LOCUS       sequence-163458-        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: 901 pLNCX myr HA Akt1
FEATURES             Location/Qualifiers
     source          1..8108
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     42..66
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             148..189
                     /codon_start=1
                     /product="N-myristoylation signal from Src kinase (Pellman 
                     et al., 1985; Kaplan et al., 1988)"
                     /label=myr
                     /translation="MGSSKSKPKDPSQR"
     CDS             208..234
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             235..1671
                     /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             1777..2370
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(2473..2495)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2581..2721
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2636..2655
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2736..2753)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2907..3495)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2987..3006)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3666..4457,4458..4526))
                     /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     4289..4308
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4527..4631)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4699..4717
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     LTR             5289..5464
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    5527..5726
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     primer_bind     6180..6202
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     6218..6234
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             6294..7088
                     /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(6348..6367)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6958..6977
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     enhancer        7570..7949
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(7950..8108,1..45)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     join(8104..8108,1..16)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctcgttt agtgaaccgt
       61 cagatcgcct ggagacgcca tccacgctgt tttgacctcc atagaagaca ccgggaccga
      121 tccagcctcc gcggccccaa gcttaccatg gggtcttcaa aatctaaacc aaaggacccc
      181 agccagcgcc ggcgcaggat ccgaggttac ccatacgacg tgccagatta cgccagcgac
      241 gtggctattg tgaaggaggg ttggctgcac aaacgagggg agtacatcaa gacctggcgg
      301 ccacgctact tcctcctcaa gaatgatggc accttcattg gctacaagga gcggccgcag
      361 gatgtggacc aacgtgaggc tcccctcaac aacttctctg tggcgcagtg ccagctgatg
      421 aagacggagc ggccccggcc caacaccttc atcatccgct gcctgcagtg gaccactgtc
      481 atcgaacgca ccttccatgt ggagactcct gaggagcggg aggagtggac aaccgccatc
      541 cagactgtgg ctgacggcct caagaagcag gaggaggagg agatggactt ccggtcgggc
      601 tcacccagtg acaactcagg ggctgaagag atggaggtgt ccctggccaa gcccaagcac
      661 cgcgtgacca tgaacgagtt tgagtacctg aagctgctgg gcaagggcac tttcggcaag
      721 gtgatcctgg tgaaggagaa ggccacaggc cgctactacg ccatgaagat cctcaagaag
      781 gaagtcatcg tagccaagga cgaagtggcc cacacactca ccgagaaccg cgtcctgcag
      841 aactccaggc accccttcct cacagccctg aagtactctt tccagaccca cgaccgcctc
      901 tgctttgtca tggagtacgc caacgggggc gagctgttct tccacctgtc ccgggagcgt
      961 gtgttctccg aggaccgggc ccgcttctat ggcgctgaga ttgtgtcagc cctggactac
     1021 ctgcactcgg agaagaacgt ggtgtaccgg gacctcaagc tggagaacct catgctggac
     1081 aaggacgggc acattaagat cacagacttc gggctgtgca aggaggggat caaggacggt
     1141 gccaccatga agaccttttg cggcacacct gagtacctgg cccccgaggt gctggaggac
     1201 aatgactacg gccgtgcagt ggactggtgg gggctgggcg tggtcatgta cgagatgatg
     1261 tgcggtcgcc tgcccttcta caaccaggac catgagaagc tttttgagct catcctcatg
     1321 gaggagatcc gcttcccgcg cacgcttggt cccgaggcca agtccttgct ttcagggctg
     1381 ctcaagaagg accccaagca gaggcttggc gggggctccg aggacgccaa ggagatcatg
     1441 cagcatcgct tctttgccgg tatcgtgtgg cagcacgtgt acgagaagaa gctcagccca
     1501 cccttcaagc cccaggtcac gtcggagact gacaccaggt attttgatga ggagttcacg
     1561 gcccagatga tcaccatcac accacctgac caagatgaca gcatggagtg tgtggacagc
     1621 gagcgcaggc cccacttccc ccagttctcc tactcggcca gcggcacggc ctgaggcggc
     1681 ggtggactgc ggaattcgat atcaagctta tcgataggcc taacacgagc catagataaa
     1741 ataaaagatt ttatttagtc tccagaaaaa ggggggaatg aaagacccca cctgtaggtt
     1801 tggcaagcta gcttaagtaa cgccattttg caaggcatgg aaaaatacat aactgagaat
     1861 agagaagttc agatcaaggt caggaacaga tggaacagct gaatatgggc caaacaggat
     1921 atctgtggta agcagttcct gccccggctc agggccaaga acagatggaa cagctgaata
     1981 tgggccaaac aggatatctg tggtaagcag ttcctgcccc ggctcagggc caagaacaga
     2041 tggtccccag atgcggtcca gccctcagca gtttctagag aaccatcaga tgtttccagg
     2101 gtgccccaag gacctgaaat gaccctgtgc cttatttgaa ctaaccaatc agttcgcttc
     2161 tcgcttctgt tcgcgcgctt ctgctccccg agctcaataa aagagcccac aacccctcac
     2221 tcggggcgcc agtcctccga ttgactgagt cgcccgggta cccgtgtatc caataaaccc
     2281 tcttgcagtt gcatccgact tgtggtctcg ctgttccttg ggagggtctc ctctgagtga
     2341 ttgactaccc gtcagcgggg gtctttcatt tgggggctcg tccgggatcg ggagacccct
     2401 gcccagggac caccgaccca ccaccgggag gtaagctggc tgcctcgcgc gtttcggtga
     2461 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
     2521 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
     2581 cgcagccatg acccagtcac gtagcgatag cggagtgtat actggcttaa ctatgcggca
     2641 tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta
