LOCUS       sequence-488721-        7979 bp    DNA     circular SYN 08-OCT-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7979)
  AUTHORS   Samuels Y, Wang Z, Bardelli A, Silliman N, Ptak J, Szabo S, Yan H, 
            Gazdar A, Powell SM, Riggins GJ, Willson JK, Markowitz S, Kinzler 
            KW, Vogelstein B, Velculescu VE
  TITLE     High frequency of mutations of the PIK3CA gene in human cancers.
  JOURNAL   Science. 2004 Apr 23. 304(5670):554.
  PUBMED    15016963
REFERENCE   2  (bases 1 to 7979)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 8, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Science. 
            2004 Apr 23. 304(5670):554."
COMMENT     Sequence Label: PIK3CA-WT (pFastBac)
FEATURES             Location/Qualifiers
     source          1..7979
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      2..456
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(89..108)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     298..319
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        482..586
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(587..655,656..1447)
                     /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(805..824)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1618..2206
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2107..2126
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2360..2377
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  2511..2735
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(2802..3335)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3524..3529,3530..3546,3547..3552))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        3904..3995
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     3920..3937
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             4062..4079
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             4101..4121
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    7469..7603
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7506..7525)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7560..7579
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  7632..7797
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
ORIGIN
        1 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
       61 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
      121 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
      181 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
      241 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt cttaatagtg
      301 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
      361 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
      421 acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt
      481 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
      541 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
      601 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
      661 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
      721 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
      781 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
      841 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
      901 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
      961 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     1021 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     1081 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     1141 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     1201 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     1261 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     1321 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     1381 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     1441 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     1501 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     1561 