LOCUS       sequence-171231-        8551 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 8551)
  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 8551)
  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: 903 pLNCX HA Akt1
FEATURES             Location/Qualifiers
     source          1..8551
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    146..345
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             546..962
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     892..914
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     930..946
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1006..1800
                     /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(1060..1079)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1670..1689
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     enhancer        2282..2661
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2662..2865
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2816..2836
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2862..2886
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      2976..2985
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2988..3014
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3015..4451
                     /codon_start=1
                     /gene="AKT1"
                     /product="human serine/threonine protein kinase activated 
                     by growth factors"
                     /label=Akt1
                     /translation="SDVAIVKEGWLHKRGEYIKTWRPRYFLLKNDGTFIGYKERPQDVD
                     QREAPLNNFSVAQCQLMKTERPRPNTFIIRCLQWTTVIERTFHVETPEEREEWTTAIQT
                     VADGLKKQEEEEMDFRSGSPSDNSGAEEMEVSLAKPKHRVTMNEFEYLKLLGKGTFGKV
                     ILVKEKATGRYYAMKILKKEVIVAKDEVAHTLTENRVLQNSRHPFLTALKYSFQTHDRL
                     CFVMEYANGGELFFHLSRERVFSEDRARFYGAEIVSALDYLHSEKNVVYRDLKLENLML
                     DKDGHIKITDFGLCKEGIKDGATMKTFCGTPEYLAPEVLEDNDYGRAVDWWGLGVVMYE
                     MMCGRLPFYNQDHEKLFELILMEEIRFPRTLGPEAKSLLSGLLKKDPKQRLGGGSEDAK
                     EIMQHRFFAGIVWQHVYEKKLSPPFKPQVTSETDTRYFDEEFTAQMITITPPDQDDSME
                     CVDSERRPHFPQFSYSASGTA"
     LTR             4557..5150
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5253..5275)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    5361..5501
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     5416..5435
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5516..5533)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5687..6275)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5767..5786)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6446..7237,7238..7306))
                     /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     7069..7088
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7307..7411)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7479..7497
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     LTR             8069..8244
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    8307..8506
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
ORIGIN
        1 gtttgcatcc gaatcgtggt ctcgctgttc cttgggaggg tctcctctga gtgattgact
       61 acccacgacg ggggtctttc atttgggggc tcgtccggga tttggagacc cctgcccagg
      121 gaccaccgac ccaccaccgg gaggtaagct ggccagcaac ttatctgtgt ctgtccgatt
      181 gtctagtgtc tatgtttgat gttatgcgcc tgcgtctgta ctagttagct aactagctct
      241 gtatctggcg gacccgtggt ggaactgacg agttctgaac acccggccgc aaccctggga
      301 gacgtcccag ggactttggg ggccgttttt gtggcccgac ctgaggaagg gagtcgatgt
      361 ggaatccgac cccgtcagga tatgtggttc tggtaggaga cgagaaccta aaacagttcc
      421 cgcctccgtc tgaatttttg ctttcggttt ggaaccgaag ccgcgcgtct tgtctgctgc
      481 agcgctgcag catcgttctg tgttgtctct gtctgactgt gtttctgtat ttgtctgaaa
      541 attagggcca gactgttacc actcccttaa gtttgacctt aggtcactgg aaagatgtcg
      601 agcggatcgc tcacaaccag tcggtagatg tcaagaagag acgttgggtt accttctgct
      661 ctgcagaatg gccaaccttt aacgtcggat ggccgcgaga cggcaccttt aaccgagacc
      721 tcatcaccca ggttaagatc aaggtctttt cacctggccc gcatggacac ccagaccagg
      781 tcccctacat cgtgacctgg gaagccttgg cttttgaccc ccctccctgg gtcaagccct
      841 ttgtacaccc taagcctccg cctcctcttc ctccatccgc cccgtctctc ccccttgaac
      901 ctcctcgttc gaccccgcct cgatcctccc tttatccagc cctcactcct tctctaggcg