     2701 aggagaaaat accgcatcag gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     2761 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     2821 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     2881 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     2941 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     3001 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     3061 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     3121 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     3181 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     3241 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     3301 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     3361 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     3421 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     3481 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     3541 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     3601 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     3661 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     3721 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     3781 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     3841 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     3901 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     3961 cgcaacgttg ttgccattgc tgcaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     4021 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     4081 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     4141 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     4201 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     4261 agttgctctt gcccggcgtc aacacgggat aataccgcgc cacatagcag aactttaaaa
     4321 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     4381 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     4441 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     4501 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     4561 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     4621 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
     4681 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtcttca agaattcata
     4741 ccagatcacc gaaaactgtc ctccaaatgt gtccccctca cactcccaaa ttcgcgggct
     4801 tctgcctctt agaccactct accctattcc ccacactcac cggagccaaa gccgcggccc
     4861 ttccgtttct ttgcttttga aagaccccac ccgtaggtgg caagctagct taagtaacgc
     4921 cactttgcaa ggcatggaaa aatacataac tgagaataga aaagttcaga tcaaggtcag
     4981 gaacaaagaa acagctgaat accaaacagg atatctgtgg taagcggttc ctgccccggc
     5041 tcagggccaa gaacagatga gacagctgag tgatgggcca aacaggatat ctgtggtaag
     5101 cagttcctgc cccggctcgg ggccaagaac agatggtccc cagatgcggt ccagccctca
     5161 gcagtttcta gtgaatcatc agatgtttcc agggtgcccc aaggacctga aaatgaccct
     5221 gtaccttatt tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc gcttccgctc
     5281 tccgagctca ataaaagagc ccacaacccc tcactcggcg cgccagtctt ccgatagact
     5341 gcgtcgcccg ggtacccgta ttcccaataa agcctcttgc tgtttgcatc cgaatcgtgg
     5401 tctcgctgtt ccttgggagg gtctcctctg agtgattgac tacccacgac gggggtcttt
     5461 catttggggg ctcgtccggg atttggagac ccctgcccag ggaccaccga cccaccaccg
     5521 ggaggtaagc tggccagcaa cttatctgtg tctgtccgat tgtctagtgt ctatgtttga
     5581 tgttatgcgc ctgcgtctgt actagttagc taactagctc tgtatctggc ggacccgtgg
     5641 tggaactgac gagttctgaa cacccggccg caaccctggg agacgtccca gggactttgg
     5701 gggccgtttt tgtggcccga cctgaggaag ggagtcgatg tggaatccga ccccgtcagg
     5761 atatgtggtt ctggtaggag acgagaacct aaaacagttc ccgcctccgt ctgaattttt
     5821 gctttcggtt tggaaccgaa gccgcgcgtc ttgtctgctg cagcgctgca gcatcgttct
     5881 gtgttgtctc tgtctgactg tgtttctgta tttgtctgaa aattagggcc agactgttac
     5941 cttctgctct gcagaatggc caacctttaa cgtcggatgg ccgcgagacg gcacctttaa
     6001 ccgagacctc atcacccagg ttaagatcaa ggtcttttca cctggcccgc atggacaccc
     6061 agaccaggtc ccctacatcg tgacctggga agccttggct tttgaccccc ctccctgggt
     6121 caagcccttt gtacacccta agcctccgcc tcctcttcct ccatccgccc cgtctctccc
     6181 ccttgaacct cctcgttcga ccccgcctcg atcctccctt tatccagccc tcactccttc
     6241 tctaggcgcc ggaattccga tctgatcaag agacaggatg aggatcgttt cgcatgattg
     6301 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     6361 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     6421 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg
     6481 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     6541 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     6601 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     6661 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     6721 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     6781 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg
     6841 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     6901 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     6961 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     7021 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     7081 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
     7141 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     7201 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     7261 cgggctcgat cccctcgcga gttggttcag ctgctgcctg aggctggacg acctcgcgga
     7321 gttctaccgg cagtgcaaat ccgtcggcat ccaggaaacc agcagcggct atccgcgcat
     7381 ccatgccccc gaactgcagg agtggggagg cacgatggcc gctttggtcg aggcggatcc
     7441 ggccattagc catattattc attggttata tagcataaat caatattggc tattggccat
     7501 tgcatacgtt gtatccatat cataatatgt acatttatat tggctcatgt ccaacattac
     7561 cgccatgttg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag
     7621 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
     7681 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
     7741 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
     7801 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
     7861 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     7921 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
     7981 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
     8041 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
     8101 tgacgcaa
//
