LOCUS       sequence-137193-        6783 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 6783)
  AUTHORS   Johnson SM, Grosshans H, Shingara J, Byrom M, Jarvis R, Cheng A, 
            Labourier E, Reinert KL, Brown D, Slack FJ
  TITLE     RAS is regulated by the let-7 microRNA family.
  JOURNAL   Cell. 2005 Mar 11. 120(5):635-47.
  PUBMED    15766527
REFERENCE   2  (bases 1 to 6783)
  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: "Cell. 2005 
            Mar 11. 120(5):635-47."
COMMENT     Sequence Label: pGL3-KRAS
FEATURES             Location/Qualifiers
     source          1..6783
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(142..162)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    484..605
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(521..540)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     575..594
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(2173..3825)
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     3741..3759
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     regulatory      3822..3831
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3861..4057)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3875..4010
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     misc_feature    4112..4203
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     complement(4144..4163)
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     polyA_signal    4217..4265
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(4396..4851)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4533..4554)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4745..4764
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4878..4982
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4983..5051,5052..5843)
                     /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(5201..5220)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6014..6602
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6503..6522
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6756..6773
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgctgccg gcacctgtcc tacgagttgc atgataaaga agacagtcat aagtgcggcg
       61 acgatagtca tgccccgcgc ccaccggaag gagctgactg ggttgaaggc tctcaagggc
      121 atcggtcgac ggatccgctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
      181 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
      241 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
      301 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
      361 ctccgctcca tcgttcagat ccttatcgat tttaccacat ttgtagaggt tttacttgct
      421 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt
      481 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
      541 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
      601 tcttatcatg tctgctcgaa gcggccggcc gccccgactc tagcgttaaa ccattcaaag
      661 ttcacataaa ggtaattacc actaccttaa aaaaatgccc atctacatca aaaattcaag
      721 aggcctaaat atcccctcat aagcactgca gttcctgaag tatggccatt tctttctctg
      781 tgtagaaacg aggtactgtg taagtcttaa cgccctacct aaacagtgtt ccaaagatct
      841 gtagtttcac atagcaattc agaaatcata gtgattttta catagaagtt tccttgtctg
      901 tgaactagtt caggcacctg tttatttgta cccagataaa actattaatt tttaagtata
      961 ttttaattac ttatgcagag aaaactggaa tattacacat ttgggtcaat atgaatatct
     1021 gacatacacc ttaatgtgta cagtaattgt cccaaaagaa tcacagttat gccaaataaa
     1081 aaaacaatat aatcaagttt attcctttaa aacaatgaag tgatttacat aaagtagctt
     1141 gatcgaagag tttcagtgga atcgtagcaa aacaattata gagctggcac agagaccaaa
     1201 ccccttcttt gcaaaactaa aatacgcatc gtgttatctc tgggtcgtat accaaaggcc
     1261 ttagtaagat attacagacc acactagcac tacctaagga ccgggattat gtctcttgtt
     1321 tggggatacc atatacccag tgccttgtgc ggtgactggc atctggtagg cactcaataa
     1381 atatttgctg aataaatgag ttctacaaaa caggtttatg aggccaaggt gggtgaatca
     1441 cttgaggtca ggagttcgag accagcctgg ccaacagggt gaaaccccgt ctctcctaaa
     1501 aatacaaaaa ttagttgagt gtagtggcac acgcctgtaa tcccagctac tcaggaggcc
     1561 gaggcacgag aatcgcttga acctgggaga tggaggttgc agtgagctga gatggcgcca
     1621 ctgcattcca gcctgggtga caagagcgag actctgacac caaaaaaaaa aaagtaagct
     1681 tcatgaatta aagtattttt cattgcatga agatttctgg ttactaaaac aatggaatgt
     1741 attactgtta ccaggagtag tcctagttat agattaccta aggaattctt tcagcactat
     1801 ctttattaaa ttctccttcc actggatagg gttctgtcta ttcataccag gtttgaaaaa
     1861 tcctactgtc gctaatggat tgggcagcaa agagatgttc aaagcatcag ccaccacctg
     1921 aaaattagtg attaggtcaa atccctttat ggtatctgtc agattctctt gagccctgag
     1981 gaaataagat gtagggcatt tctgatgtga ctcagtggga aaacttcatg gagatatcca
     2041 cagcagcagt aaatcttatg gttaggggaa ttacaagtat taaaactgca tcaagtcatg
     2101 gggcatgtgg aaggtaggga ggcaagatga cactaatatg gaagaagagt cctaaaacga
     2161 gaatggctag aattacacgg cgatctttcc gcccttcttg gcctttatga ggatctctct
     2221 gatttttctt gcgtcgagtt ttccggtaag acctttcggt acttcgtcca caaacacaac
     2281 tcctccgcgc aactttttcg cggttgttac ttgactggcg acgtaatcca cgatctcttt
     2341 ttccgtcatc gtctttccgt gctccaaaac aacaacggcg gcgggaagtt caccggcgtc
