LOCUS       40924_9956        6340 bp DNA     circular SYN 13-MAY-2021
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6340)
  AUTHORS   Rodriguez-Viciana P, Warne PH, Khwaja A, Marte BM, Pappin D, Das P, 
            Waterfield MD, Ridley A, Downward J
  TITLE     Role of phosphoinositide 3-OH kinase in cell transformation and 
            control of the actin cytoskeleton by Ras.
  JOURNAL   Cell. 1997 May 2;89(3):457-67.
  PUBMED    9150145
REFERENCE   2  (bases 1 to 6340)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 6340)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell."; 
            date: "1997-05-2"; volume: "89(3)"; pages: "457-67"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6340
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        143..173
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /label=lac operator
                     /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     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    complement(258..391)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(568..1359)
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     promoter        complement(1426..1755)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      complement(1769..2197)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     polyA_signal    complement(2243..2467)
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        2493..2511
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     CDS             complement(2887..3453)
                     /codon_start=1
                     /label=H-Ras (G12V)
                     /note="human oncoprotein generated by the G12V mutation in
                     the small GTPase H-Ras"
                     /translation="MTEYKLVVVGAVGVGKSALTIQLIQNHFVDEYDPTIEDSYRKQVV
                     IDGETCLLDILDTAGQEEYSAMRDQYMRTGEGFLCVFAINNTKSFEDIHQYREQIKRVK
                     DSDDVPMVLVGNKCDLAARTVESRQAQDLARSYGIPYIETSAKTRQGVEDAFYTLVREI
                     RQHKLRKLNPPDESGPGCMSCKCVLS"
     promoter        complement(3523..3541)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(3586..3789)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     enhancer        complement(3790..4169)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     4341..4360
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable
                     marker"
     promoter        4435..4539
                     /label=AmpR promoter
     CDS             4540..5397
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5571..6159
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6313..6330
                     /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 cccgatctcg
     2581 accttttttt ttttttttca agaccatcca ataatttact gtgatcccat ctgtgcccga
     2641 caagggccca cagaggcctg ggaggggagc taagggctgg ggttccggtg gcatttggga
     2701 tgttcaagac agtctgtgca cagcctccct gggagggtct gcagtcacct cggcccacgg
     2761 tcccggggtg actgggctcc agcagccctt ccttccttcc ttgcttccgt ccttccttcc
     2821 tcctccttcc gtctgcacct ccttcctgca tccggcacct ccatgtcctg agcttgtgct
     2881 gcgtcaggag agcacacact tgcagctcat gcagccgggg ccactctcat caggagggtt
     2941 cagcttccgc agcttgtgct gccggatctc acgcaccaac gtgtagaagg catcctccac
     3001 tccctgccgg gtcttggccg aggtctcgat gtaggggatg ccgtagcttc gggcgaggtc
     3061 ctgagcctgc cgagattcca cagtgcgtgc agccaggtca cacttgttcc ccaccagcac
     3121 catgggcacg tcatccgagt ccttcacccg tttgatctgc tccctgtact ggtggatgtc
     3181 ctcaaaagac ttggtgttgt tgatggcaaa cacacacagg aagccctccc cggtgcgcat
     3241 gtactggtcc cgcatggcgc tgtactcctc ctggccggcg gtatccagga tgtccaacag
     3301 gcacgtctcc ccatcaatga ccacctgctt ccggtaggaa tcctctatag tggggtcgta
     3361 ttcgtccaca aaatggttct ggatcagctg gatggtcagc gcactcttgc ccacaccgac
     3421 ggcgccaaca acaacaagct tgtattcggt catagatcgg gaattccagc acactggcgg
     3481 ccgttactag tggatccgag ctcggtacca agcttgggtc tccctatagt gagtcgtatt
     3541 aatttcgata agccagtaag cagtgggttc tctagttagc cagagagctc tgcttatata
     3601 gacctcccac cgtacacgcc taccgcccat ttgcgtcaat ggggcggagt tgttacgaca
     3661 ttttggaaag tcccgttgat tttggtgcca aaacaaactc ccattgacgt caatggggtg
     3721 gagacttgga aatccccgtg agtcaaaccg ctatccacgc ccattgatgt actgccaaaa
     3781 ccgcatcacc atggtaatag cgatgactaa tacgtagatg tactgccaag taggaaagtc
     3841 ccataaggtc atgtactggg cataatgcca ggcgggccat ttaccgtcat tgacgtcaat
     3901 agggggcgta cttggcatat gatacacttg atgtactgcc aagtgggcag tttaccgtaa
     3961 atactccacc cattgacgtc aatggaaagt ccctattggc gttactatgg gaacatacgt
     4021 cattattgac gtcaatgggc gggggtcgtt gggcggtcag ccaggcgggc catttaccgt
     4081 aagttatgta acgcggaact ccatatatgg gctatgaact aatgaccccg taattgatta
     4141 ctattaataa ctagtcaata atcaatgtca acgcgtatat ctggcccgta catcgcgaag
     4201 cagcgcaaaa cgcctaaccc taagcagatt cttcatgcaa ttgtcggtca agccttgcct
     4261 tgttgtagct taaattttgc tcgcgcacta ctcagcgacc tccaacacac aagcagggag
     4321 cagatactgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatagggg
     4381 atcgggagat ctcccgatcc gtcgacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     4441 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     4501 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     4561 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     4621 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     4681 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     4741 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     4801 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     4861 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     4921 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     4981 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     5041 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     5101 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     5161 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     5221 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     5281 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     5341 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     5401 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     5461 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     5521 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     5581 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     5641 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     5701 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     5761 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     5821 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     5881 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     5941 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     6001 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     6061 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     6121 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     6181 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     6241 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     6301 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
//
