LOCUS       sequence-132808-        6897 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 6897)
  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 6897)
  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: 1014 pcDNA3 T7 Akt1 K179M T308A S473A
FEATURES             Location/Qualifiers
     source          1..6897
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    270..391
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(281..300)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     335..354
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(565..1359)
                     /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(676..695)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1286..1305
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(1426..1755)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1440..1575
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1494..1513)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     1740..1760
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(1769..2197)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1879..1900)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2091..2110
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    2243..2467
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2456..2473
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     promoter        2493..2511
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2493..2510
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             complement(2585..4024)
                     /codon_start=1
                     /gene="AKT1"
                     /product="human serine/threonine protein kinase activated 
                     by growth factors"
                     /label=Akt1
                     /translation="MSDVAIVKEGWLHKRGEYIKTWRPRYFLLKNDGTFIGYKERPQDV
                     DQREAPLNNFSVAQCQLMKTERPRPNTFIIRCLQWTTVIERTFHVETPEEREEWTTAIQ
                     TVADGLKKQEEEEMDFRSGSPSDNSGAEEMEVSLAKPKHRVTMNEFEYLKLLGKGTFGK
                     VILVKEKATGRYYAMMILKKEVIVAKDEVAHTLTENRVLQNSRHPFLTALKYSFQTHDR
                     LCFVMEYANGGELFFHLSRERVFSEDRARFYGAEIVSALDYLHSEKNVVYRDLKLENLM
                     LDKDGHIKITDFGLCKEGIKDGATMKAFCGTPEYLAPEVLEDNDYGRAVDWWGLGVVMY
                     EMMCGRLPFYNQDHEKLFELILMEEIRFPRTLGPEAKSLLSGLLKKDPKQRLGGGSEDA
                     KEIMQHRFFAGIVWQHVYEKKLSPPFKPQVTSETDTRYFDEEFTAQMITITPPDQDDSM
                     ECVDSERRPHFPQFAYSASGTA"
     regulatory      4057..4066
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(4079..4098)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4080..4098)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(4143..4346)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4172..4192)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4347..4726
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     4898..4917
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4992..5096
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5097..5165,5166..5957)
                     /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     complement(5315..5334)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6128..6716
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6617..6636
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6870..6887
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctct
      241 agctagaggt cgacggtata cagacatgat aagatacatt gatgagtttg gacaaaccac
      301 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
      361 tgtaaccatt ataagctgca ataaacaagt tggggtgggc gaagaactcc agcatgagat
      421 ccccgcgctg gaggatcatc cagccggcgt cccggaaaac gattccgaag cccaaccttt
      481 catagaaggc ggcggtggaa tcgaaatctc gtgatggcag gttgggcgtc gcttggtcgg
      541 tcatttcgaa ccccagagtc ccgctcagaa gaactcgtca agaaggcgat agaaggcgat
      601 gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg aagcggtcag cccattcgcc
      661 gccaagctct tcagcaatat cacgggtagc caacgctatg tcctgatagc ggtccgccac
      721 acccagccgg ccacagtcga tgaatccaga aaagcggcca ttttccacca tgatattcgg
      781 caagcaggca tcgccatggg tcacgacgag atcctcgccg tcgggcatgc gcgccttgag
      841 cctggcgaac agttcggctg gcgcgagccc ctgatgctct tcgtccagat catcctgatc
      901 gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg cgatgtttcg cttggtggtc
      961 gaatgggcag gtagccggat caagcgtatg cagccgccgc attgcatcag ccatgatgga
     1021 tactttctcg gcaggagcaa ggtgagatga caggagatcc tgccccggca cttcgcccaa
     1081 tagcagccag tcccttcccg cttcagtgac aacgtcgagc acagctgcgc aaggaacgcc
     1141 cgtcgtggcc agccacgata gccgcgctgc ctcgtcctgc agttcattca gggcaccgga
     1201 caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct gacagccgga acacggcggc
     1261 atcagagcag ccgattgtct gttgtgccca gtcatagccg aatagcctct ccacccaagc
     1321 ggccggagaa cctgcgtgca atccatcttg ttcaatcatg cgaaacgatc ctcatcctgt
     1381 ctcttgatca gatccgaaaa tggatataca agctcccggg agctttttgc aaaagcctag
     1441 gcctccaaaa aagcctcctc actacttctg gaatagctca gaggcagagg cggcctcggc
     1501 ctctgcataa ataaaaaaaa ttagtcagcc atggggcgga gaatgggcgg aactgggcgg
     1561 agttaggggc gggatgggcg gagttagggg cgggactatg gttgctgact aattgagatg
     1621 catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc tggttgctga
     1681 ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca
     1741 ccctaactga cacacattcc acagaattaa ttcgcgttaa atttttgtta aatcagctca
     1801 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     1861 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     1921 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     1981 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     2041 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     2101 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     2161 acacccgccg cgcttaatgc gccgctacag ggcgcgtggg gataccccct agagccccag