      961 ccggaattcc gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     1021 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     1081 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     1141 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     1201 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     1261 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     1321 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     1381 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     1441 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     1501 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     1561 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     1621 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     1681 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     1741 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     1801 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     1861 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     1921 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccgggctcg
     1981 atcccctcgc gagttggttc agctgctgcc tgaggctgga cgacctcgcg gagttctacc
     2041 ggcagtgcaa atccgtcggc atccaggaaa ccagcagcgg ctatccgcgc atccatgccc
     2101 ccgaactgca ggagtgggga ggcacgatgg ccgctttggt cgaggcggat ccggccatta
     2161 gccatattat tcattggtta tatagcataa atcaatattg gctattggcc attgcatacg
     2221 ttgtatccat atcataatat gtacatttat attggctcat gtccaacatt accgccatgt
     2281 tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc
     2341 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
     2401 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
     2461 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
     2521 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
     2581 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
     2641 ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag
     2701 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt
     2761 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa
     2821 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctcgttt agtgaaccgt
     2881 cagatcgcct ggagacgcca tccacgctgt tttgacctcc atagaagaca ccgggaccga
     2941 tccagcctcc gcggccccaa gctggaagat ccgccgccac catgggttac ccatacgacg
     3001 tgccagatta cgccagcgac gtggctattg tgaaggaggg ttggctgcac aaacgagggg
     3061 agtacatcaa gacctggcgg ccacgctact tcctcctcaa gaatgatggc accttcattg
     3121 gctacaagga gcggccgcag gatgtggacc aacgtgaggc tcccctcaac aacttctctg
     3181 tggcgcagtg ccagctgatg aagacggagc ggccccggcc caacaccttc atcatccgct
     3241 gcctgcagtg gaccactgtc atcgaacgca ccttccatgt ggagactcct gaggagcggg
     3301 aggagtggac aaccgccatc cagactgtgg ctgacggcct caagaagcag gaggaggagg
     3361 agatggactt ccggtcgggc tcacccagtg acaactcagg ggctgaagag atggaggtgt
     3421 ccctggccaa gcccaagcac cgcgtgacca tgaacgagtt tgagtacctg aagctgctgg
     3481 gcaagggcac tttcggcaag gtgatcctgg tgaaggagaa ggccacaggc cgctactacg
     3541 ccatgaagat cctcaagaag gaagtcatcg tagccaagga cgaagtggcc cacacactca
     3601 ccgagaaccg cgtcctgcag aactccaggc accccttcct cacagccctg aagtactctt
     3661 tccagaccca cgaccgcctc tgctttgtca tggagtacgc caacgggggc gagctgttct
     3721 tccacctgtc ccgggagcgt gtgttctccg aggaccgggc ccgcttctat ggcgctgaga
     3781 ttgtgtcagc cctggactac ctgcactcgg agaagaacgt ggtgtaccgg gacctcaagc
     3841 tggagaacct catgctggac aaggacgggc acattaagat cacagacttc gggctgtgca
     3901 aggaggggat caaggacggt gccaccatga agaccttttg cggcacacct gagtacctgg
     3961 cccccgaggt gctggaggac aatgactacg gccgtgcagt ggactggtgg gggctgggcg
     4021 tggtcatgta cgagatgatg tgcggtcgcc tgcccttcta caaccaggac catgagaagc
     4081 tttttgagct catcctcatg gaggagatcc gcttcccgcg cacgcttggt cccgaggcca
     4141 agtccttgct ttcagggctg ctcaagaagg accccaagca gaggcttggc gggggctccg
     4201 aggacgccaa ggagatcatg cagcatcgct tctttgccgg tatcgtgtgg cagcacgtgt
     4261 acgagaagaa gctcagccca cccttcaagc cccaggtcac gtcggagact gacaccaggt
     4321 attttgatga ggagttcacg gcccagatga tcaccatcac accacctgac caagatgaca
     4381 gcatggagtg tgtggacagc gagcgcaggc cccacttccc ccagttctcc tactcggcca
     4441 gcggcacggc ctgaggcggc ggtggactgc ggaattcgat atcaagctta tcgataggcc
     4501 taacacgagc catagataaa ataaaagatt ttatttagtc tccagaaaaa ggggggaatg
     4561 aaagacccca cctgtaggtt tggcaagcta gcttaagtaa cgccattttg caaggcatgg
     4621 aaaaatacat aactgagaat agagaagttc agatcaaggt caggaacaga tggaacagct
     4681 gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga
