LOCUS       sequence-162851-        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: 800 pSG5L HA PTEN wt
FEATURES             Location/Qualifiers
     source          1..5367
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(30..52)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     152..171
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(238..693)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(375..396)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     587..606
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     819..841
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     833..850
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     834..850
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(887..907)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        995..1205
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        1017..1213
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      1064..1199
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          1244..1816
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     1824..1843
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     1871..1890
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1871..1889
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1923..1949
                     /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="YPYDVPDYA"
     CDS             1923..1949
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1929..1955
                     /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    3223..3344
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3260..3279)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3314..3333
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3378..3395)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3549..4137)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3629..3648)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4308..5099,5100..5168))
                     /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     4931..4950
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5169..5273)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5341..5359
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
       61 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
      121 gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg
      181 caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggaaa
      241 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
      301 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
      361 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
      421 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
      481 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
      541 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
      601 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
      661 ccgccgcgct taatgcgccg ctacagggcg cgtccattcg ccattcaggc tgcgcaactg
      721 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg
      781 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac
      841 gacggccagt gaattgtcga cttctgaggc ggaaagaacc agctgtggaa tgtgtgtcag
      901 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
      961 aattagtcag caaggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     1021 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     1081 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     1141 cgaggccgcc tcggcctctg agctattcca gaagtagtga agaggctttt ttggaggcct
     1201 aggcttttgc aaaaagctcc ggatcgatcc tgagaacttc agggtgagtt tggggaccct
     1261 tgattgttct ttctttttcg ctattgtaaa attcatgtta tatggagggg gcaaagtttt
     1321 cagggtgttg tttagaatgg gaagatgtcc cttgtatcac catggaccct catgataatt
     1381 ttgtttcttt cactttctac tctgttgaca accattgtct cctcttattt tcttttcatt
     1441 ttctgtaact ttttcgttaa actttagctt gcatttgtaa cgaattttta aattcacttt
     1501 tgtttatttg tcagattgta agtactttct ctaatcactt ttttttcaag gcaatcaggg
     1561 tatattatat tgtacttcag cacagtttta gagaacaatt gttataatta aatgataagg
     1621 tagaatattt ctgcatataa attctggctg gcgtggaaat attcttattg gtagaaacaa
     1681 ctacatcctg gtcatcatcc tgcctttctc tttatggtta caatgatata cactgtttga
     1741 gatgaggata aaatactctg agtccaaacc gggcccctct gctaaccatg ttcatgcctt
     1801 cttctttttc ctacagctcc tgggcaacgt gctggttatt gtgctgtctc atcattttgg
     1861 caaagaattg taatacgact cactataggg cgaattgtcg acccgggaag cttaccatgg
     1921 cctaccccta cgacgtgccc gactacgcct ccctcggatc catgacagcc atcatcaaag
     1981 agatcgttag cagaaacaaa aggagatatc aagaggatgg attcgactta gacttgacct
     2041 atatttatcc aaacattatt gctatgggat ttcctgcaga aagacttgaa ggcgtataca
     2101 ggaacaatat tgatgatgta gtaaggtttt tggattcaaa gcataaaaac cattacaaga
     2161 tatacaatct ttgtgctgaa agacattatg acaccgccaa atttaattgc agagttgcac
     2221 aatatccttt tgaagaccat aacccaccac agctagaact tatcaaaccc ttttgtgaag
     2281 atcttgacca atggctaagt gaagatgaca atcatgttgc agcaattcac tgtaaagctg
     2341 gaaagggacg aactggtgta atgatatgtg catatttatt acatcggggc aaatttttaa
     2401 aggcacaaga ggccctagat ttctatgggg aagtaaggac cagagacaaa aagggagtaa
     2461 ctattcccag tcagaggcgc tatgtgtatt attatagcta cctgttaaag aatcatctgg
     2521 attatagacc agtggcactg ttgtttcaca agatgatgtt tgaaactatt ccaatgttca
     2581 gtggcggaac ttgcaatcct cagtttgtgg tctgccagct aaaggtgaag atatattcct
     2641 ccaattcagg acccacacga cgggaagaca agttcatgta ctttgagttc cctcagccgt
     2701 tacctgtgtg tggtgatatc aaagtagagt tcttccacaa acagaacaag atgctaaaaa
     2761 aggacaaaat gtttcacttt tgggtaaata cattcttcat accaggacca gaggaaacct
     2821 cagaaaaagt agaaaatgga agtctatgtg atcaagaaat cgatagcatt tgcagtatag
     2881 agcgtgcaga taatgacaag gaatatctag tacttacttt aacaaaaaat gatcttgaca
     2941 aagcaaataa agacaaagcc aaccgatact tttctccaaa ttttaaggtg aagctgtact
     3001 tcacaaaaac agtagaggag ccgtcaaatc cagaggctag cagttcaact tctgtaacac
     3061 cagatgttag tgacaatgaa cctgatcatt atagatattc tgacaccact gactctgatc
     3121 cagagaatga accttttgat gaagatcagc atacacaaat tacaaaagtc tgagaattcc
     3181 ccgatatcgc ctcgagtcta gatccagatc ttattaaagc agaacttgtt tattgcagct
     3241 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     3301 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggtcgact
     3361 ctagactctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     3421 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     3481 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     3541 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     3601 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     3661 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     3721 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     3781 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     3841 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     3901 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     3961 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     4021 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     4081 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     4141 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     4201 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     4261 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     4321 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     4381 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     4441 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     4501 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     4561 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     4621 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     4681 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     4741 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     4801 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     4861 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     4921 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     4981 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     5041 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     5101 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     5161 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     5221 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     5281 ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa
     5341 aataggcgta tcacgaggcc ctttcgt
//