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     1621 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     1681 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     1741 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     1801 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     1861 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     1921 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     1981 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     2041 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     2101 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     2161 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     2221 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     2281 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     2341 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg
     2401 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atagaccagc cgcgtaacct
     2461 ggcaaaatcg gttacggttg agtaataaat ggatgccctg cgtaagcggg tgtgggcgga
     2521 caataaagtc ttaaactgaa caaaatagat ctaaactatg acaataaagt cttaaactag
     2581 acagaatagt tgtaaactga aatcagtcca gttatgctgt gaaaaagcat actggacttt
     2641 tgttatggct aaagcaaact cttcattttc tgaagtgcaa attgcccgtc gtattaaaga
     2701 ggggcgtggc caagggcatg gtaaagacta tattcgcggc gttgtgacaa tttaccgaac
     2761 aactccgcgg ccgggaagcc gatctcggct tgaacgaatt gttaggtggc ggtacttggg
     2821 tcgatatcaa agtgcatcac ttcttcccgt atgcccaact ttgtatagag agccactgcg
     2881 ggatcgtcac cgtaatctgc ttgcacgtag atcacataag caccaagcgc gttggcctca
     2941 tgcttgagga gattgatgag cgcggtggca atgccctgcc tccggtgctc gccggagact
     3001 gcgagatcat agatatagat ctcactacgc ggctgctcaa acttgggcag aacgtaagcc
     3061 gcgagagcgc caacaaccgc ttcttggtcg aaggcagcaa gcgcgatgaa tgtcttacta
     3121 cggagcaagt tcccgaggta atcggagtcc ggctgatgtt gggagtaggt ggctacgtct
     3181 ccgaactcac gaccgaaaag atcaagagca gcccgcatgg atttgacttg gtcagggccg
     3241 agcctacatg tgcgaatgat gcccatactt gagccaccta actttgtttt agggcgactg
     3301 ccctgctgcg taacatcgtt gctgctgcgt aacatcgttg ctgctccata acatcaaaca
     3361 tcgacccacg gcgtaacgcg cttgctgctt ggatgcccga ggcatagact gtacaaaaaa
     3421 acagtcataa caagccatga aaaccgccac tgcgccgtta ccaccgctgc gttcggtcaa
     3481 ggttctggac cagttgcgtg agcgcatacg ctacttgcat tacagtttac gaaccgaaca
     3541 ggcttatgtc aactgggttc gtgccttcat ccgtttccac ggtgtgcgtc acccggcaac
     3601 cttgggcagc agcgaagtcg aggcatttct gtcctggctg gcgaacgagc gcaaggtttc
     3661 ggtctccacg catcgtcagg cattggcggc cttgctgttc ttctacggca aggtgctgtg
     3721 cacggatctg ccctggcttc aggagatcgg aagacctcgg ccgtcgcggc gcttgccggt
     3781 ggtgctgacc ccggatgaag tggttcgcat cctcggtttt ctggaaggcg agcatcgttt
     3841 gttcgcccag gactctagct atagttctag tggttggcta cgtatactcc ggaatattaa
     3901 tagatcatgg agataattaa aatgataacc atctcgcaaa taaataagta ttttactgtt
     3961 ttcgtaacag ttttgtaata aaaaaaccta taaatattcc ggattattca taccgtccca
     4021 ccatcgggcg cggatctcgg tccgaaacca tgtcgtacta ccatcaccat caccatcacg
     4081 attacgatat cccaacgacc gaaaacctgt attttcaggg cgccatggga tccatgcctc
     4141 caagaccatc atcaggtgaa ctgtggggca tccacttgat gcccccaaga atcctagtag
     4201 aatgtttact accaaatgga atgatagtga ctttagaatg cctccgtgag gctacattaa
     4261 taaccataaa gcatgaacta tttaaagaag caagaaaata ccccctccat caacttcttc
     4321 aagatgaatc ttcttacatt ttcgtaagtg ttactcaaga agcagaaagg gaagaatttt
     4381 ttgatgaaac aagacgactt tgtgaccttc ggctttttca acccttttta aaagtaattg
     4441 aaccagtagg caaccgtgaa gaaaagatcc tcaatcgaga aattggtttt gctatcggca
     4501 tgccagtgtg tgaatttgat atggttaaag atccagaagt acaggacttc cgaagaaata
     4561 ttctgaacgt ttgtaaagaa gctgtggatc ttagggacct caattcacct catagtagag
     4621 caatgtatgt ctatcctcca aatgtagaat cttcaccaga attgccaaag cacatatata
     4681 ataaattaga taaagggcaa ataatagtgg tgatctgggt aatagtttct ccaaataatg
     4741 acaagcagaa gtatactctg aaaatcaacc atgactgtgt accagaacaa gtaattgctg
     4801 aagcaatcag gaaaaaaact cgaagtatgt tgctatcctc tgaacaacta aaactctgtg
