LOCUS       sequence-175309-        6876 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses halo tagged PTEN protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6876)
  AUTHORS   Fang X, Fu Y, Long MJ, Haegele JA, Ge EJ, Parvez S, Aye Y
  TITLE     Temporally controlled targeting of 4-hydroxynonenal to specific 
            proteins in living cells.
  JOURNAL   J Am Chem Soc. 2013 Oct 2;135(39):14496-9. doi: 10.1021/ja405400k. 
            Epub 2013 Sep 18.
  PUBMED    24015839
REFERENCE   2  (bases 1 to 6876)
  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: "10.1021/ja405400k";
            journalName: "J Am Chem Soc"; date: "2013-10-2- 2"; volume: "135"; 
            issue: "39"; pages: "14496-9"
COMMENT     Sequence Label: pMIR-His-Halo-Tev-PTEN
FEATURES             Location/Qualifiers
     source          1..6876
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             44..643
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(44..63)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     540..560
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    873..954
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(910..929)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        1042..1146
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1147..1215,1216..2007)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1365..1384)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2178..2766
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2667..2686
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2920..2937
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3054..3075
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3090..3095,3096..3113,3114..3120)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3128..3144
                     /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     3133..3155
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3152..3168
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3152..3168
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        3188..3206
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     3188..3205
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     3218..3237
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3218..3236
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     enhancer        3358..3661
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3662..3865
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3816..3836
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             3902..3919
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3929..3946
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             3950..4840
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
     CDS             4847..4867
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(6101..6118)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6101..6117)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6110..6132)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(6326..6345)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6445..6467
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        6507..6823
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6674..6809
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 ccctgaccca cgcccctgac ccctcacaag gagacgacct tccatgaccg agtacaagcc
       61 cacggtgcgc ctcgccaccc gcgacgacgt cccccgggcc gtacgcaccc tcgccgccgc
      121 gttcgccgac taccccgcca cgcgccacac cgtcgacccg gaccgccaca tcgagcgggt
      181 caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt
      241 cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc
      301 ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca
      361 gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc
      421 caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc
      481 cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg
      541 caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga
      601 aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgacgcccgc cccacgaccc
      661 gcagcgcccg accgaaagga gcgcacgacc ccatggctcc gaccgaagcc acccggggcg
      721 gccccgccga ccccgcaccc gcccccgagg cccaccgact ctagaggatc ataatcagcc
      781 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
      841 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
      901 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcaatctc
      961 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     1021 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     1081 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     1141 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     1201 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     1261 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     1321 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     1381 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     1441 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     1501 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     1561 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     1621 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     1681 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     1741 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     1801 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     1861 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     1921 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     1981 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     2041 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     2101 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     2161 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     2221 aaaaaaccac cgctaccagc ggtggttttt ttgccggatc aagagctacc aactcttttt
