LOCUS       sequence-92237-e        5793 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Bacterial Expression Vector .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5793)
  AUTHORS   Hopkins BD, Fine B, Steinbach N, Dendy M, Rapp Z, Shaw J, Pappas K, 
            Yu JS, Hodakoski C, Mense S, Klein J, Pegno S, Sulis ML, Goldstein 
            H, Amendolara B, Lei L, Maurer M, Bruce J, Canoll P, Hibshoosh H, 
            Parsons R
  TITLE     A secreted PTEN phosphatase that enters cells to alter signaling and
            survival.
  JOURNAL   Science. 2013 Jul 26;341(6144):399-402. doi: 
            10.1126/science.1234907. Epub 2013 Jun 6.
  PUBMED    23744781
REFERENCE   2  (bases 1 to 5793)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1126/science.1234907"; journalName: "Science"; date: 
            "2013-07-26- 26"; volume: "341"; issue: "6144"; pages: "399-402"
COMMENT     Sequence Label: JpExpress404 PTEN-LONG
FEATURES             Location/Qualifiers
     source          1..5793
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      86..674
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     575..594
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     828..845
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     943..960
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     944..960
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1417..1433
                     /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)."
     promoter        join(1458..1472,1473..1478,1479..1495,1496..1502)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    1478..1494
                     /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)."
     protein_bind    1508..1527
                     /label=lac operator (symmetric)
                     /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). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     CDS             3317..3358
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3368..3385
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3590..3609)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(3615..3634)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        3676..3767
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3768..3836,3837..4628)
                     /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(3986..4005)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             4711..5793
                     /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(4730..4749)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 ctcatgacca aaatccctta acgtgagtta cgcgcgcgtc gttccactga gcgtcagacc
       61 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
      121 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
      181 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
      241 tgtagccgta gttagcccac cacttcaaga actctgtagc accgcctaca tacctcgctc
      301 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
      361 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
      421 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
      481 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
      541 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
      601 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
      661 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
      721 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
      781 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
      841 gcgaggaagc ggaaggcgag agtagggaac tgccaggcat caaactaagc agaaggcccc
      901 tgacggatgg cctttttgcg tttctacaaa ctctttctgt gttgtaaaac gacggccagt
      961 cttaagctcg ggccccctgg gcggttctga taacgagtaa tcgttaatcc gcaaataacg
     1021 taaaaacccg cttcggcggg tttttttatg gggggagttt agggaaagag catttgtcag
     1081 aatatttaag ggcgcctgtc actttgcttg atatatgaga attatttaac cttataaatg
     1141 agaaaaaagc aacgcacttt aaataagata cgttgctttt tcgattgatg aacacctata
     1201 attaaactat tcatctatta tttatgattt tttgtatata caatatttct agtttgttaa
     1261 agagaattaa gaaaataaat ctcgaaaata ataaagggaa aatcagtttt tgatatcaaa
     1321 attatacatg tcaacgataa tacaaaatat aatacaaact ataagatgtt atcagtattt
     1381 attatgcatt tagaataaat tttgtgtcgc ccttaattgt gagcggataa caattacgag
     1441 cttcatgcac agtgaaatca tgaaaaattt atttgctttg tgagcggata acaattataa
     1501 tatgtggaat tgtgagcgct cacaattcca caacggtttc cctctagaaa taattttgtt
     1561 taacttttag gaggtaaaac atatgagcga gtccccagtc accatcagca gagcaggcaa
     1621 cgcaggtgaa ctggttagcc cgttgctgct gccgccgacg cgccgtcgtc gtcgccgtca
     1681 cattcagggt cctggtccgg tgctgaacct gccgtctgct gcagctgctc cgccggttgc
     1741 tcgtgcgcca gaggcagcgg gtggcggttc ccgttccgag gattatagct cctcgccgca
     1801 tagcgcagca gcggcagctc gtccgctggc ggcagaggaa aagcaggctc agtcgctgca
