LOCUS       sequence-139204-        5400 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 5400)
  TITLE     PTEN
REFERENCE   2  (bases 1 to 5400)
  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"
COMMENT     Sequence Label: 864 pSG5L HA NLS PTEN
FEATURES             Location/Qualifiers
     source          1..5400
                     /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"
     CDS             1962..1982
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    3256..3377
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3293..3312)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3347..3366
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3411..3428)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3582..4170)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3662..3681)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4341..5132,5133..5201))
                     /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     4964..4983
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5202..5306)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5374..5392
                     /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 tccgaagaag aagaggaagg
     1981 tcgaagacgg atccatgaca gccatcatca aagagatcgt tagcagaaac aaaaggagat
     2041 atcaagagga tggattcgac ttagacttga cctatattta tccaaacatt attgctatgg
     2101 gatttcctgc agaaagactt gaaggcgtat acaggaacaa tattgatgat gtagtaaggt
     2161 ttttggattc aaagcataaa aaccattaca agatatacaa tctttgtgct gaaagacatt
     2221 atgacaccgc caaatttaat tgcagagttg cacaatatcc ttttgaagac cataacccac
     2281 cacagctaga acttatcaaa cccttttgtg aagatcttga ccaatggcta agtgaagatg
     2341 acaatcatgt tgcagcaatt cactgtaaag ctggaaaggg acgaactggt gtaatgatat
     2401 gtgcatattt attacatcgg ggcaaatttt taaaggcaca agaggcccta gatttctatg
     2461 gggaagtaag gaccagagac aaaaagggag taactattcc cagtcagagg cgctatgtgt
     2521 attattatag ctacctgtta aagaatcatc tggattatag accagtggca ctgttgtttc
     2581 acaagatgat gtttgaaact attccaatgt tcagtggcgg aacttgcaat cctcagtttg
     2641 tggtctgcca gctaaaggtg aagatatatt cctccaattc aggacccaca cgacgggaag
     2701 acaagttcat gtactttgag ttccctcagc cgttacctgt gtgtggtgat atcaaagtag
     2761 agttcttcca caaacagaac aagatgctaa aaaaggacaa aatgtttcac ttttgggtaa
     2821 atacattctt cataccagga ccagaggaaa cctcagaaaa agtagaaaat ggaagtctat
     2881 gtgatcaaga aatcgatagc atttgcagta tagagcgtgc agataatgac aaggaatatc
     2941 tagtacttac tttaacaaaa aatgatcttg acaaagcaaa taaagacaaa gccaaccgat
     3001 acttttctcc aaattttaag gtgaagctgt acttcacaaa aacagtagag gagccgtcaa
     3061 atccagaggc tagcagttca acttctgtaa caccagatgt tagtgacaat gaacctgatc
     3121 attatagata ttctgacacc actgactctg atccagagaa tgaacctttt gatgaagatc
     3181 agcatacaca aattacaaaa gtctgagaat tccccgatat cgcctcgagt ctagatccag
     3241 atcttattaa agcagaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     3301 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     3361 ctcatcaatg tatcttatca tgtctggtcg actctagact cttccgcttc ctcgctcact
     3421 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     3481 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     3541 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     3601 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     3661 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     3721 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     3781 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     3841 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     3901 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     3961 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     4021 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     4081 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     4141 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     4201 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     4261 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     4321 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     4381 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     4441 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     4501 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     4561 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     4621 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     4681 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     4741 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     4801 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     4861 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     4921 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     4981 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     5041 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     5101 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     5161 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     5221 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     5281 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     5341 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
//
