LOCUS       sequence-182778-        8108 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 8108)
  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 8108)
  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: 902 pLNCX myr HA Akt1 K179M
FEATURES             Location/Qualifiers
     source          1..8108
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             414..589
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    652..851
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     primer_bind     1305..1327
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1343..1359
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1419..2213
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1473..1492)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2083..2102
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     enhancer        2695..3074
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3075..3278
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3229..3249
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     3275..3299
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             3381..3422
                     /codon_start=1
                     /product="N-myristoylation signal from Src kinase (Pellman 
                     et al., 1985; Kaplan et al., 1988)"
                     /label=myr
                     /translation="MGSSKSKPKDPSQR"
     CDS             3441..3467
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3468..4904
                     /codon_start=1
                     /gene="AKT1"
                     /product="human serine/threonine protein kinase activated 
                     by growth factors"
                     /label=Akt1
                     /translation="SDVAIVKEGWLHKRGEYIKTWRPRYFLLKNDGTFIGYKERPQDVD
                     QREAPLNNFSVAQCQLMKTERPRPNTFIIRCLQWTTVIERTFHVETPEEREEWTTAIQT
                     VADGLKKQEEEEMDFRSGSPSDNSGAEEMEVSLAKPKHRVTMNEFEYLKLLGKGTFGKV
                     ILVKEKATGRYYAMMILKKEVIVAKDEVAHTLTENRVLQNSRHPFLTALKYSFQTHDRL
                     CFVMEYANGGELFFHLSRERVFSEDRARFYGAEIVSALDYLHSEKNVVYRDLKLENLML
                     DKDGHIKITDFGLCKEGIKDGATMKTFCGTPEYLAPEVLEDNDYGRAVDWWGLGVVMYE
                     MMCGRLPFYNQDHEKLFELILMEEIRFPRTLGPEAKSLLSGLLKKDPKQRLGGGSEDAK
                     EIMQHRFFAGIVWQHVYEKKLSPPFKPQVTSETDTRYFDEEFTAQMITITPPDQDDSME
                     CVDSERRPHFPQFSYSASGTA"
     LTR             5010..5603
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5706..5728)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    5814..5954
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     5869..5888
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5969..5986)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6140..6728)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6220..6239)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6899..7690,7691..7759))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7522..7541
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7760..7864)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7932..7950
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tttgaaagac cccacccgta ggtggcaagc tagcttaagt aacgccactt tgcaaggcat
       61 ggaaaaatac ataactgaga atagaaaagt tcagatcaag gtcaggaaca aagaaacagc
      121 tgaataccaa acaggatatc tgtggtaagc ggttcctgcc ccggctcagg gccaagaaca
      181 gatgagacag ctgagtgatg ggccaaacag gatatctgtg gtaagcagtt cctgccccgg
      241 ctcggggcca agaacagatg gtccccagat gcggtccagc cctcagcagt ttctagtgaa
      301 tcatcagatg tttccagggt gccccaagga cctgaaaatg accctgtacc ttatttgaac
      361 taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc cgctctccga gctcaataaa
      421 agagcccaca acccctcact cggcgcgcca gtcttccgat agactgcgtc gcccgggtac
      481 ccgtattccc aataaagcct cttgctgttt gcatccgaat cgtggtctcg ctgttccttg
      541 ggagggtctc ctctgagtga ttgactaccc acgacggggg tctttcattt gggggctcgt
      601 ccgggatttg gagacccctg cccagggacc accgacccac caccgggagg taagctggcc
      661 agcaacttat ctgtgtctgt ccgattgtct agtgtctatg tttgatgtta tgcgcctgcg
      721 tctgtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa ctgacgagtt
      781 ctgaacaccc ggccgcaacc ctgggagacg tcccagggac tttgggggcc gtttttgtgg
      841 cccgacctga ggaagggagt cgatgtggaa tccgaccccg tcaggatatg tggttctggt
      901 aggagacgag aacctaaaac agttcccgcc tccgtctgaa tttttgcttt cggtttggaa
      961 ccgaagccgc gcgtcttgtc tgctgcagcg ctgcagcatc gttctgtgtt gtctctgtct
     1021 gactgtgttt ctgtatttgt ctgaaaatta gggccagact gttaccttct gctctgcaga
     1081 atggccaacc tttaacgtcg gatggccgcg agacggcacc tttaaccgag acctcatcac
     1141 ccaggttaag atcaaggtct tttcacctgg cccgcatgga cacccagacc aggtccccta
     1201 catcgtgacc tgggaagcct tggcttttga cccccctccc tgggtcaagc cctttgtaca
     1261 ccctaagcct ccgcctcctc ttcctccatc cgccccgtct ctcccccttg aacctcctcg
     1321 ttcgaccccg cctcgatcct ccctttatcc agccctcact ccttctctag gcgccggaat
     1381 tccgatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     1441 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     1501 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     1561 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     1621 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     1681 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     1741 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     1801 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     1861 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     1921 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     1981 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     2041 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     2101 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     2161 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     2221 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     2281 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     2341 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccgggc tcgatcccct
     2401 cgcgagttgg ttcagctgct gcctgaggct ggacgacctc gcggagttct accggcagtg
     2461 caaatccgtc ggcatccagg aaaccagcag cggctatccg cgcatccatg cccccgaact
     2521 gcaggagtgg ggaggcacga tggccgcttt ggtcgaggcg gatccggcca ttagccatat
     2581 tattcattgg ttatatagca taaatcaata ttggctattg gccattgcat acgttgtatc
     2641 catatcataa tatgtacatt tatattggct catgtccaac attaccgcca tgttgacatt