     2401 atcgtcggga agacctgcga cacctgcgtc gaagatgttg gggtgttgga gcaagatgga
     2461 ttccaattca gcgggagcca cctgatagcc tttgtactta atcagagact tcaggcggtc
     2521 aacgatgaag aagtgttcgt cttcgtccca gtaagctatg tctccagaat gtagccatcc
     2581 atccttgtca atcaaggcgt tggtcgcttc cggattgttt acataaccgg acataatcat
     2641 aggacctctc acacacagtt cgcctctttg attaacgccc agcgttttcc cggtatccag
     2701 atccacaacc ttcgcttcaa aaaatggaac aactttaccg accgcgcccg gtttatcatc
     2761 cccctcgggt gtaatcagaa tagctgatgt agtctcagtg agcccatatc cttgcctgat
     2821 acctggcaga tggaacctct tggcaaccgc ttccccgact tccttagaga ggggagcgcc
     2881 accagaagca atttcgtgta aattagataa atcgtatttg tcaatcagag tgcttttggc
     2941 gaagaaggag aatagggttg gcaccagcag cgcactttga atcttgtaat cctgaaggct
     3001 cctcagaaac agctcttctt caaatctata cattaagacg actcgaaatc cacatatcaa
     3061 atatccgagt gtagtaaaca ttccaaaacc gtgatggaat ggaacaacac ttaaaatcgc
     3121 agtatccgga atgatttgat tgccaaaaat aggatctctg gcatgcgaga atctcacgca
     3181 ggcagttcta tgaggcagag cgacaccttt aggcagacca gtagatccag aggagttcat
     3241 gatcagtgca attgtcttgt ccctatcgaa ggactctggc acaaaatcgt attcattaaa
     3301 accgggaggt agatgagatg tgacgaacgt gtacatcgac tgaaatccct ggtaatccgt
     3361 tttagaatcc atgataataa ttttttggat gattgggagc tttttttgca cgttcaaaat
     3421 tttttgcaac ccctttttgg aaacgaacac cacggtaggc tgcgaaatgc ccatactgtt
     3481 gagcaattca cgttcattat aaatgtcgtt cgcgggcgca actgcaactc cgataaataa
     3541 cgcgcccaac accggcataa agaattgaag agagttttca ctgcatacga cgattctgtg
     3601 atttgtattc agcccatatc gtttcatagc ttctgccaac cgaacggaca tttcgaagta
     3661 ctcagcgtaa gtgatgtcca cctcgatatg tgcatctgta aaagcaattg ttccaggaac
     3721 cagggcgtat ctcttcatag ccttatgcag ttgctctcca gcggttccat cttccagcgg
     3781 atagaatggc gccgggcctt tctttatgtt tttggcgtct tccatggtgg ctttaccaac
     3841 agtaccggaa tgccaagctt tttgcaaaag cctaggcctc caaaaaagcc tcctcactac
     3901 ttctggaata gctcagaggc cgaggcggcc tcggcctctg cataaataaa aaaaattagt
     3961 cagcgatggg gcggagaatg ggcggaactg ggcggagtta ggggcgggat gggcggagtt
     4021 aggggcggga ctatggttgc tgactaattg agatgcagat cgcagatctc gagcccgggc
     4081 tagcacgcgt aagagctcgg tacctatcga tagagaaatg ttctggcacc tgcacttgca
     4141 ctggggacag cctattttgc tagtttgttt tgtttcgttt tgttttgatg gagagcgtat
     4201 gttagtacta tcgattcaca caaaaaacca acacacagat gtaatgaaaa taaagatatt
     4261 ttattgcggc cgctccaagt acctcccgta ccttaatatt acttacttat catggtagct
     4321 tgggctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
     4381 tgaatggcga atggcaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     4441 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     4501 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     4561 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     4621 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     4681 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     4741 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     4801 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca
     4861 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     4921 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     4981 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     5041 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     5101 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     5161 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     5221 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     5281 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     5341 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     5401 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     5461 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     5521 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     5581 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     5641 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     5701 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     5761 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     5821 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     5881 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     5941 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     6001 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     6061 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     6121 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     6181 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     6241 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     6301 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     6361 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     6421 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     6481 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     6541 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     6601 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     6661 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     6721 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     6781 aag
//