     2221 ctggttcttt ccgcctcaga agccatagag cccaccgcat ccccagcatg cctgctattg
     2281 tcttcccaat cctccccctt gctgtcctgc cccaccccac cccccagaat agaatgacac
     2341 ctactcagac aatgcgatgc aatttcctca ttttattagg aaaggacagt gggagtggca
     2401 ccttccaggg tcaaggaagg cacgggggag gggcaaacaa cagatggctg gcaactagaa
     2461 ggcacagtcg aggctgatca gcgagctcta gcatttaggt gacactatag aatagggccc
     2521 tctagatgca tgctcgagcg gccgccagtg tgatggatat ctgcagaatt ccaccgccgc
     2581 ctcaggccgt gccgctggcc gagtaggcga actgggggaa gtggggcctg cgctcgctgt
     2641 ccacacactc catgctgtca tcttggtcag gtggtgtgat ggtgatcatc tgggccgtga
     2701 actcctcatc aaaatacctg gtgtcagtct ccgacgtgac ctggggcttg aagggtgggc
     2761 tgagcttctt ctcgtacacg tgctgccaca cgataccggc aaagaagcga tgctgcatga
     2821 tctccttggc gtcctcggag cccccgccaa gcctctgctt ggggtccttc ttgagcagcc
     2881 ctgaaagcaa ggacttggcc tcgggaccaa gcgtgcgcgg gaagcggatc tcctccatga
     2941 ggatgagctc aaaaagcttc tcatggtcct ggttgtagaa gggcaggcga ccgcacatca
     3001 tctcgtacat gaccacgccc agcccccacc agtccactgc acggccgtag tcattgtcct
     3061 ccagcacctc gggggccagg tactcaggtg tgccgcaaaa ggccttcatg gtggcaccgt
     3121 ccttgatccc ctccttgcac agcccgaagt ctgtgatctt aatgtgcccg tccttgtcca
     3181 gcatgaggtt ctccagcttg aggtcccggt acaccacgtt cttctccgag tgcaggtagt
     3241 ccagggctga cacaatctca gcgccataga agcgggcccg gtcctcggag aacacacgct
     3301 cccgggacag gtggaagaac agctcgcccc cgttggcgta ctccatgaca aagcagaggc
     3361 ggtcgtgggt ctggaaagag tacttcaggg ctgtgaggaa ggggtgcctg gagttctgca
     3421 ggacgcggtt ctcggtgagt gtgtgggcca cttcgtcctt ggctacgatg acttccttct
     3481 tgaggatcat catggcgtag tagcggcctg tggccttctc cttcaccagg atcaccttgc
     3541 cgaaagtgcc cttgcccagc agcttcaggt actcaaactc gttcatggtc acgcggtgct
     3601 tgggcttggc cagggacacc tccatctctt cagcccctga gttgtcactg ggtgagcccg
     3661 accggaagtc catctcctcc tcctcctgct tcttgaggcc gtcagccaca gtctggatgg
     3721 cggttgtcca ctcctcccgc tcctcaggag tctccacatg gaaggtgcgt tcgatgacag
     3781 tggtccactg caggcagcgg atgatgaagg tgttgggccg gggccgctcc gtcttcatca
     3841 gctggcactg cgccacagag aagttgttga ggggagcctc acgttggtcc acatcctgcg
     3901 gccgctcctt gtagccaatg aaggtgccat cattcttgag gaggaagtag cgtggccgcc
     3961 aggtcttgat gtactcccct cgtttgtgca gccaaccctc cttcacaata gccacgtcgc
     4021 tcatggatcc catttgctgt ccaccagtca tgctagccat ggtggcaagc ttgggtctcc
     4081 ctatagtgag tcgtattaat ttcgataagc cagtaagcag tgggttctct agttagccag
     4141 agagctctgc ttatatagac ctcccaccgt acacgcctac cgcccatttg cgtcaatggg
     4201 gcggagttgt tacgacattt tggaaagtcc cgttgatttt ggtgccaaaa caaactccca
     4261 ttgacgtcaa tggggtggag acttggaaat ccccgtgagt caaaccgcta tccacgccca
     4321 ttgatgtact gccaaaaccg catcaccatg gtaatagcga tgactaatac gtagatgtac
     4381 tgccaagtag gaaagtccca taaggtcatg tactgggcat aatgccaggc gggccattta
     4441 ccgtcattga cgtcaatagg gggcgtactt ggcatatgat acacttgatg tactgccaag
     4501 tgggcagttt accgtaaata ctccacccat tgacgtcaat ggaaagtccc tattggcgtt
     4561 actatgggaa catacgtcat tattgacgtc aatgggcggg ggtcgttggg cggtcagcca
     4621 ggcgggccat ttaccgtaag ttatgtaacg cggaactcca tatatgggct atgaactaat
     4681 gaccccgtaa ttgattacta ttaataacta gtcaataatc aatgtcaacg cgtatatctg
     4741 gcccgtacat cgcgaagcag cgcaaaacgc ctaaccctaa gcagattctt catgcaattg
     4801 tcggtcaagc cttgccttgt tgtagcttaa attttgctcg cgcactactc agcgacctcc
     4861 aacacacaag cagggagcag atactggctt aactatgcgg catcagagca gattgtactg
     4921 agagtgcacc ataggggatc gggagatctc ccgatccgtc gacgtcaggt ggcacttttc
     4981 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     5041 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     5101 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     5161 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     5221 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     5281 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     5341 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     5401 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     5461 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     5521 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     5581 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     5641 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     5701 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     5761 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     5821 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     5881 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     5941 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     6001 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     6061 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     6121 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     6181 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     6241 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     6301 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     6361 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     6421 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     6481 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     6541 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     6601 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     6661 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     6721 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     6781 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     6841 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaag
//
