgtc
     3241 cgccatctcc agcagccgca cgcggcgcat ctcgggcagc gttgggtcct ggccacgggt
     3301 gcgcatgatc gtgctcctgt cgttgaggac ccggctaggc tggcggggtt gccttactgg
     3361 ttagcagaat gaatcaccga tacgcgagcg aacgtgaagc gactgctgct gcaaaacgtc
     3421 tgcgacctga gcaacaacat gaatggtctt cggtttccgt gtttcgtaaa gtctggaaac
     3481 gcggaagtca gcgccctgca ccattatgtt ccggatctgc atcgcaggat gctgctggct
     3541 accctgtgga acacctacat ctgtattaac gaagcgctgg cattgaccct gagtgatttt
     3601 tctctggtcc cgccgcatcc ataccgccag ttgtttaccc tcacaacgtt ccagtaaccg
     3661 ggcatgttca tcatcagtaa cccgtatcgt gagcatcctc tctcgtttca tcggtatcat
     3721 tacccccatg aacagaaatc ccccttacac ggaggcatca gtgaccaaac aggaaaaaac
     3781 cgcccttaac atggcccgct ttatcagaag ccagacatta acgcttctgg agaaactcaa
     3841 cgagctggac gcggatgaac aggcagacat ctgtgaatcg cttcacgacc acgctgatga
     3901 gctttaccgc agctgcctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     3961 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     4021 gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga
     4081 tagcggagtg tatactggct taactatgcg gcatcagagc agattgtact gagagtgcac
     4141 cattgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgctctt
     4201 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     4261 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     4321 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     4381 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     4441 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     4501 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     4561 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     4621 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     4681 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     4741 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     4801 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     4861 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     4921 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     4981 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     5041 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     5101 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     5161 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     5221 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     5281 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     5341 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     5401 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctgcaggca
     5461 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     5521 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     5581 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     5641 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     5701 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaacacggg
     5761 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     5821 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     5881 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     5941 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     6001 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     6061 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     6121 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     6181 tcacgaggcc ctttcgtctt caagaa
//