     4861 ttttagaata tcagggcaag tatattttaa aagtgtgtgg atgtgatgaa tacttcctag
     4921 aaaaatatcc tctgagtcag tataagtata taagaagctg tataatgctt gggaggatgc
     4981 ccaatttgat gttgatggct aaagaaagcc tttattctca actgccaatg gactgtttta
     5041 caatgccatc ttattccaga cgcatttcca cagctacacc atatatgaat ggagaaacat
     5101 ctacaaaatc cctttgggtt ataaatagtg cactcagaat aaaaattctt tgtgcaacct
     5161 acgtgaatgt aaatattcga gacattgata agatctatgt tcgaacaggt atctaccatg
     5221 gaggagaacc cttatgtgac aatgtgaaca ctcaaagagt accttgttcc aatcccaggt
     5281 ggaatgaatg gctgaattat gatatataca ttcctgatct tcctcgtgct gctcgacttt
     5341 gcctttccat ttgctctgtt aaaggccgaa agggtgctaa agaggaacac tgtccattgg
     5401 catggggaaa tataaacttg tttgattaca cagacactct agtatctgga aaaatggctt
     5461 tgaatctttg gccagtacct catggattag aagatttgct gaaccctatt ggtgttactg
     5521 gatcaaatcc aaataaagaa actccatgct tagagttgga gtttgactgg ttcagcagtg
     5581 tggtaaagtt cccagatatg tcagtgattg aagagcatgc caattggtct gtatcccgag
     5641 aagcaggatt tagctattcc cacgcaggac tgagtaacag actagctaga gacaatgaat
     5701 taagggaaaa tgacaaagaa cagctcaaag caatttctac acgagatcct ctctctgaaa
     5761 tcactgagca ggagaaagat tttctatgga gtcacagaca ctattgtgta actatccccg
     5821 aaattctacc caaattgctt ctgtctgtta aatggaattc tagagatgaa gtagcccaga
     5881 tgtattgctt ggtaaaagat tggcctccaa tcaaacctga acaggctatg gaacttctgg
     5941 actgtaatta cccagatcct atggttcgag gttttgctgt tcggtgcttg gaaaaatatt
     6001 taacagatga caaactttct cagtatttaa ttcagctagt acaggtccta aaatatgaac
     6061 aatatttgga taacttgctt gtgagatttt tactgaagaa agcattgact aatcaaagga
     6121 ttgggcactt tttcttttgg catttaaaat ctgagatgca caataaaaca gttagccaga
     6181 ggtttggcct gcttttggag tcctattgtc gtgcatgtgg gatgtatttg aagcacctga
     6241 ataggcaagt cgaggcaatg gaaaagctca ttaacttaac tgacattctc aaacaggaga
     6301 agaaggatga aacacaaaag gtacagatga agtttttagt tgagcaaatg aggcgaccag
     6361 atttcatgga tgctctacag ggctttctgt ctcctctaaa ccctgctcat caactaggaa
     6421 acctcaggct tgaagagtgt cgaattatgt cctctgcaaa aaggccactg tggttgaatt
     6481 gggagaaccc agacatcatg tcagagttac tgtttcagaa caatgagatc atctttaaaa
     6541 atggggatga tttacggcaa gatatgctaa cacttcaaat tattcgtatt atggaaaata
     6601 tctggcaaaa tcaaggtctt gatcttcgaa tgttacctta tggttgtctg tcaatcggtg
     6661 actgtgtggg acttattgag gtggtgcgaa attctcacac tattatgcaa attcagtgca
     6721 aaggcggctt gaaaggtgca ctgcagttca acagccacac actacatcag tggctcaaag
     6781 acaagaacaa aggagaaata tatgatgcag ccattgacct gtttacacgt tcatgtgctg
     6841 gatactgtgt agctaccttc attttgggaa ttggagatcg tcacaatagt aacatcatgg
     6901 tgaaagacga tggacaactg tttcatatag attttggaca ctttttggat cacaagaaga
     6961 aaaaatttgg ttataaacga gaacgtgtgc catttgtttt gacacaggat ttcttaatag
     7021 tgattagtaa aggagcccaa gaatgcacaa agacaagaga atttgagagg tttcaggaga
     7081 tgtgttacaa ggcttatcta gctattcgac agcatgccaa tctcttcata aatcttttct
     7141 caatgatgct tggctctgga atgccagaac tacaatcttt tgatgacatt gcatacattc
     7201 gaaagaccct agccttagat aaaactgagc aagaggcttt ggagtatttc atgaaacaaa
     7261 tgaatgatgc acatcatggt ggctggacaa caaaaatgga ttggatcttc cacacaatta
     7321 aacagcatgc attgaactga agcttgtcga gaagtactag aggatcataa tcagccatac
     7381 cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa
     7441 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa
     7501 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     7561 tggtttgtcc aaactcatca atgtatctta tcatgtctgg atctgatcac tgatatcgcc
     7621 taggagatcc gaaccagata agtgaaatct agttccaaac tattttgtca tttttaattt
     7681 tcgtattagc ttacgacgct acacccagtt cccatctatt ttgtcactct tccctaaata
     7741 atccttaaaa actccatttc cacccctccc agttcccaac tattttgtcc gcccacagcg
     7801 gggcattttt cttcctgtta tgtttttaat caaacatcct gccaactcca tgtgacaaac
     7861 cgtcatcttc ggctactttt tctctgtcac agaatgaaaa tttttctgtc atctcttcgt
     7921 tattaatgtt tgtaattgac tgaatatcaa cgcttatttg cagcctgaat ggcgaatgg
//
