LOCUS       sequence-132750-        5367 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 5367)
  AUTHORS   Ramaswamy S, Nakamura N, Vazquez F, Batt DB, Perera S, Roberts TM, 
            Sellers WR
  TITLE     Regulation of G1 progression by the PTEN tumor suppressor protein is
            linked to inhibition of the phosphatidylinositol 3-kinase/Akt 
            pathway.
  JOURNAL   Proc Natl Acad Sci U S A. 1999 Mar 2. 96(5):2110-5.
  PUBMED    10051603
REFERENCE   2  (bases 1 to 5367)
  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: "Proc Natl 
            Acad Sci U S A. 1999 Mar 2. 96(5):2110-5."
COMMENT     Sequence Label: 811 pSG5L HA PTEN C124S
FEATURES             Location/Qualifiers
     source          1..5367
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..27
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             7..33
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YDVPDYASL"
     polyA_signal    1301..1422
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1338..1357)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1392..1411
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1456..1473)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1627..2215)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1707..1726)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2386..3177,3178..3246))
                     /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     3009..3028
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3247..3351)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3419..3437
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3475..3497)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3597..3616
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(3683..4138)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3820..3841)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4032..4051
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4264..4286
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4278..4295
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4279..4295
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4332..4352)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4440..4650
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        4462..4658
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4509..4644
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          4689..5261
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     5269..5288
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     5316..5335
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5316..5334
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 tacccctacg acgtgcccga ctacgcctcc ctcggatcca tgacagccat catcaaagag
       61 atcgttagca gaaacaaaag gagatatcaa gaggatggat tcgacttaga cttgacctat
      121 atttatccaa acattattgc tatgggattt cctgcagaaa gacttgaagg cgtatacagg
      181 aacaatattg atgatgtagt aaggtttttg gattcaaagc ataaaaacca ttacaagata
      241 tacaatcttt gtgctgaaag acattatgac accgccaaat ttaattgcag agttgcacaa
      301 tatccttttg aagaccataa cccaccacag ctagaactta tcaaaccctt ttgtgaagat
      361 cttgaccaat ggctaagtga agatgacaat catgttgcag caattcactc caaggctgga
      421 aagggacgaa ctggtgtaat gatatgtgca tatttattac atcggggcaa atttttaaag
      481 gcacaagagg ccctagattt ctatggggaa gtaaggacca gagacaaaaa gggagtaact
      541 attcccagtc agaggcgcta tgtgtattat tatagctacc tgttaaagaa tcatctggat
      601 tatagaccag tggcactgtt gtttcacaag atgatgtttg aaactattcc aatgttcagt
      661 ggcggaactt gcaatcctca gtttgtggtc tgccagctaa aggtgaagat atattcctcc
      721 aattcaggac ccacacgacg ggaagacaag ttcatgtact ttgagttccc tcagccgtta
      781 cctgtgtgtg gtgatatcaa agtagagttc ttccacaaac agaacaagat gctaaaaaag
      841 gacaaaatgt ttcacttttg ggtaaataca ttcttcatac caggaccaga ggaaacctca
      901 gaaaaagtag aaaatggaag tctatgtgat caagaaatcg atagcatttg cagtatagag
      961 cgtgcagata atgacaagga atatctagta cttactttaa caaaaaatga tcttgacaaa
     1021 gcaaataaag acaaagccaa ccgatacttt tctccaaatt ttaaggtgaa gctgtacttc
     1081 acaaaaacag tagaggagcc gtcaaatcca gaggctagca gttcaacttc tgtaacacca
     1141 gatgttagtg acaatgaacc tgatcattat agatattctg acaccactga ctctgatcca
     1201 gagaatgaac cttttgatga agatcagcat acacaaatta caaaagtctg agaattcccc
     1261 gatatcgcct cgagtctaga tccagatctt attaaagcag aacttgttta ttgcagctta
     1321 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     1381 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggtcgactct
     1441 agactcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     1501 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     1561 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     1621 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     1681 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     1741 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     1801 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     1861 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     1921 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     1981 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     2041 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     2101 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     2161 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     2221 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     2281 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     2341 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     2401 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     2461 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     2521 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     2581 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     2641 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     2701 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     2761 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     2821 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     2881 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     2941 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     3001 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     3061 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     3121 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     3181 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     3241 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     3301 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     3361 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     3421 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg
     3481 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     3541 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc
     3601 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt
     3661 aaggagaaaa taccgcatca ggaaattgta agcgttaata ttttgttaaa attcgcgtta
     3721 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     3781 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     3841 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     3901 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     3961 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     4021 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     4081 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc
     4141 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
     4201 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
     4261 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt gtcgacttct gaggcggaaa
     4321 gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     4381 cagaagtatg caaagcatgc atctcaatta gtcagcaagg aaagtcccca ggctccccag
     4441 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     4501 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     4561 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     4621 agtgaagagg cttttttgga ggcctaggct tttgcaaaaa gctccggatc gatcctgaga
     4681 acttcagggt gagtttgggg acccttgatt gttctttctt tttcgctatt gtaaaattca
     4741 tgttatatgg agggggcaaa gttttcaggg tgttgtttag aatgggaaga tgtcccttgt
     4801 atcaccatgg accctcatga taattttgtt tctttcactt tctactctgt tgacaaccat
     4861 tgtctcctct tattttcttt tcattttctg taactttttc gttaaacttt agcttgcatt
     4921 tgtaacgaat ttttaaattc acttttgttt atttgtcaga ttgtaagtac tttctctaat
     4981 cacttttttt tcaaggcaat cagggtatat tatattgtac ttcagcacag ttttagagaa
     5041 caattgttat aattaaatga taaggtagaa tatttctgca tataaattct ggctggcgtg
     5101 gaaatattct tattggtaga aacaactaca tcctggtcat catcctgcct ttctctttat
     5161 ggttacaatg atatacactg tttgagatga ggataaaata ctctgagtcc aaaccgggcc
     5221 cctctgctaa ccatgttcat gccttcttct ttttcctaca gctcctgggc aacgtgctgg
     5281 ttattgtgct gtctcatcat tttggcaaag aattgtaata cgactcacta tagggcgaat
     5341 tgtcgacccg ggaagcttac catggcc
//