     4741 acagatggaa cagctgaata tgggccaaac aggatatctg tggtaagcag ttcctgcccc
     4801 ggctcagggc caagaacaga tggtccccag atgcggtcca gccctcagca gtttctagag
     4861 aaccatcaga tgtttccagg gtgccccaag gacctgaaat gaccctgtgc cttatttgaa
     4921 ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg agctcaataa
     4981 aagagcccac aacccctcac tcggggcgcc agtcctccga ttgactgagt cgcccgggta
     5041 cccgtgtatc caataaaccc tcttgcagtt gcatccgact tgtggtctcg ctgttccttg
     5101 ggagggtctc ctctgagtga ttgactaccc gtcagcgggg gtctttcatt tgggggctcg
     5161 tccgggatcg ggagacccct gcccagggac caccgaccca ccaccgggag gtaagctggc
     5221 tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg
     5281 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg
     5341 ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac gtagcgatag cggagtgtat
     5401 actggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg
     5461 aaataccgca cagatgcgta aggagaaaat accgcatcag gcgctcttcc gcttcctcgc
     5521 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     5581 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     5641 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     5701 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     5761 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     5821 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     5881 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     5941 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     6001 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     6061 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     6121 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     6181 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     6241 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     6301 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     6361 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     6421 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     6481 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     6541 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     6601 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     6661 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     6721 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tgcaggcatc gtggtgtcac
     6781 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     6841 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     6901 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     6961 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     7021 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aacacgggat aataccgcgc
     7081 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     7141 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     7201 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     7261 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     7321 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     7381 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     7441 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
     7501 ttcgtcttca agaattcata ccagatcacc gaaaactgtc ctccaaatgt gtccccctca
     7561 cactcccaaa ttcgcgggct tctgcctctt agaccactct accctattcc ccacactcac
     7621 cggagccaaa gccgcggccc ttccgtttct ttgcttttga aagaccccac ccgtaggtgg
     7681 caagctagct taagtaacgc cactttgcaa ggcatggaaa aatacataac tgagaataga
     7741 aaagttcaga tcaaggtcag gaacaaagaa acagctgaat accaaacagg atatctgtgg
     7801 taagcggttc ctgccccggc tcagggccaa gaacagatga gacagctgag tgatgggcca
     7861 aacaggatat ctgtggtaag cagttcctgc cccggctcgg ggccaagaac agatggtccc
     7921 cagatgcggt ccagccctca gcagtttcta gtgaatcatc agatgtttcc agggtgcccc
     7981 aaggacctga aaatgaccct gtaccttatt tgaactaacc aatcagttcg cttctcgctt
     8041 ctgttcgcgc gcttccgctc tccgagctca ataaaagagc ccacaacccc tcactcggcg
     8101 cgccagtctt ccgatagact gcgtcgcccg ggtacccgta ttcccaataa agcctcttgc
     8161 tgtttgcatc cgaatcgtgg tctcgctgtt ccttgggagg gtctcctctg agtgattgac
     8221 tacccacgac gggggtcttt catttggggg ctcgtccggg atttggagac ccctgcccag
     8281 ggaccaccga cccaccaccg ggaggtaagc tggccagcaa cttatctgtg tctgtccgat
     8341 tgtctagtgt ctatgtttga tgttatgcgc ctgcgtctgt actagttagc taactagctc
     8401 tgtatctggc ggacccgtgg tggaactgac gagttctgaa cacccggccg caaccctggg
     8461 agacgtccca gggactttgg gggccgtttt tgtggcccga cctgaggaag ggagtcgatg
     8521 tggaatccga ccccgtcagg atatgtggct g
//