     2701 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
     2761 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
     2821 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
     2881 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
     2941 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
     3001 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
     3061 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
     3121 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
     3181 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
     3241 gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac cgtcagatcg
     3301 cctggagacg ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc
     3361 tccgcggccc caagcttacc atggggtctt caaaatctaa accaaaggac cccagccagc
     3421 gccggcgcag gatccgaggt tacccatacg acgtgccaga ttacgccagc gacgtggcta
     3481 ttgtgaagga gggttggctg cacaaacgag gggagtacat caagacctgg cggccacgct
     3541 acttcctcct caagaatgat ggcaccttca ttggctacaa ggagcggccg caggatgtgg
     3601 accaacgtga ggctcccctc aacaacttct ctgtggcgca gtgccagctg atgaagacgg
     3661 agcggccccg gcccaacacc ttcatcatcc gctgcctgca gtggaccact gtcatcgaac
     3721 gcaccttcca tgtggagact cctgaggagc gggaggagtg gacaaccgcc atccagactg
     3781 tggctgacgg cctcaagaag caggaggagg aggagatgga cttccggtcg ggctcaccca
     3841 gtgacaactc aggggctgaa gagatggagg tgtccctggc caagcccaag caccgcgtga
     3901 ccatgaacga gtttgagtac ctgaagctgc tgggcaaggg cactttcggc aaggtgatcc
     3961 tggtgaagga gaaggccaca ggccgctact acgccatgat gatcctcaag aaggaagtca
     4021 tcgtagccaa ggacgaagtg gcccacacac tcaccgagaa ccgcgtcctg cagaactcca
     4081 ggcacccctt cctcacagcc ctgaagtact ctttccagac ccacgaccgc ctctgctttg
     4141 tcatggagta cgccaacggg ggcgagctgt tcttccacct gtcccgggag cgtgtgttct
     4201 ccgaggaccg ggcccgcttc tatggcgctg agattgtgtc agccctggac tacctgcact
     4261 cggagaagaa cgtggtgtac cgggacctca agctggagaa cctcatgctg gacaaggacg
     4321 ggcacattaa gatcacagac ttcgggctgt gcaaggaggg gatcaaggac ggtgccacca
     4381 tgaagacctt ttgcggcaca cctgagtacc tggcccccga ggtgctggag gacaatgact
     4441 acggccgtgc agtggactgg tgggggctgg gcgtggtcat gtacgagatg atgtgcggtc
     4501 gcctgccctt ctacaaccag gaccatgaga agctttttga gctcatcctc atggaggaga
     4561 tccgcttccc gcgcacgctt ggtcccgagg ccaagtcctt gctttcaggg ctgctcaaga
     4621 aggaccccaa gcagaggctt ggcgggggct ccgaggacgc caaggagatc atgcagcatc
     4681 gcttctttgc cggtatcgtg tggcagcacg tgtacgagaa gaagctcagc ccacccttca
     4741 agccccaggt cacgtcggag actgacacca ggtattttga tgaggagttc acggcccaga
     4801 tgatcaccat cacaccacct gaccaagatg acagcatgga gtgtgtggac agcgagcgca
     4861 ggccccactt cccccagttc tcctactcgg ccagcggcac ggcctgaggc ggcggtggac
     4921 tgcggaattc gatatcaagc ttatcgatag gcctaacacg agccatagat aaaataaaag
     4981 attttattta gtctccagaa aaagggggga atgaaagacc ccacctgtag gtttggcaag
     5041 ctagcttaag taacgccatt ttgcaaggca tggaaaaata cataactgag aatagagaag
     5101 ttcagatcaa ggtcaggaac agatggaaca gctgaatatg ggccaaacag gatatctgtg
     5161 gtaagcagtt cctgccccgg ctcagggcca agaacagatg gaacagctga atatgggcca
     5221 aacaggatat ctgtggtaag cagttcctgc cccggctcag ggccaagaac agatggtccc
     5281 cagatgcggt ccagccctca gcagtttcta gagaaccatc agatgtttcc agggtgcccc
     5341 aaggacctga aatgaccctg tgccttattt gaactaacca atcagttcgc ttctcgcttc
     5401 tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc cacaacccct cactcggggc
     5461 gccagtcctc cgattgactg agtcgcccgg gtacccgtgt atccaataaa ccctcttgca
     5521 gttgcatccg acttgtggtc tcgctgttcc ttgggagggt ctcctctgag tgattgacta
     5581 cccgtcagcg ggggtctttc atttgggggc tcgtccggga tcgggagacc cctgcccagg
     5641 gaccaccgac ccaccaccgg gaggtaagct ggctgcctcg cgcgtttcgg tgatgacggt
     5701 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc
     5761 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc
     5821 atgacccagt cacgtagcga tagcggagtg tatactggct taactatgcg gcatcagagc
     5881 agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa
     5941 aataccgcat caggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     6001 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     6061 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     6121 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     6181 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     6241 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     6301 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     6361 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     6421 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     6481 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     6541 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     6601 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     6661 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     6721 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     6781 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     6841 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     6901 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     6961 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     7021 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     7081 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     7141 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     7201 ttgttgccat tgctgcaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     7261 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     7321 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     7381 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     7441 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     7501 cttgcccggc gtcaacacgg gataataccg cgccacatag cagaacttta aaagtgctca
     7561 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     7621 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     7681 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     7741 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     7801 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     7861 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
     7921 taacctataa aaataggcgt atcacgaggc cctttcgtct tcaagaattc ataccagatc
     7981 accgaaaact gtcctccaaa tgtgtccccc tcacactccc aaattcgcgg gcttctgcct
     8041 cttagaccac tctaccctat tccccacact caccggagcc aaagccgcgg cccttccgtt
     8101 tctttgct
//