     2281 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     2341 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     2401 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     2461 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     2521 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     2581 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     2641 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     2701 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     2761 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     2821 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     2881 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     2941 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     3001 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg
     3061 agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg
     3121 tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacat gattacgaat
     3181 tgcaacgatt taggtgacac tatagaagag aaggaattaa tacgactcac tatagggaga
     3241 gagagagaat taccctcact aaagggagga gaagcatgaa ttcaaggtac cagatcttag
     3301 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
     3361 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
     3421 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
     3481 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
     3541 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
     3601 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
     3661 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
     3721 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
     3781 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
     3841 gtgggaggtc tatataagca gagctggttt agtgaaccgt ggatccacca tgggcagcag
     3901 ccatcatcat catcatcaca gcagcggcct ggtgcctcgt ggtagccata tggcagaaat
     3961 cggtactggc tttccattcg acccccatta tgtggaagtc ctgggcgagc gcatgcacta
     4021 cgtcgatgtt ggtccgcgcg atggcacccc tgtgctgttc ctgcacggta acccgacctc
     4081 ctcctacgtg tggcgcaaca tcatcccgca tgttgcaccg acccatcgct gcattgctcc
     4141 agacctgatc ggtatgggca aatccgacaa accagacctg ggttatttct tcgacgacca
     4201 cgtccgcttc atggatgcct tcatcgaagc cctgggtctg gaagaggtcg tcctggtcat
     4261 tcacgactgg ggctccgctc tgggtttcca ctgggccaag cgcaatccag agcgcgtcaa
     4321 aggtattgca tttatggagt tcatccgccc tatcccgacc tgggacgaat ggccagaatt
     4381 tgcccgcgag accttccagg ccttccgcac caccgacgtc ggccgcaagc tgatcatcga
     4441 tcagaacgtt tttatcgagg gtacgctgcc gatgggtgtc gtccgcccgc tgactgaagt
     4501 cgagatggac cattaccgcg agccgttcct gaatcctgtt gaccgcgagc cactgtggcg
     4561 cttcccaaac gagctgccaa tcgccggtga gccagcgaac atcgtcgcgc tggtcgaaga
     4621 atacatggac tggctgcacc agtcccctgt cccgaagctg ctgttctggg gcaccccagg
     4681 cgttctgatc ccaccggccg aagccgctcg cctggccaaa agcctgccta actgcaaggc
     4741 tgtggacatc ggcccgggtc tgaatctgct gcaagaagac aacccggacc tgatcggcag
     4801 cgagatcgcg cgctggctgt cgacgctcga gatttccggc tccggagaaa acttgtattt
     4861 ccagggctca gggatgacag ccatcatcaa agagatcgtt agcagaaaca aaaggagata
     4921 tcaagaggat ggattcgact tagacttgac ctatatttat ccaaacatta ttgctatggg
     4981 atttcctgca gaaagacttg aaggcgtata caggaacaat attgatgatg tagtaaggtt
     5041 tttggattca aagcataaaa accattacaa gatatacaat ctttgtgctg aaagacatta
     5101 tgacaccgcc aaatttaatt gcagagttgc acaatatcct tttgaagacc ataacccacc
     5161 acagctagaa cttatcaaac ccttttgtga agatcttgac caatggctaa gtgaagatga
     5221 caatcatgtt gcagcaattc actgtaaagc tggaaaggga cgaactggtg taatgatatg
     5281 tgcatattta ttacatcggg gcaaattttt aaaggcacaa gaggccctag atttctatgg
     5341 ggaagtaagg accagagaca aaaagggagt aactattccc agtcagaggc gctatgtgta
     5401 ttattatagc tacctgttaa agaatcatct ggattataga ccagtggcac tgttgtttca
     5461 caagatgatg tttgaaacta ttccaatgtt cagtggcgga acttgcaatc ctcagtttgt
     5521 ggtctgccag ctaaaggtga agatatattc ctccaattca ggacccacac gacgggaaga
     5581 caagttcatg tactttgagt tccctcagcc gttacctgtg tgtggtgata tcaaagtaga
     5641 gttcttccac aaacagaaca agatgctaaa aaaggacaaa atgtttcact tttgggtaaa
     5701 tacattcttc ataccaggac cagaggaaac ctcagaaaaa gtagaaaatg gaagtctatg
     5761 tgatcaagaa atcgatagca tttgcagtat agagcgtgca gataatgaca aggaatatct
     5821 agtacttact ttaacaaaaa atgatcttga caaagcaaat aaagacaaag ccaaccgata
     5881 cttttctcca aattttaagg tgaagctgta cttcacaaaa acagtagagg agccgtcaaa
     5941 tccagaggct agcagttcaa cttctgtaac accagatgtt agtgacaatg aacctgatca
     6001 ttatagatat tctgacacca ctgactctga tccagagaat gaaccttttg atgaagatca
     6061 gcatacacaa attacaaaag tctgaggccg ctagcttggc actggccgtc gttttacaac
     6121 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt
     6181 tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
     6241 gcctgaatgg cgaatggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     6301 cacaccgcat atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc
     6361 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     6421 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
     6481 caccgaaacg cgcgagacga aagattggtg tggaaagtcc ccaggctccc cagcaggcag
     6541 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     6601 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     6661 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     6721 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     6781 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctag cttgcatgcc
     6841 tgcaggtcgg ccgccacgac cggtgccgcc accatc
//