     1861 accgagctct agccgtcgtt ctagccacta cccagcggcc gttcagagcc aggcagccgc
     1921 agaacgtggc gcgagcgcga cggccaaaag ccgcgcaatt agcattctgc aaaagaaacc
     1981 gcgtcaccaa caactgttgc caagcttgag cagcttcttt ttcagccacc gtctgccgga
     2041 tatgaccgcg attatcaaag agatcgtgtc ccgtaacaag cgccgttatc aagaagatgg
     2101 ctttgacctg gacctgacct acatctaccc gaacattatc gcgatgggtt ttccagccga
     2161 gcgcctggag ggcgtctacc gtaataacat cgatgatgtg gttcgttttc tggacagcaa
     2221 acataagaac cattacaaga tctataatct gtgcgccgag cgtcattatg acaccgcgaa
     2281 attcaattgc cgtgttgcgc agtacccgtt cgaggaccac aatccgccgc aactggaact
     2341 gattaaaccg ttctgcgagg acctggatca gtggctgtcc gaggacgaca accacgtggc
     2401 cgcgattcac tgtaaggccg gcaaaggtcg taccggcgtc atgatctgcg cgtacctgct
     2461 gcaccgcggt aagtttctga aggcgcaaga agcgctggac ttttacggcg aggtgcgtac
     2521 ccgcgacaaa aagggtgtca ccatcccgag ccaacgccgc tatgtttact actacagcta
     2581 tttgctgaaa aaccacctgg attatcgtcc tgtcgccctg ttgtttcaca agatgatgtt
     2641 tgaaacgatt ccgatgttca gcggcggtac ctgtaatccg cagttcgtcg tttgtcagct
     2701 gaaggtgaag atctacagct ctaatagcgg tccgacccgt cgcgaagata agtttatgta
     2761 tttcgaattc ccgcagccgc tgccggtttg tggtgacatc aaggtggagt tcttccataa
     2821 acaaaacaag atgttgaaga aagataagat gttccatttt tgggtcaaca ctttctttat
     2881 tccgggtccg gaggagacta gcgagaaagt tgagaacggt agcctgtgcg atcaagaaat
     2941 cgatagcatt tgcagcatcg aacgtgcgga taatgacaaa gaatatctgg tcttgaccct
     3001 gacgaaaaac gacctggaca aagcgaacaa agataaagcg aatcgctatt tcagcccgaa
     3061 tttcaaagtt aagctgtact ttaccaaaac tgtggaggaa ccgagcaatc cggaggcgtc
     3121 tagctctacc agcgtgaccc cggacgtcag cgacaatgag ccggaccact atcgttacag
     3181 cgacacgacc gacagcgatc ctgaaaatga accgtttgac gaagatcaac atacgcagat
     3241 taccaaggtg aaaggcaaca gcgcggatat tcagcattct ggtggccgta gcagcttgga
     3301 aggccctcgc ttcgagggta agccgattcc gaatccgctg ctgggtctgg atagcacgcg
     3361 cacgggtcac caccaccatc accactaact cgagccccaa gggcgacacc ccataattag
     3421 cccgggcgaa aggcccagtc tttcgactga gcctttcgtt ttatttgatg cctggcagtt
     3481 ccctactctc gcatggggag tccccacact accatcggcg ctacggcgtt tcacttctga
     3541 gttcggcatg gggtcaggtg ggaccaccgc gctactgccg ccaggcaaac aaggggtgtt
     3601 atgagccata ttcaggtata aatgggctcg cgataatgtt cagaattggt taattggttg
     3661 taacactgac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     3721 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagaatatg agtattcaac
     3781 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     3841 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     3901 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     3961 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     4021 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     4081 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     4141 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     4201 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     4261 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcgatgg
     4321 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     4381 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     4441 ctggctggtt tattgctgat aaatccggag ccggtgagcg tggttctcgc ggtatcatcg
     4501 cagcgctggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     4561 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     4621 attggtaagc ggcgcgccat cgaatggcgc aaaacctttc gcggtatggc atgatagcgc
     4681 ccggaagaga gtcaattcag ggtggtgaat atgaaaccag taacgttata cgatgtcgca
     4741 gagtatgccg gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt
     4801 tctgcgaaaa cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta cattcccaac
     4861 cgcgtggcac aacaactggc gggcaaacag tcgttgctga ttggcgttgc cacctccagt
     4921 ctggccctgc acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg
     4981 ggtgccagcg tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg
     5041 gtgcacaatc ttctcgcgca acgcgtcagt gggctgatca ttaactatcc gctggatgac
     5101 caggatgcca ttgctgtgga agctgcctgc actaatgttc cggcgttatt tcttgatgtc
     5161 tctgaccaga cacccatcaa cagtattatt ttctcccatg aggacggtac gcgactgggc
     5221 gtggagcatc tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg cccattaagt
     5281 tctgtctcgg cgcgtctgcg tctggctggc tggcataaat atctcactcg caatcaaatt
     5341 cagccgatag cggaacggga aggcgactgg agtgccatgt ccggttttca acaaaccatg
     5401 caaatgctga atgagggcat cgttcccact gcgatgctgg ttgccaacga tcagatggcg
     5461 ctgggcgcaa tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga tatctcggta
     5521 gtgggatacg acgataccga agatagctca tgttatatcc cgccgttaac caccatcaaa
     5581 caggattttc gcctgctggg gcaaaccagc gtggaccgct tgctgcaact ctctcagggc
     5641 caggcggtga agggcaatca gctgttgcca gtctcactgg tgaaaagaaa aaccaccctg
     5701 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
     5761 cgacaggttt cccgactgga aagcgggcag tga
//
